Mapping of Italian
research excellence in
Neurodegenerative
Diseases
1. MECHANISMS OF NEURODEGENERATION
A.
Macromolecular interactions and Neurodegeneration
1. Public health. Therapeutic strategies for the
prevention and tratment of neurodegenerative
disease. Molecular basis of amyloid aggregation
2. Protein conformational transitions that trigger the
aggregation processes associated to
neurodegenerative diseases : a nanotechnological
approach
3. Myelin proteins and demyelinating deseases
4. Conformational transitions of proteins from native
to amyloid form
5. Role of altered composition of plasma membrane
complex lipids in lipid-protein interactions,
membrane organization and neurodegeneration
6. Alterations of proteolytic pathways in
neurodegenerative diseases"
7. Optical and biomolecular methods for the
functional investigation of neural circuits in vivo
8. Molecular mechanisms in neurotransmission:
effects in neuron and glia phenotypes in synaptic
efficiency and synapropathy
9. Presynaptic molecular machinery regulationg
glutamate release an pathophysiological
mechanism in amyotrophic lateral sclerosis
10. Toward exploiting the screening potential of
miRNome for the identification of genes and
pathways involved in neurodegenerative disorders
11. Identification of signaling pathways in
neurodegenerative proteinopathies
12. Neurotensin and neurotensin antagonists in an
animal model of Parkinson disease: therapeutic
perspectives and role of NMDA/neurotensin
interaction.
B.
Pier Luigi San Biagio
[email protected]
Bruno Samorì
[email protected]
Eugenia Polverini
[email protected]
Maria Grazia Bridelli
[email protected]
Sandro Sonnino
[email protected]
Angelo Poletti
[email protected]
Antonio Malgaroli
[email protected]
Gian Giacomo Consalez
[email protected]
Flavia Valtorta
[email protected]
Maurizio Popoli
[email protected]
Daniela Tardito
[email protected]
Maria Penuto
Maria [email protected]
Tanganelli Sergio
[email protected]
Neurotoxic stimuli
1. CSF Glycomics in AD and other neurodegenerative
disease
2. Intracellular signalling during neurodegenerative
events triggered upon APP overexpression
Domenico Garozzo
[email protected]
Maurizio Taglialatela
[email protected]
Claudio Russo
[email protected]
3. Effect of Age and neurodegenerative disorders in
central neurotrasmission: focus on neurotrasmitter
relase from neuron and glia
Mario Marchi
[email protected]
4. Developing competitive in “ vitro” and “vivo”
models to study Alzheimer’s disease
5. Evaluation of mesenchymal stell cells (MSCs) effect
in Alzheimer’s disease rat models competitive in “
vitro” and “vivo” studies
6. New Tratment strategies (Neurodegeneration and
Neuroprotection)
7. Pathways leading to tau pathology – Role of nuclear
tau in neurodegeneration in fronto-temporal
dementia
8. Cholesterol and amyloid-beta: is there a relation?
-
Patrizia Hrelia
[email protected]
Fabrizio Tagliavini
[email protected]
Roberta Ricciarelli
[email protected]
Specific type 4 phosphodiestrerase inhibitors and
their impact on the production of amyloid-beta
Cholesterol and amyloid-beta: is there a relation?
Role of non muscle myosin IIB in the processing of
the amyloid precursor protein (APP)
9. Basic research
- Genetic susceptibility to Alzheimer disease and
genome wide association studies
- Biobanking
10. Neurotoxic stimuli, synaptic dysfunction
11. Neuroprotection by chemokines
12. Phatogenesis and therapeutic targets for SBMA
13. Malattia di Alzheimer e trasduzione del segnale
legata al calcio
14. Mechanisms of neurodegeneration and
enhancement of adult neurogenesis to improve
cognitive functions in down syndrome
15. Pathological mechanisms of neurodegeneration
triggered by tau and beta-amyloid
16. Oxidative stress and neurodegeneration
C.
Francesca Ruberti
[email protected]
Mariarosaria Miloso
[email protected]
Dario Finazzi
[email protected]
Calro Sala
[email protected]
Cristina Limatola
[email protected]
Fabio Benfenati – Mara Pennuto – Chiara
Scaramuzzino
[email protected]
Armando Genazzani
[email protected]
Andrea Contestabile
[email protected]
Laura Gasparini
[email protected]
Michele Mazzanti
[email protected]
Neuro inflammation
1. Role of vascular inflammation and leukocyte
trafficking in neurodegenerative diseases
2. Regional vulmerability of the brain to
neurodegeneration and immune regulatory
mechanisms
3. Neurotoxic stimuli: role of astrocytes and microglia
4. Gender and hormonal/endocrine signals in
neuroinflammation role in alzheimer’s disease and
Gabriela Constantin
[email protected]
Marina Bentivoglio
[email protected]
Michela Matteoli
[email protected]
Elisabetta Vegeto
[email protected]
brain ageing
5. Role of pro-and anti- inflammatory cytokines in the
prognosis and progression of neurodegenerative
diseases
6. Study of the role of pro-inflammatory chemokines
in neuroinflammatory disorders and analysis of the
therapeutic potential of chemokine synthesis
inhibitors
7. Role of inflammation and microglial activation in
neurodegenerative diseases
8. Role of inflammation in Alzheimer’s disease: study
of pro-inflammatory cytokines for the identification
of new pathogenic and diagnostic targets
D.
Barbara Viviani
[email protected]
Claudio Milanese
[email protected]
Luisa Minghetti
[email protected]
Paola Bossù
[email protected]
Oxidative Stress and neurodegeneration
1. Dopamine and oxidative stress in the pathogenesis
of Parkinson's disease
2. Mitochondria, apoptosis and neurodegeneration:
characterization of mitochondrial dysfunction in
neurodegenerative diseases
- Characterization of the molecular mechanisms
underlying aging, cell senescence and
neurodegenerative diseases
3. Structural organization of the mitochondrial
respiratory chain, generation of reactive oxygen
species and neurodegeneration
4. Post-genomic biochemestry of neurodegenerative
diseases
Marco Bisaglia
[email protected]
Antonella Bobba
[email protected]
Giorgio Lenaz
[email protected]
Annalisa Santucci
[email protected]
Trophic factors involvement in the neurodegeneration
process
E.
F.
Endocrine and Metabolic Factors
1. miRNA as sensors of neuronal stress and
modulators of synaptic plasticità and memory
deficits
2. Developing competitive animal models to study
Alzheimer’s disease
Paola Fragapane
[email protected]
3. Genetic susceptibility to Alzheimer’s disease (AD)
in women: AD as a gender disease
4. Genetic susceptibility to Alzheimer’s disease
(Epigenetic maks of susceptibility to Alzheimer
disese)
5. Role of altered lipid and cholesterol metabolism in
neurodegenerations
Rosa Maria Corbo
[email protected]
Lucia Migliore
[email protected]
Roberto Rimondini
[email protected]
Laura Colombaioni
[email protected]
6. Role of neuroactive steroids in the
neurodegeneration process
7. Selective Alzheimer Disease Indicator -1 (Seladinò1): a liker between cholesterol, oxidative stress
and Alzheimer’s disease
8. Nuclear receptors, coregulators, neuroactive
steroids and epigenetics in neurodegenerative
disease associated to metabolic disorders
G.
4. Synaptic deficits in neurodegenerative disease
Ubaldo Armato
[email protected]
Manuela Marcoli
[email protected]
Luciano Domenici
[email protected]
Evelina Chieregatti
[email protected]
Intracellular Traffiking
1. Membrane traffic and neurodegenerative diseases:
role of Rab proteins and their effectors
2. Basic research. Neurodegenerative diseases
I.
Donatella Caruso
[email protected]
Cell interactions
1. The Astrocyte, not just a Bystander in Alzheimer’s
disease
2. Synapse and Amyloid-beta: effects on astrocyte
processes, nerve terminals and network synaptic
activity
3. Alzheimer’s disease, Ageing
H.
Roberto Cosimo Melcangi
[email protected]
Roberto Maggi
[email protected]
Cecilia Bucci
[email protected]
Paolo Macchi
[email protected]
Animal models
1. Developing competitive animal models to study
Alzheimer’s disease
2. Synaptic dysfunction in neurodegeneration:
characterization of gene product in functional
synaptic networks under physiological and
patological conditions
3. Study of early onset Alzheimer disease in a model
for Down syndrome, the Ts65Dn mouse
4. Iron and Neurodegeneration
5. Non neurotoxic activity of Beta amyloid in animal
models
6. Neurochemical and morphological in vivo and in
vitro models in neurodegenerative diseases.
Adalberto Merighi
[email protected]
Monica DI Luca
[email protected]
Renata Bartesaghi
[email protected]
Sonia Levi
[email protected]
Stefano Govoni
[email protected]
Tiziana Antonelli
[email protected]
2. PHARMACOLOGY
Drug Design
A.
1. Elementary mechanisms of neurodegenerative
diseases
2. Innovative drug discovery strategies for Alzheimer’s
disease
3. Development of ligands specifically targeting the
Translocator Protein 18 kDa in Alzheimer disease
4. Structural studies of acetylcholinesterases and their
inibitors: implicantions for the design of new antialzheimer drugs
Nino Russo
[email protected]
Carlo Melchiorre
[email protected]
Federico Da Settimo
[email protected]
Alberto Cassetta
[email protected]
Drug screening
B.
1. Developemt of competitive animal models for ADGLP grade animal facility and behavioural testing
Laura Calzà
[email protected]
2. Yeast model to seek for molecules which can
reduce mitochondrial mutability
Iliana Ferrero Fortunati
[email protected]
3. Innovative technologies for the therapy of the
neurodegenerative diseases.
Luca Ferraro
[email protected]
Drug Delivery
C.
1. Nanotechnology for health: application in the brain
diseases
2. Hybrid drug containing reservoirs for long term
release in brain diseases
3. New nanodevices targeting beta-amyloid and
overcoming the blood-brain barrier for the therapy
of Alzheimer disease
MA Vandelli
Giovanni Tosi
[email protected]
Angelo Montenero
[email protected]
Massimo Masserini
[email protected]
Novel Drugs
D.
1.
2. Synthesis and screening of new AChE/BuChE
inhibitors
3. Drug design and synthesis of therapeutic agent for
neurodegenerative disorders, particularly for
Alzheimer disease
4. Development of a novel therapeutic strategy for AD
based on a natural variant of amyloid beta that
Lamberto Maffei
[email protected]
Aldo Andreani
[email protected]
Olga Bruno
[email protected]
Fabrizio Tagliavini
[email protected]
hinders amyloidogenesis
5. Second generation recombinant Abeta peptide
immunogens as prototype vaccines for AD
treatment
6. Pediatric neurodegenerative diseases
7. Ocular NGF administration as a novel non invasive
approach to protect brain-NGF target neurons that
degenerative in Alzheimer’s disease
8. New symptomatic and neuroprotective terapie for
parkinsons’s disease
9. Innovative neuro-reparative strategies via the
implementation of endogenous neurogenesis and
gliogenesis: focus on the new P2Y-like GPR17
receptor
10. Pharmacological modulation of adult neurogenesis
11. Study of the relationship between the
structure/morphology of amyloid beta peptide
aggregates and their toxicity in cellular, neuronal
and animal models"
E.
Simone Ottonello
[email protected]
Maurizio Scarpa
[email protected]
Paola Tirassa
[email protected]
Micaela Morelli
[email protected]
Maria Pia Abbracchio
[email protected]
Maria Grazia Grilli
[email protected]
Fabrizio Chiti
[email protected]
Non-Pharmacological Therapies
1. Psychoeducational intervention for patients and
care givers in ParKinson’s Disease and other
neurodegenerative diseases.
2. Physical excercise and cognitive decline
3. New tratment strategies (non pharmacologic
therapeutic strategies in neurodegenerative
diseases)
4. Non invasive brain stimulation an integrated
approach to neurorehabilitation in Alzheimer
disease
5. Early cognitive and electroencephalographic
markers of normal and pathological ageing
6. Transplantation of iPS-derived dopaminergic
neurons for cell replacement in Parkinson's disease
7. Neural re generation in the cerebellum:
development of cell replacement strategies for the
management of spinocerebellar ataxias
8. Physical activity programs for elderly subjects with
different grades of cognitive impairment
9. Promotion of Adapted Motor Activity among
patients with Parkinson’s disease
Pio Enrico Ricci Bitti
[email protected]
Maurizio Taglialatela
[email protected]
Alfonso Di Costanzo
[email protected]
Andrea Stracciari
[email protected]
Orazio Zanetti
[email protected]
Patrizia Bisiacchi
[email protected]
Vania Broccoli
[email protected]
Gian Giacomo Consalez
[email protected]
Ferdinando Rossi
[email protected]
Federico Schena
[email protected]
Enrico Granieri
[email protected]
[email protected]
[email protected]
10. New treatment strategies in Neurodegenerative
diseases with Deep brain Stimulation
F.
Drug Pharmacogenetics
1. Role of pharmacogenetics to identify the
personalized treatment of dementia and
depression.
G.
Mariachiara Sensi
[email protected]
Alberto Pilotto
[email protected]
Clinical studies
1. Neurocognition in Alzheimer Disease
Carlo Caltagirone
[email protected]
3. HUMAN/CLINICAL RESEARCH
A.
Human –epidemiology/risk factors
1. Pathogenic mechanisms, early diagnosis and
prevention of neurodegenerative diseases
following pesticide intake in animal model
2. Effects of traffic-related particulate matter on
transcription of genes implicated in
neurodegeneration
3. Cronobiology and neurodegeneration
4. Gene-metal interaction as determinant of
neurodegenerative diseases
5. Using prevention to reduce the burden of
Alzheimer’s disease
6. Construction and development of "Alzheimer-WebGIS".
Geographical correlation between Alzheimer's
disease and environmental parameters. Method for
mapping population-based case-control studies.
7. Prospective evaluation of biomarkers of
inflammation in Mild Cognitive Impairment
subtypes, and their role in the conversion from
cognitive impairment to dementia and Alzheimer's
disease
8. Epidemiology and risk factors for Mild Cognitive
Impairment, Alzheimer's disease and dementia.
Population-based 7-year follow-up study.
9. [moved to B.5 Epidemiology genetics]
10. Preclinical Diagnosis
11. Interactions between the products of the Herpes
Gabbianelli Rosita – Cinzia Nasuti
[email protected]
Micaela Caserta
[email protected]
Valerio Carelli
[email protected]
[email protected]
Roberto Lucchini
[email protected]
Stefano Mattioli
[email protected]
Giuseppe Mele
[email protected]
Calogero Caruso
[email protected]
Roberto Monastero
[email protected]
Emilio Di Maria
[email protected]
Carlo Caltagirone
[email protected]
Manservigi Roberto
simplex genome and Alzheimer's disease
susceptibility genes
B.
Epidemiology genetics
1. Genetic susceptibility to Alzheimer’s disease and
genome wide association studies
2. Genetic susceptibility to Alzheimer’s disease:
biobanking
C.
[email protected]
Luisa Benussi
[email protected]
Maria Del Zompo
[email protected]
Biomarkers and early diagnosis and imaging
1. Plasma biomarkers of oxidative stress and
inflammation in early detection and follow-up of
dementia.
2. Vitamins and antioxidants in aging and
neurodegenerative diseases.
3. Phenotypic and genotypic markers of early
Alzheimer’s disease
4. Prioritization and annotation of AD genes, their
variants and their interaction networks
5. Cerebrospinal fluid cytokine and chemokine levels
as potential biomarkers for early diagnosis of
Alzheimer’s disease
6. Individual evaluation of cognitive decline: definition
of low cost non- invasive examination battery and
implementation of a predictive algorithm to
identify Mild cognitive impairment
7. Impiego dei markers biologici e neuro radiologici
nella diagnosi differenziale precoce
8. Biomarkers in Mild cognitive Impairment: genetics
and biochemical and neuroimaging studies for
early diagnosis.
Patrizia Mecocci
[email protected]
9. A multimodality index for early diagnosis of
Alzheimer’s disease
10. Lexical semantic skills, APOE and neuroanatomical
substrate of semantic memory in early diagnosis of
Alzheimer's disease
11. Cognitive neuroscience of dementia: clinical and
theoretical aspect
12. Mapping structural and functional brain network
damage in neurodegenerative diseases
13. Studies are aimed at investigating whit PET the
brain functional parameters and neurotransmission
systems in neurological and psychiatric diseases
14. Global proteomics of human fibroblast for the
identification of early biomarkers for dementia and
Alzheimer’s disease
15. Parkinson disease and parkinsonisms
Flavio Nobili
[email protected]
Paolo Caffarra
[email protected]
16. Mild cognitive impairment
Salvatore Monaco
[email protected]
Rita Casadio
[email protected]
Elio Scarpini
[email protected]
Paolo Maria Rossini
[email protected]
Orazio Zanetti
[email protected]
Sandro Sorbi
[email protected]
Stefano F. Cappa
[email protected]
Massimo Filippi
[email protected]
Daniela Perani
[email protected]
Maurizio Popoli
[email protected]
Gianfranco Spalletta
[email protected]
Gianfranco Spalletta
17. Electrophysiological study of striatal physiology in
normal conditions and experimental parkinsonism:
use of genetic and pathogenetic animal models
18. Celebrospinal fluid biomarkers in alzheimer’s
disease and other neurodegenerative disease
D.
Lucilla Parnetti
[email protected]
Diagnosis: biomarkers
1. Proteomics of cognitive and movement disorders:
Identification of molecular mechanisms associated
with disease onset and phenotypic variability of
frontotemporal lobar degeneration
2. Peripheral markers of oxidative stress,
excitotoxicity and abnormal protein metabolism in
neurodegenerative disorders.
3. Validation and search of CSF biomarkers for the
early differential diagnosis of cognitive impairment.
Development of new diagnostic approaches for
prion diseases in humans.
4. Analisi Biochimica di parametri di stress ossidativo
5. Cognitive decline in multiple sclerosis
6. Search of predictive markers of AD based on
amplification of disease-specific amyloid-beta
oligomers
7. Mild cognitive impairment and dementias
E.
[email protected]
Paolo Calabresi
[email protected]
Roberta Ghidoni
[email protected]
Carlo Ferrarese
[email protected]
Piero Parchi
[email protected]
Gabriele Siciliano
[email protected]
Laura Calzà
[email protected]
Fabrizio Tagliavini
[email protected]
Gianfranco Spalletta
[email protected]
Diagnosis: neuropsychology
1. Handwriting analysis as a novel diagnostic tool for
Alzheimer’s disease in the clinical and forensic
settings
2. Amnesic Mild Cognitive Impairment:
neuropsychological features and new diagnostic
criteria (Dubois et Al., 2007)
3. Predictors of conversion from MCI to dementia
Maurizio Balestrino
[email protected]
Angiola Maria Fasanaro
[email protected]
Roberto Gallassi
[email protected]
4. Individual evaluation of cognitive decline; validation Ildebrando Appollonio
of ultrafast screening tools.
[email protected]
F.
Biobanking Bioinformatics
G.
Ambient assisted living and socioeconomics
1. Home-based Empowered Living for people with
dementia and Parkinson’s disease
Roberto Monastero
[email protected]
2. Neuro-rehabilitation and brain functional imaging:
rehabilitation customization driven by best
exploitation of celebral plasticity and health system
sustability.
3. Role of information and communication technology
(ICT) systems to improve quality of life, quality of
care and safety of older patients with cognitive
decline
4. A transdisciplinary orented proposal from a social
sciences perspective
Alessandra Pedrocchi
[email protected]
Alberto Pilotto
[email protected]
Vincenzo Maria Bruno Giorgino
[email protected]
1. MECHANISMS OF NEURODEGENERATION
A-
Macromolecular interactions and
Neurodegeneration
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Dr. Pier Luigi San Biagio
e-mail: [email protected]
phone: +39 091 6809311
cell: +39 329 410 5564
Istituto di Biofisica - CNR, Via Ugo La Malfa, 153 , 90146 Palermo (Italy)
Many untreatable neurodegenerative human disorders are associated with the
aggregation of misfolded or natively unfolded proteins into amyloid fibrils. Our
research activity concerns Alzheimer’s Parkinson’s and Albers-Schönberg diseases
with the shared perspective of relating the understanding of the pathway leading
to the fibrils formation to the design of appropriate drugs capable on preventing
the protein aggregation.
Alzheimer’s disease. Recent results suggested that the main neurotoxic species in
Alzheimer’s Disease (AD) would not be the
insoluble protein aggregates, made of extracellular deposits of Beta-amyloid
peptide (Ab) fibrils, but rather the soluble oligomeric
species, including small globular structures, 2.7 to 6.0 nm in diameter, and
“protofibrils”. Conformational constrains imposed by small
organic molecules could play a key role in modulating the fibrillogenesis process,
so providing effective therapeutic tools to target both
oligomeric and fibrillar species. In this perspective, polycyclic aromatic molecules
could be of special interest as they might disrupt the
molecular architectures precursors of fibrils by means of aromatic interactions,
like stacking interactions with tyrosine and
phenylalanine residues of Ab. In particular, natural polyphenols, composed of one
or more aromatic phenolic rings, are a class of
phytochemicals found in high concentrations in wine, tea, nuts, berries, fruits,
cocoa, and an ample assortment of other plants. Some of
them have been demonstrated to have anti-oxidant and anti-amyloidogenic
effect and it has been speculated that they could prevent
the development of Alzheimer's disease, not only through scavenging of reactive
oxygen species, but also through directly inhibiting
the deposition of fibrillar beta-amyloid in the brain. In the present project, we’ll
test some natural molecules, such as ferulic acid (FA), a
phenolic anti-oxidant present in fruit cell walls, hypericin (Hyp), a natural pigment
extracted from Hypericum perforatum, widely used as
a mild antidepressant and epigallocatechin gallate (EGCG), one of the major
flavonoids of green tea. The aggregation process of Ab in
the presence of these natural molecules will be studied by light scattering,
steady-state and time-resolved fluorescence, FTIR and CD
techniques. The analysis of the correlations between the effects of the studied
compounds on the various stages of amyloid fibril
formation, and their known physicochemical properties, will provide novel
insights on the specific role of hydrophobic and aromatic
interactions. Encouraging results have already been obtained using hypericin
which has been shown to affect and to interfere with the
early stages of polymerization process. The furtherance of this type of study,
extended to other small molecules like FA and EGCG,
hopefully will meet the goal of designing effective therapeutic tools to ideally
target both oligomeric and fibrillar species, thus
contributing to devise AD prevention strategies.
Parkinson’s disease. Alpha-synuclein (aS) is the main constituent of Lewy bodies,
intraneuronal fibrillar inclusions which are present
in all patients affected by Parkinson’s disease (PD). From genetic and biochemical
studies it is believed that aS is involved in the
disease. AS is a natively unfolded protein with the ability to acquire secondary
structure upon interaction with membranes or with itself.
It is linked to PD by two evidences: the accumulation of amyloid fibrils of the
protein and the autosomal dominant forms of the disease
(A53T, A30P and E46K mutants). Both the biological role of this protein and the
mechanisms involved in the ethiopathogenesis of PD
are still unknown. The protein is located at the presynaptic terminal of neurons in
all the CNS, where it exists free in the citosol or
bound to synaptic vesicles. The membrane binding causes the formation of an
amphipatic alpha-helix in the first part of the protein, which lies at the membrane
surface without crossing the bilayer. Recent evidences show that aS is able to
bind and permeabilize cell
membrane. Our investigation will be devoted (i) to the characterization, at the
single molecule level, of the channel like ability of aS and its architecture. The
pore-forming ability of monomers and aggregates of different sizes will be
investigated. Secondly, the lipid specificity will be also investigated by means of
high resolution NMR and MS (both maldi-tof and esi-ms). The ability of aS to
provoke or protect unsaturated acyl chains of membrane lipids from oxidation
will also be investigated, by studying with MS the products of oxidation. As
possible therapeutic perspectives, the organic or peptidic inhibitors of proteinprotein interaction or able to clog the already formed pores will be investigated.
Furthermore, if indications of any lipid specificity will be obtained, we will
develop lipid formulates suitable for sequestering aS lowering its level in solution.
Albers-Schönberg disease. Mutations in the gene coding for the lysosomal Cl-/H+
exchanger CLC-7 lead to recessive osteopetrosis
and Albers-Schönberg disease. Apart from the osteopetrotic phenotype the
recessive disease is characterized by severe generalized
neurodegeneration. Also knock-out of the plasma membrane localized CLC-2 lead
to neurodegeneration (retinal degeneration and
leukoencephalopathy). The mechanisms underlying these phenotypes are
unclear. We are studying the functional properties of these
two proteins using electrophysiological and optical methods, combined with
mutagenesis, in order understand how their dysfunction
may lead to neurodegeneration. In particular, we consider these proteins as
potential risk factors for more generalized forms of
neurodegeneration.
Area di interesse
identificata
Finanziamenti ricevuti
Titolo progetto
Public health. Therapeutic strategies for the prevention and treatment of
neurodegenerative
diseases. Molecular basis of amyloid aggregation
Ente finanziatore
Progetto strategico di ricerca finalizzata sulla malattia di Alzheimer (Art. 12/bis D.Lgs
229/99),
“Beta-amyloid deposit on the cell membrane: role of metal ions and free radicals”
Italian 'Ministero della Salute”
Durata progetto
24 months, started 2001
Abstract del progetto
Ente finanziatore
Characterization of some key molecular mechanisms for the neuronal damage connected
to AD. Specific objectives: (i) to identify of the key steps responsible for the amyloid
depost formation on membrane model systems in vitro; (ii) to evaluate the role of the
membrane lipid composition on the deposit; (iii) to understand the role of metal ions and
oxidative stress on
the fibril growth both in solution and on membranes; (iv) to establish an in vitro test for
evaluating the ability of drugs to inhibit bA aggregation.
“Animal neuropathies: molecular and functional analysis of the prionic proteins in Sicilian
bovine race ”
Italian 'Ministero della Salute
Durata progetto
24 months, started 2003
Abstract del progetto
Titolo progetto
The main objective of the present proposal is to understand the mechanisms by which
wild type and mutant prion proteins become misfolded in such a way as to provoke
neurodegenerative diseases becoming transmissible agents. An extensive study of the
aggregation process will be done by using static and dynamic light scattering and
fluorescence techniques.
Prin 2005 project: “The effects of metal ions on the protein aggregation processes”
Ente finanziatore
Italian 'Ministero Università e Ricerca'
Durata progetto
24 months, started 2005
Abstract del progetto
Ente finanziatore
The project concerns the study of the effects of metal ions in solution on the protein
aggregation processes. Main goals are (i) to understand the molecular mechanisms
relevant for amyloid aggregation and to establish the relevance of metals in modulating
solvent mediated protein-protein interactions.
“Electrophysiological characterization of the alpha synuclein channels, a protein involved
in Parkinson’s disease”
Local Bank Foundation CARITRO
Durata progetto
24 months, started 2008
Abstract del progetto
Ente finanziatore
The aim of the project is: i) to characterize the channel like activity of alpha synuclein (aS)
and its role in the disease; ii) to distinguish the most active aS form; iii) to verify the ability
of aS to bind to membranes and lipid requirements; iv) to investigate the role of aS in the
prevention of the oxidation of unsaturated fatty acid chains
Prin 2008 project: “Inhibition of beta-amyloid peptide aggregation by some natural
compounds”
Italian 'Ministero Università e Ricerca
Durata progetto
24 months, started 2010
Abstract del progetto
Ente finanziatore
The aim of this project is to provide a molecular approach for preventing Abeta
aggregation and toxicity through the use of some natural molecules potentially able of
interact with Abeta by stabilizing or disrupt aggregates of different sizes.
“Membrane transport of chloride and protons: mechanism of function of CLC family
proteins involved in epilepsy and neurodegeneration”
Compagnia San Paolo
Durata progetto
36 months, started 2009
Abstract del progetto
CLC-2 is a plasma membrane localized Cl- channel whereas CLC-7 is a lysosomal Cl-/H+
exchanger. Knock-out of CLC-2 in mice leads to retinal degeneration and
leukoencephalopathy and knock-out of CLC-7 and human mutations in the CLCN7 gene
Titolo progetto
Titolo progetto
Titolo progetto
Titolo progetto
lead
to generalized neurodegeneration. The aim of the project is to understand better the
transport
function of these two proteins in order to get insight into their causative role in these
forms of
neurodegeneration.
Nome
Bruno Samorì
Contatti
E mail: [email protected]
Istituto/Dipartimento
Laboratory of Nanoscience and Nanobiotechnology, Department of
Biochemistry, University of Bologna (Italy)
(http://biocfarm.unibo.it/samori/)
Proposta di ricerca
Area di interesse identificata
Basic research
Abstract
According to an emerging view, the aggregation-prone proteins associated
to neurodegenerative, Parkinson’s, Alzheimer’s, and Creutzfeldt-Jakob
diseases, can switch back and forth between functional and
amyloidogenic/prionogenic conformations. Their conformational
polymorphism poses tremendous challenges to standard methods in
structural biology, which mask the complexity of their behaviour by
recording observables as time and ensemble averages. Single-molecule
methodologies can instead follow the time trajectories of individual
molecules or molecular assemblies, can map their energy landscapes, and
obtain distributions of molecular properties. These capabilities can provide
information about the monomeric state of an amyloidogenic protein and
about the very early stages of its amyloid cascade. This information is
crucially needed for the design of new drugs against Parkinson’s or
Alzheimer’s disease that must be capable to target upstream of the
formation of the most neurotoxic oligomeric species. In fact concerns are
now growing over the possibility that more harm than cure can be created
whenever the deposits in the central−nervous−system, are targeted
instead: in this way their dynamic equilibrium with the oligomeric forms
can be shifted back to the latter.
We have recently shown that Atomic Force Microscopy-based Single
Molecule Force Spectroscopy (AFM-SMFS) can single out the different
monomeric conformers of α-synuclein (the intrinsically unstructured
protein associated to Parkinson’s disease) and of its mutants associated to
genetic PD. (1,2). So far AFM-SMFS has been the only methodology that
made it possible to achieve this goal.
Ongoing projects:
a)
The integration of AFM-SMFS with Optical tweezers (OT) (in coll.
with Carlos Bustamante (UC Berkeley).
b) The determination of alpha synuclein aggregation step targeted
by different potential drugs or chaperons, by using an integrated
approach based on AFM- and OT-SMFS, AFM-imaging,
Fluorescence Polarization Spectroscopy ( in collaboration with
Luigi Bubacco, University of Padova)
c)
The conformational equilibria or prion proteins (in coll. with G.
Legname SISSA Trieste, and Motomasa Tanaka, Riken, Japan)
(1) Sandal, M., et al (2008). PLoS Biol 6, 999-1008.
(2) Brucale, et al. (2009). Chembiochem 10, 176-83.
Finanziamenti ricevuti
Titolo progetto
Nanomanipolazione di singole molecole delle proteine coinvolte in
malattie neurodegenerative
Ente finanziatore
FIRB Nanobiotecnologie per dispositivi e sensori innovativi (G.U.
29.07.2005 n. 175)
FIRB Metodologie e tecnologie innovative per la farmaceutica. CINECA
RBNE03PX83
Durata progetto
2005-2008; 2005-2010
Abstract del progetto
Sviluppo metodologie di nanomanipolazione di proteine intrinsecamente
non foldate o disordinate
Titolo progetto
Meccanismi Neurodegenerativi nelle Malattie da Prioni (coordin. Catia
Sorgato)
Ente finanziatore
PRIN 2009
Durata progetto
2010-2012
Nome
Dr. Eugenia Polverini
Contatti
[email protected]
Istituto/Dipartimento
Department of Physics – University of Parma
Proposta di ricerca
Our studies are - and will be - focused on myelin proteins, to the aim of clarify their structure to function relationship
and the mechanisms involved in demyelinating diseases, including multiple sclerosis. Two main proteins are under
investigation: the Myelin Basic Protein (MBP) and the P2 myelin protein.
MBP is a highly post-translationally modified structural component of central nervous system myelin, the
multilamellar membrane wrapped around nerve axons. MBP has a high net positive charge and its principal role
seems to be the adhesion of the cytosolic surfaces of the compact myelin sheath, whose structural integrity
determines the speed of transmission of action potentials along the axon. However, MBP seems to be a
multifunctional protein, since it also interacts with other cytoskeletal and signalling proteins, tethering some of them
to the oligodendrocyte plasma membrane. The three-dimensional structure of MBP in situ is still unknown, due to its
intrinsic flexibility and dependence of conformation on environment. Therefore a neighbourhood-dependent
structural charcterization of fragtments is attempted. In the classic 18.5 kDa MBP sequence, there is a highly
conserved fragment, constituting two regions with different putative functionality. The first region seems to
associate with the membrane, and comprises the primary immunodominant epitope in multiple sclerosis; the second
one is a proline-rich segment, that has been predicted to be a ligand for protein kinase SH3-domains. Due to the
functional relevance of this fragment of MBP, we’re trying a structural characterization - in the absence and in the
presence of protein’s post-translational modifications - by means of molecular dynamics simulations in membrane
and of spectroscopic techniques (in collaboration with the University of Guelph, Ontario, Canada).
P2 myelin protein is a basic peripheral nervous system protein localized to the cytoplasmic spaces in the myelin
lamellae. It was recognized as the factor inducing the Experimental Allergic Neuritis, an animal model for the study of
the Guillain-Barrè syndrome, a demyelinating disease of the PNS in humans. The interest for the putative
autoantigenic role of the P2 protein and the need to understand its function in the myelin sheath has led to the study
of its structural characteristics and behaviour, basing on the solved crystal structures. By investigating its interaction
with different kinds of lipids, on the one hand we want to clarify its transport role and on the other hand we want to
verify its putative involvement in the compactness of PNS myelin sheat. Different approaches are carrying out:
molecular docking simulations, spectroscopical investigations and, in collaboration with the University of Oulu,
Finland, crystal X-ray diffraction and small-angle X-ray scattering.
Area di interesse identificata
myelin proteins and demyelinating deseases
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
University local research funds (FIL)
Nome
Prof. ROBERTO DE RENZI - Dr. MARIA GRAZIA BRIDELLI
Contatti
[email protected]
Istituto/Dipartimento
Physics department
University of Parma
Parco Area delle Scienze, 7/A, Parma
ITALY
Proposta di ricerca
The goal of our research is threefold:
1) study of the nucleation rate and the growth rate of the amyloidogenic process by controlling and modulating the
solution parameters such as temperature, pH, solvent composition, and peptide concentration, not only influencing
the final fibril state but also the formation kinetics. The analysis of the aggregation process will be performed in
solution by employing CD and Fluorescence spectroscopies by following the change of fluorescence emission of small
hydrophobic molecules, called amyloid ligands, which do not bind to specific proteins but are selective for protein
aggregates with a cross β-sheet conformation. Dynamic Light Scattering measurements will allow to monitor size and
shapes of the fibrils during the aggregation process.
2) study the conformational state of proteins in the native and amyloid form at very low hydration level by
investigating the structural properties of the first hydration shell. It is well known that proteins are surrounded in the
cell by a hydration shell formed by interacting water molecules that form patches locally distributed in dependence
on the heterogeneous surface chemistry or occluded in the internal cavities. This bound water in the crowded
cytoplasm may not be enough to fully cover the protein surface with a monolayer aqueous sheath, moreover about
0.4 g of water/g of protein are enough to assure protein normal functionality. Two experimental techniques can be
applied to investigate this structured water to gain information on the geometry of the protein surface and
secondary structure. Thermally Stimulated Depolarization Currents (TSDC) technique, applied in the past to a variety
of natural and synthetic biopolymers has recently been applied to the study of lysozyme. It has allowed recognizing
the main features of the hydration as a function of secondary structure, revealing a clear dominance of the βstructure in the amyloid form. Fourier Transform Infrared Spectrometry as well has proven to be highly versatile in
the analysis of protein conformation by monitoring the alteration in protein secondary structure as changes in the
Amide bands.
3) by means of the molecular dynamics simulation techniques, study the conformational changes that proteins
undergo under the critical environmental conditions that in vitro cause the fibrils formation. This approach could be
useful to investigate the pathway(s) of the transition and its molecular features.
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Conformational transitions of proteins from native to amyloid form.
University local research funds FIL 2008
Hydration structure analysis of Lysozyme amyloid fibrils
Ministero dell’Istruzione, dell’Università e della Ricerca
2y
Combined Thermally stimulated depolarization currents (TSDC) and FTIR
techniques have been employed to investigate the conformation of hen egg
white lysozyme in native and amyloid form, in the state of powder at very low
hydration level. The spectra reveal specific features that can be attributed to
water texture around the secondary structure adopted by the macromolecule.
Nome
Contatti
Tel.
E mail
Dipartimento
Proposta di ricerca
Sandro Sonnino
Via fratelli Cervi 93, 20090 Segrate (Mi) Italy,
+39 0250330360, fax +39 0250330365,
[email protected]
Medical Chemistry, Biochemistry and Biotechnology, University of Milan
Role of altered composition of plasma membrane complex lipids in lipid-protein interactions, membrane
organization and neurodegeneration.
Area di interesse identificata
1. MECHANISMS OF NEURODEGENERATION
F. Endocrine and Metabolic Factors
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Angelo Poletti
02-5031.8215
[email protected]
Dipartimento di Endocrinologia, Fisiopatologia e Biologia Applicata , Centro di
Eccellenza sulle Malattie Neurodegenerative Universita' degli Studi di Milano
via Balzaretti 9 - 20133 Milano
Macromolecular interactions and Neurodegeneration
Ente finanziatore
ROLE OF ALTERED PROTEOLYSIS IN THE DEGENERATION OF MOTOR NEURONS IN
in vivo AND in vitro MODELS OF FAMILIAL AMYOTROPHIC LATERAL SCLEROSIS.
INVOLVEMENT OF CONSTITUTIVE AND IMMUNO PROTEASOMES AND OF THE
AUTOPHAGY−LYSOSOME PATHWAY
TELETHON – Italy
Durata progetto
three years (end 2010)
Abstract del progetto
Amyotrophic lateral sclerosis (ALS) is a neurological disorder primarily affecting
motor neurons, i.e. the neurons responsible for the contraction of muscles that
control voluntary movements, speach, swallowing, respiration. The disease,
whose causes are still largely unknown, is characterized by a progressive
paralysis that leads to death for respiratory failure. As in some familiar forms of
ALS there are mutations in the gene
that codes for the enzyme superoxide dismutase 1 (SOD1), it has been possible
to create experimental models of the disease to facilitate the research on the
causes that lead to motor neurons death. Both in ALS patients and in the
experimental models the vulnerable neurons contain aggregates of abnormal
proteins, whose accumulation can be responsible for the degeneration. It is
therefore possible to hypothesize that a role in the pathology is played by
dysfunctions of the proteolytic mechanisms, i.e. of the mechanisms that normally
degrade intracellular protein. We propose to investigate whether motor neurons
degeneration in ALS is due to a failure of protein degradation, using two
experimental models (transgenic mice and a motor neuron cell line, both
expressing mutant human SOD1) and analyzing two different intracellular
proteolytic systems: the proteasomes and the autophagosome−lysosome
Motorneuron degeneration in Spinal and Bulbar Muscular Atrophy: from the
molecular mechanisms to the potential therapeutical approaches
TELETHON - ITALY
Three Years (End 2011)
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
CAG/polyglutamine (polyQ) diseases, the largest class of hereditary
neurodegenerative diseases, include SpinoBulbar Muscular Atrophy (SBMA), a
motorneuron disease linked to the androgen receptor (AR) mutation. SBMA is a
valuable model of polyQ diseases, because ARpolyQ neurotoxicity is triggered by
testosterone, the endogenous AR ligand, possibly via protein conformational
changes. In SBMA animal models, the symptoms are reversed by testosterone
removal suggesting that SBMA patients can be treated with drugs preventing
testosterone action. The overall objectives of the study are to find the
mechanism(s) by which ARpolyQ neurotoxicity can be modulated by
testosterone with the aim to identify novel therapeutical approaches for SBMA
and possibly indications useful for other polyQ diseases. Therefore, all the
studies proposed will be carried out in absence or in presence of testosterone,
and four different specific aspects, thought to be involved in SBMA, have been
selected:
− interactions of the two major intracellular degradative systems: the
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
ubiquitin−proteasome and the autophago−lysosome pathways in the removal of
the neurotoxic ARpolyQ; analysis of ARpolyQ solubility and aggregation, and their
modulation by selective AR modulators (SARM) and HSP−modulators.
− analysis of the axonal antero−retrograde transport in presence of ARpolyQ
activated by testosterone (and SARMs). This aim will include the expression
analysis of some proteins related to axonal outgrowth and controlled by
testosterone (neuritin or CGP15).
− analysis of the possible toxicity of ARpolyQ in muscle cells, and the interaction
between motorneuron and muscle cells in SBMA.
− analysis of the AR promoter activity to identified factors downregulating the
ARpolyQ levels in motor neuron. These studies will provide information of the
mechanisms by which ARpolyQ becomes neurotoxic after testosterone
interaction, that may be used to design novel therapeutical approaches for SBMA
and related diseases
Alterations of Axonal Functions and Neurodegeneration in Motor Neuron
Diseases
FONDAZIONE CARIPLO
Three Years (end 2012)
Motor neuron diseases (MNDs) are a group of sporadic or hereditary
neurodegenerative diseases characterized by selective motor neuron death. They
may affect children and adult patients with relentless progression rate leading to
amyotrophic paralysis and death. Several genetic, molecular and cellular
alterations have been reported in the different MNDs and interpreted as
potentially relevant in the pathogenesis responsible for neuronal death.
However, it is still unclear whether these alterations are cause or effects of initial
insults capable to perturb cell functions. The clinical trials attempted with the
aim to counteract these alterations have proven to be largely ineffective. A
common aspect of most MNDs is the alteration of motor neuron axons, thus
suggesting specific axonal function impairment. Axonal processes can reach a
considerable length (up to 1 meter) and are fundamental for motor neuron
functions and survival. Axons allow neuronal control of muscle contraction, but
also collect survival/trophic factors released from the muscle, and required for
motor neuron physiology. Nerve damages and axonal loss result in motor neuron
death and muscle atrophy. The project will focus on three major human MNDs,
spinal muscular atrophy (SMA), spino-bulbar muscular atrophy (SBMA) and
amyotrophic lateral sclerosis (ALS), affecting similar motor neuron population
with different rate of cell death. It will be based on advanced analytical
techniques and specific expertise of five research units. The units will utilize
complementary genomic, biochemical, molecular and cellular assays to study the
mechanisms of axonal dysfunctions in the MNDs studied. Since gene mutations,
gene expression impairment, and altered post-translational modifications of
specific proteins are involved in these MNDs, the project will be dedicated to
unravel such alterations at the genomic and molecular level with the final aim of
identifying novel common therapeutical targets. Main objectives: i)
development and/or improvement of motor neuron models of SMA, SBMA and
ALS; ii) analysis at the (nuclear and mitochondrial) genomic level of alterations
determined by the lack or the expression of proteins (FL-SMN/a-SMN, mutant
ARpolyQ, mutant SOD1) responsible for the MNDs selected; iii) morphological,
cellular, molecular and biochemical analyses of the effects of the considered
proteins on axonal functions/dysfunctions in the MNDs selected.Five different
research institutions (University of Milan, Center of Excellence on
Neurodegenerative Diseases; IRCCS Foundation Neurological Institute C. Besta;
IRCCS Italian Auxologic Institute; Mario Negri Institute for Pharmacological
Research; National Institute of Molecular Genetics) with specific expertise in the
basic mechanisms of neurodegeneration will be involved. The approval of this
project would enable to establish a critical mass of researchers devoted to MNDs
in Milan, allowing the recruitment and formation of young scientists.
Nome
Contatti
Istituto/Dipartimento
Antonio Malgaroli, Gian Giacomo Consalez
02 2643 4886
[email protected], [email protected]
Università Vita-Salute San Raffaele
Proposta di ricerca
Optical And Biomolecular Methods For The Functional Investigation Of Neural Circuits In Vivo.
Area di interesse identificata
Analisi della funzione sinaptica nei tessuti nervosi normali e in modelli di
neurodegenerazione
Finanziamenti ricevuti
Titolo progetto
Optical And Biomolecular Methods For The Functional Investigation Of Neural
Circuits In Vivo.
Ente finanziatore
Fondazione CARIPLO (pending)
Durata progetto
2 anni
Every human behavior depends on an activity change in one or more brain areas,
often with the contribution of very small subgroups of cells and synapses.
Conversely, when a brain disease whatsoever, a tumor, a vascular disorder, a
grey or white matter pathology, etc., modifies some behavioral aspect, inevitably
this effect arises from a change in neural activity. Hence, viewing the fine
structure of the brain by the most advanced imaging techniques is certainly
important but it would be more important to find some effective way to detect
changes in neuronal and synaptic activity during both physiological and
pathological processes. Indeed, for many different body organs, the kidney, the
heart, the liver, the pancreas, etc, the early detection of signs and symptomsand
the choice of an optimized therapeutic approach often depend more on the
availability of a large set of chemical and molecular markers of cell function
rather than on our ability to visualize fine details of the tissue anatomy.
Unfortunately techniques available today to monitor brain function, such as
E.E.G., fMRI, fNIRS, etc., either sample brain activity very indirectly, through
changes in metabolism and blood flow, or do not have the resolution required to
pinpoint changes in small networks of neuronal cells and synapses. At present,
anomalous states of brain activity can be detected only when they arise from
the concerted activity of large numbers of neurons and synapses, such as in
epilepsy, or when cell and/or synaptic and/or axonal damage becomes very
profound. For all the above reasons, the demand for a methodology that could
yield a more direct readout of in vivo neuronal and synaptic activity is extremely
high.
In this research project we are planning to further develop a methodology that
was recently designed in one of our labs to provide a very direct and sensitive
readout of brain activity. This is based on a family of recombinant molecules
which were designed in our labs to detect synaptic activaton via the application
of exogenous markers. This methodology is capable of revealing the physiological
activation of brain synaptic networks with very high sensitivity and resolution.
These striking results have prompted the generation of a series of transgenic
animal models and analytical tools that will be used for further in vivo validation
and for the selective probing of specific subgroups of neuronal cells and
pathways, both in physiological and pathological conditions. The final goal of this
project is to test the applicability of this approach to the quantitative analysis of
brain activity. We expect that this work will generate important information for
the development of similar approaches to the human brain.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Flavia Valtorta
Flavia Valtorta, Fabio Grohovaz, Daniele Zacchetti
02-2643-4826 (Valtorta) 4811 (Grohovaz), 4817 (Zacchetti)
[email protected], [email protected], [email protected]
San Raffaele Scientific Institute, Division of Neuroscience
Via Olgettina 58
20132 Milano, ITALT
Proposta di ricerca.
Molecular mechanisms in neurotrasmission: effects of neuron and glia phenotypes in synaptic efficiency and
synaptopathy
Comprehensive goal of our research is the elucidation of the molecular mechanisms that underlie communications
between cells of the brain, both in the adult and during development. This proposal involves the complementary
expertise and the tight collaboration of two research units: the NEUROPSYCHOPHARMACOLOGY UNIT (Flavia
Valtorta) and the CELLULAR NEUROPHYSIOLOGY UNIT (Fabio Grohovaz and Daniele Zacchetti).
The main form of communication between neurons is achieved at the synaptic level through regulated secretion of
neurotransmitter. This process is of paramount importance for information processing in the central nervous system
and is known to undergo dysfunction in a number of pathophysiological states. Substantial advancements have been
made concerning the identification of the proteins involved in the basic mechanism of neurotransmitter release.
Much less is known about the molecules that participate to the assembly of the complex synaptic machinery during
development.
Research in Valtorta’s group will develop along two main lines: Aim 1: Diseases of synaptic origin. The main focus will
be on the family of the synapsins and their link to epilepsy and autism. Synapsins are a family of brain
phosphoproteins involved in neurotransmitter release through regulation of synaptic vesicle availability. Recent data
from the lab show that synapsins have also an important function in the organization of synaptic contacts. Synapsin
KO mice develop an epileptic phenotype, and mutations in the human synapsin genes have been recently associated
with epilepsy and autism spectrum disorders. The central interest is to investigate the molecular roles of synapsin in
synapse formation and function, and how these roles relate to the development of brain pathology. Another project
concerns the identification of novel molecular interactors of synuclein and of their role in neurodegeneration
associated with Parkinson disease.
Aim 2: Membrane trafficking in neuronal development. The process that leads from undifferentiated, round neuronal
precursors to fully differentiated, highly polarized neurons is extremely complex. Among the various phenomena
driving this process, those associated with membrane trafficking are of fundamental importance. The group is
interested in studying membrane trafficking in the very early steps of neuronal development. In this respect, they
have identified a process of developmentally-regulated, high-capacity endocytosis as one of the earliest marker of
neuronal polarity, and will investigate its function in neuronal commitment and axon pathfinding. This investigation
will be extended to the phenotypical characterization of the early steps of neuronal commitment of neural
stem/progenitor cells and iPS (induced-pluripotent stem cells). In the same cells, the late steps of neuronal
development will also be studied, i.e. the formation of synapses and their functional refinement, which are essential
phenomena for the proper integration of cells in neuronal networks.
The work in Grohovaz’s research unit is focused on the general comprehension of the physiopathological
mechanisms underlying neuroprotection or leading to neurodegeneration. The group mostly uses brain primary
cultures and applys molecular biology and in vitro cellular approaches, including live cell imaging (calcium imaging
and total internal reflection microscopy). Electrophysiology and animal behavioral studies are conducted in
collaboration with other research groups.
A main project deals with the generation of oxidative stress and the ensuing protection mechanisms in both neurons
and astrocytes. In particular, the mechanisms of iron entry and its influence on the production of reactive oxygen
species are investigated. On the other hand, in light of the importance of neuroinflammation, interest is also directed
to the role of glia in the neurodegenerative processes. More specifically, the molecular mechanisms involved in the
process of glia activation and the influence the activated phenotype has on neuronal survival are investigated.
Along with the study of these general physiopathological mechanisms, the group addresses specific issues within the
following disease-oriented projects:
(i) Alzheimer’s disease pathogenesis: the regulation of BACE1/BACE2 expression; the implication of beta-secretase
activity on amyloid-beta deposition; the cellular effects of the various amyloid-beta forms on different cell types from
brain.
(ii) Derangement in intracellular iron homeostasis and hereditary ferritinopathy: analysis of the mechanisms of iron
entry; cytotoxic effects of iron overload; cellular effects of ferritin light chain mutations.
(iii) Pathogenesis of neuronopathic (type A) Niemann-Pick disease: role of sphingosylphosphocholine on changes in
astrocytic phenotype; impairment of neuronal function.
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Synaptic disfunction and neuron-glia interplay
Research unit in Molecular Neuroscience
I.I.T. (Istituto Italiano di Tecnologia)
5 years (2008-2012); 600'000 EUR
Neurotransmitter release from nerve endings is the main form of information
transfer from one neuron to another or to an effector cell. Release occurs by
exocytosis from storage organelles, the synaptic vesicles, and is triggered by the
action potential-induced depolarization of the nerve terminal. The project pivots
on the idea that synapses perform an electro-chemical computation of both local
and environmental signals they receive. In this respect, a key role is played by
astrocytes, the close partners of neurons that, upon stimulation, release a
number of neurotransmitters and signaling molecules. The development of these
processes is investigated by developing and exploiting innovative molecular
imaging approaches.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Maurizio Popoli
Tel: +390250318361
E-mail: [email protected]
Center of Neuropharmacology, Department of Pharmacological Sciences,
CEND, University of Milan
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
1A. Presynaptic molecular machinery regulating glutamate release as
pathophysiological mechanism in amyotrophic lateral sclerosis
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Daniela Tardito
Tel: +390250318382
E-mail: [email protected]
Center of Neuropharmacology, Department of Pharmacological Sciences, CEND,
University of Milan
Proposta di ricerca
Area di interesse identificata
1A. Toward exploiting the screening potential of miRNome for the
identification of genes and pathways involved in neurodegenerative disorders
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
The regulation of neuroplasticity in the response to therapeutic drugs for mood
disorders. The role of micro RNAs
Cariplo Foundation
Durata progetto
March 2010-february 2013
Abstract del progetto
Aim of this project is to investigate, by means of an integrated approach that
merges preclinical and clinical studies with up-todate bioinformatic,
pharmacological, genomic and genetic techniques, the role of the posttranscriptional regulators miRNAs in Mood Disorders aetiology and
antidepressant response, with regard to neuroplasticity.
Overall, the main objectives of the present project will be:
1. to clarify the role of miRNAs in the molecular effects of different
antidepressants on the posttranscriptional modulation of genes coding for
modulators of neuroplasticity with an integrate approach based on preclinical
and clinical studies.
2. to discover new molecular targets for the development of innovative
therapeutic strategies for the treatment of drug resistant depression
3. to identify molecular markers associated to the antidepressant response in
human peripheral tissues for the optimization of drug treatment in mood
disorders. The integration of data obtained in clinical and preclinical studies will
permit the validation of blood leucocytes as cellular models for the study of
mental disorders
4. to identify new susceptibility genes and pharmacogenetic markers for
optimization of treatment in mood disorders.
Nome
Maria Pennuto, PhD
Contatti
Phone: +39 010 71781793, Fax: +39 010 720321
email: [email protected]
Italian Institute of Technology, Department of Neuroscience and Brain
Technology, Genoa 16163, Italy
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Mechanisms of Neurodegeneration
Identification of signaling pathways in neurodegenerative propeinopathies
Brain folding diseases, including Alzheimer’s disease, amyotrphic lateral sclerosis,
and polyglutamine (polyQ) diseases, are neurodegenerative disorders
characterized by the accumulation of toxic aggregation-prone proteins (Bertram
and Tanzi, 2005; Ross and Poirier, 2004; Taylor et al., 2002). PolyQ diseases are a
family of nine neurodegenerative disorders, including spinal and bulbar muscular
atrophy (SBMA). These disorders are caused by expansion of glutamine tracts in
nine genes, such as androgen receptor (AR). The mechanism through which the
mutation causes neurodegeneration is not known. Despite the ubiquitous
distribution of the disease proteins, neurons are especially vulnerable to
expanded polyQ. Moreover, specific populations of neurons are affected in each
polyQ disease. This suggests that the polyQ tract cannot be the sole determinant
of disease pathogenesis. Among polyQ diseases, SBMA represents an example
where the disease protein strikingly dictates disease features. SBMA is
characterized by the loss of lower motor neurons and skeletal muscle atrophy. In
SBMA, only males are fully affected. Gender-specificity is the result of the
conversion of polyQ AR into a toxic species induced by binding to its natural
ligand testosterone. AR is a hormone-activated transcription factor. The
mechanism through which ligand binding converts polyQ AR into a neurotoxic
molecule is unknown. Our long-term goal is to make a meaningful contribution
to our understanding of the molecular mechanisms of neurodegeneration in
brain folding diseases, so that treatment may be developed. Our objective is to
characterize the impact of protein context, cell context, and native protein
function on polyQ toxicity. We will test the central hypothesis that the toxicity of
polyQ AR is influenced at the post-translational level by phosphorylation of
specific residues in the disease protein, by tissue-specificity of disease protein
expression, and by aberrant protein function, e.g. interaction with transcription
co-factors and DNA. Greater insights into the molecular mechanisms underlying
the pathogenesis of polyQ diseases is important to developing novel and
effective therapeutic strategies for these diseases (reviewed by Pennuto and
Fischbeck, 2009).
Finanziamenti ricevuti
Titolo progetto
TARGETING ANDROGEN RECEPTOR FOR DEVELOPMENT OF SBMA THERAPEUTICS
Ente finanziatore
Durata progetto
Abstract del progetto
Muscular Dystrophy Association (USA)
01/07/2008 to 30/06/2011
The objectives of the MDA application are to identify post−translational
modifications, such as phosphorylation, ubiquitylation, and SUMOylation of AR
that reduce the toxicity of mutant protein, and to use this information to develop
effective therapeutic interventions. We propose to accomplish these objectives
by pursuing the following three specific Aims:
To determine the impact of protein kinase A phosphorylation, ubiquitylation, and
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
SUMOylation on the toxicity of mutant AR.
To evaluate the therapeutic efficacy of IGF−1 in vivo.
To select an effective system to deliver IGF-1 to SBMA mice.
POLYGLUTAMINE DISEASES: IMPACT OF PROTEIN AND CELL CONTEXT ON
NEUROTOXICITY
FP7−PEOPLE−2009−RG: Marie Curie International Reintegration Grant
01/04/2010 to 31/03/2014
The objectives of this proposal are to characterize the impact of three
components to polyglutamine toxicity: 1) protein context; 2) aberrant
protein−protein interactions; and 3) cell context. We will test
the central hypothesis that the toxicity of polyglutamine AR is influenced at the
post−translational level by phosphorylation of specific residues in the disease
protein, by aberrant interaction with the Akt effector FOXO, and by
tissue−specificity of disease protein expression.
INTERPLAY BETWEEN ANDROGENIC/ANABOLIC STEROID AND IGF-1 SIGNALING
IN AMYOTROPHIC LATERAL SCLEROSIS
Thierry Latran Foundation (France)
01/01/2011 to 31/12/2012
The objectives of this study are to i) Determine the effect of androgens and the
related anabolic steroid nandrolone in ALS pathogenesis; and ii) Characterize the
molecular cross−talk between androgens and
muscle−specific mIGF−1 in the maintenance of muscle phenotype.
INSULIN−LIKE GROWTH FACTOR 1/AKT AND ANDROGEN SIGNALING CROSSTALK
IN THE PATHOGENESIS OF SPINAL AND BULBAR MUSCULAR ATROPHY
Telethon-Italy
01/01/2011 to 31/12/2013
The objective of this application is to elucidate how phosphorylation of mutant
AR by Akt is regulated. The central hypothesis of this application is that
phosphorylation of mutant AR by Akt is altered by polyglutamine expansion,
ligand binding and age and that this modification is coordinated with other
post−translational modifications, including methylation and palmitoylation.
Nome
Tanganelli Sergio
Contatti
[email protected]
Istituto/Dipartimento
Dept. of Clinical and Experimental Medicine
Pharmacology Section, University of Ferrara, Italy
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Mechanisms of Neurodegeneration : A. Macromolecular interactions and
Neurodegeneration
Ente finanziatore
Neurotensin and neurotensin antagonist in an animal model of Parkinson’s
disease: therapeutic perspectives and role of NMDA/neurotensin interaction.
Sanofi-Aventis
Durata progetto
2007-2010
Abstract del progetto
The tridecapeptide neurotensin (NT) significantly enhances glutamatergic
signaling in different brain areas and amplifies the NMDA-receptor signaling,
probably by activating the high-affinity neurotensin receptor (NTS1), as shown by
the NTS1receptor antagonist SR48692-induced counteraction of this effect.
These findings suggest a reinforcing action of NT on several functions exerted by
glutamate in the central nervous system, in particular on the glutamatemediated excitotoxicity. NT could be implicated in the pathogenesis of chronic
and acute neurodegenerative disorders, such as Parkinson’s disease. To further
investigate this hypothesis, the aims of the project, which will combine
biochemical, functional, morphological and behavioral experiments, will be
evaluate: 1) the putative neuroprotective effects of a pretreatment with SR48692
(systemically administered) in an animal model of Parkinson’s disease [unilateral
6-hydroxydopamine (6-OHDA) lesion]; 2) the functional and physio-pathological
relevance of NMDA/NT receptor interactions in the striatum of naïve and 6OHDA -lesioned rats.
1. MECHANISMS OF NEURODEGENERATION
B. Neurotoxic stimuli
Nome
Domenico Garozzo
Contatti
[email protected]
0957338259 3473576202
CNR ICTP Catania
Via Gaifami 18 95125 Catania
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Glycomics
Finanziamenti ricevuti
Titolo progetto
CSF Glycomics in AD and other neurodegenerative diseases
Ente finanziatore
Durata progetto
Abstract del progetto
MIUR (Italy)
3 years
Protein glycosylation is the most common among post-translational
modifications (PTMs), which are key to the regulation of functional
activities of proteins. Quantitative and qualitative information about PTM
stages of proteins is crucial in the discovery of biomarkers of disease.
Macroheterogeneity of protein glycosylation depends on the extent of
glycosylation site occupancy while microheterogeneity refers to
differences in the sugar chain structures.
Differences of protein glycosylation may be indicative of species
synthesized in the central nervous system with respect to serum liverderived glycoforms, [as already demonstrated for protein Reelin, which is
implicated in Alzheimer disease and in other neurodegenerative disorders
(Botella-Lopez et al., 2006)]. Recent insights into neurodegenerative
disorders caused by abnormal glycosylation of protein Seipin
(Seipinopathy) show that modifications of Seipin glycosylation result in
improper folding leading to accumulation of the abnormal protein in the
endoplasmic reticulum, formation of ubiquitinated inclusion, and
activation of UPR (Ito and Suzuki, 2009).
Preliminary investigations based on lectin-blotting of CSF glycoproteins
revealed quantitative modifications in Alzheimer disease and altered
glycosylation of CSF transferrin (Taniguchi et al. 2008). We apply mass
spectrometry based glycomics of CSF as potential tool for to detect
possible macro-and microheterogeneity of CSF glycoproteins associated
to Alzheimer disease. The availability of CSF depository from patients
with AD and related disorders and established interactions with clinical
platforms will make possible to translate the results to clinical
proteomics.
The objective is to find out specific protein posttranslational
modifications related to glycosylation which in turn might be related to
the analysed cellular phenomena associated to neurodegeneration.
Nome
Maurizio Taglialatela, MD PhD – Professor of Pharmacology
Claudio Russo, PhD – Associate Professor of Pharmacoloy
Alfonso Di Costanzo MD – Associate Professor of Neurology
Giovanni Scapagnini MD PhD – Associate Professor of Clinical
Biochemistry
Contatti
Email: [email protected] Tel. +39-0874-404894/4851/4891
Istituto/Dipartimento
Dept. of Health Science, University of Molise, Via De Sanctis, 8610
Campobasso - ITALY
Proposta di ricerca
Area di interesse
identificata
Intracellular signalling during neurodegenerative events triggered upon APP
overexpression. The overexpression and the aberrant processing of the amyloid
precursor protein (APP) is a central event in the pathogenesis of Alzheimer’s Disease
(AD). Recent data suggest that the neuronal death that occurs in different
neurodegenerative conditions may arise from an aberrant signal related to cell cycle
re-activation in postmitotic neurons. We are currently investigating the role of APP
and of Aβ peptides as signalling molecules involved in MAPK and cell cycle
activation. We propose to study a specific molecular signalling involving APP and its
proteolytic processing by BACE1 and Presenilins, whose failure could explain at the
same time the formation of Aβ, the induction of cell cycle and neuronal dysfunction
observed in the development of AD.
• Venezia V. et al., Amyloid precursor protein and presenilin involvement in cell
signaling. Neurodegenerative Diseases. 2007, 4 (2-3): 101-111.
• Nizzari M. et al., Amyloid precursor protein and Presenilin1 interact with the
adaptor GRB2 and modulate ERK 1,2 signaling. The Journal of Biological Chemistry.
2007, 282 (18): 13833-13844.
Involvement of vascular factors as modulators of APP processing and signaling. The
complex interaction between vascular components and tissue homeostasis is
particularly relevant in the brain for the survival of vulnerable and irreplaceable
postmitotic neurons. It is debated whether the presence of vascular deposits of Aβ
peptides might represent a cause of oxidative stress and a predisposing condition
toward the development of dementia, or rather a protective endothelial mechanism.
In this context we are investigating the role of vascular components such as
thrombin and apolipoproteinE as modulators of endothelial processing of APP, in Aβ
formation and in Aβ-induced toxicity/protection.
• Russo C. et al., Opposite roles of apolipoprotein E in normal brains and in
Alzheimer’s disease. Proc. Natl. Acad. Sci, 1998, 95 15598-15602.
• Russo C. et al. Presenilin-1 mutations in Alzheimer's disease. Nature. 2000; 405
(6786):531-532.
• Russo C. et al., The amyloid precursor protein and its network of interacting
proteins: physiological and pathological implications. Brain Res Brain Res Rev. 2005
48 (2): 257-264.
Role of specific classes of ion channels in AD-related neurodegeneration.
Intracellular potassium concentrations play a key role in cell survival. A decrease in
cytoplasmic [K+]i, mainly caused by an increased activity of plasma membrane
voltage-gated potassium channels, triggers cell death. Changes in K+channel activity
play a major pathogenetic role in many neurodegenerative disorders, including
Alzheimer’s
disease (AD). In fact, β-amyloid fragments (Aβ) alter the properties of K+ currents in
mammalian neurons. We have described a selective modulation of a neuronal K+
channel (Kv3.4) upon Aβ exposure and we have shown that the inhibition of this
channel prevents cell death triggered by the neurotoxic stimulus. We are currently
investigating the potential neuroprotective role of Kv3.4 blockers in several models
of AD-related neurodegeneration..
• Pannaccione A., et al. Nuclear factor-kappaB activation by reactive oxygen
species mediates voltage-gated K+ current enhancement by neurotoxic betaamyloid peptides in nerve growth factor-differentiated PC-12 cells and
hippocampal neurones. J Neurochem. 2005 Aug;94(3):572-86.
• Pannaccione A, et al. Up-regulation and increased activity of KV3.4 channels and
their accessory subunit MinK-related peptide 2 induced by amyloid peptide are
involved in apoptotic neuronal death. Mol Pharmacol. 2007 Sep;72(3):665-73.
Oxidative stress as a pathogenetic mechanisms and therapeutic target for
Alzheimer Disease. Reduction of cellular expression and activity of antioxidant
proteins leading to oxidative stress are fundamental causes for brain aging and
neurodegenerative diseases. Deregulation of the antioxidant protein Heme
oxygenase-1 (HO-1) has been associated with the pathogenesis of Alzheimer's
disease, multiple sclerosis and brain aging. Moreover, a number of experimental and
epidemiological studies have recently supported the beneficial effects of some
commonly used natural products such as curcumin, possibly by increasing the
activity of the HO-1 protein. Curcumin has been reported to decrease oxidative
damage and amyloid deposition in a transgenic mouse model of Alzheimer's disease,
and to reverse Aβ-induced cognitive deficits and neuropathology in rats. We are
currently investigating the potential neuroprotective role and the underlying
mechanisms, of HO-1-inducing natural compounds in various in vitro and in vivo
models of AD.
• Scapagnini G. et al., Caffeic acid phenethyl ester and curcumin: a novel class of
hemeoxygenase-1 inducers. Mol Pharmacol. 2002, 61(3):554-61.
• Scapagnini G. et al., Ethyl ferulate, a lipophilic polyphenol, induces HO-1 and
protects rat neurons against oxidative stress. Antioxid Redox Signal. 2004, 6(5):8118.
• Scapagnini G. et al., Curcumin activates defensive genes and protects neurons
against oxidative stress. Antioxid Redox Signal. 2006, 8(3-4):395-403.
Physical exercise and cognitive decline. Epidemiological studies have shown that
physical exercise can delay the occurrence and the progression of dementia, but also
to improve physical, cognitive and psychological performances, with a positive
impact on the quality of life. The mechanisms involved and the identification of
potential biomarkers predictive of a positive response to physical activity are areas
of intense investigation. In our group, we are currently investigating the potential
preventive role of physical exercise programs on cognitive decline in subjects at risk
to develop dementia. Our focus will be on subjects presenting with a subjective
memory disturbance or affected by mild cognitive impairment (MCI).
• Tedeschi G., et al. Brain atrophy and lesion load in a large population of patients
with multiple sclerosis. Neurology. 2005 Jul 26;65(2):280-5.
• Tedeschi G., et al. Correlation between fatigue and brain atrophy and lesion load
in multiple sclerosis patients independent of disability. J Neurol Sci. 2007 Dec
15;263(1-2):15-9.
Finanziamenti ricevuti
Titolo
Regulation of K+ channels by APP-interacting proteins: functional characterization and
progetto
implications for AD-related neurodegeneration
Ente
finanziatore
Durata
progetto
Abstract del
progetto
Titolo
progetto
Regione Campania (2000-2003)
3 years
Recent experimental evidence suggest that K+ channels play a major pathogenetic role in
Alzheimer disease. In the CNS, phosphorylation of voltage-gated K+ channels by tyrosine
kinases (PTK) is a fundamental process regulating their functional activity; its inhibition
exerts a neuroprotective action in several experimental models of neuronal damage. On
the other hand, the expression of several subclasses of K+ channels can reduce cellular
tyrosine kinase activity, thus suggesting the existence of a reciprocal interaction between
PTKs/PTPases and K+ channels. Given that the cytosolic domain of β-APP influences,
through its adaptor proteins Dab (disabled), X11 and Fe65, the tyrosine kinase activity of
Abl, in the present proposal we will:
A. Evaluate the potential modulation of several classes of K+ channels (Kv1.2, Kv1.3, Kv1.5,
Kv2.1, ERG, KCNQ2+3) by adaptor proteins able to bind to β-APP (X11, Fe65, Dab).
B. Verify the participation of tyrosine kinase activity (and in particular of Abl), in this
modulation.
C. Investigate the potential involvement of these channels in the cellular damage induced
by the hyperexpression of β-APP adaptor proteins.
D. Study the participation of K+ channels and their regulation by oxidative stress in the
neuronal death triggered by various cellular models of AD-related neurodegeneration.
European Community contract N° LSHM-CT-2003-503330/Apopis : Abnormal proteins in
the pathogenesis of neurodegenerative disorders
Study on tyrosine phosphorylation of APP and its interaction with intracellular adaptors:
role in cell signaling and in the generation of amyloidogenic fragments.
Ente
finanziatore
Durata
progetto
Abstract del
progetto
European Community
3 years (2004-2007)
Within the WP 1 Generation and Turnover, our proposal is aimed to the definition of the
role that posttrasductional modifications of APP, in particular its phosphorylation, may
exert on its pathophysiologycal function. We will investigate both the role of
phosphorylated APP/CTFs in cell signaling, and/or in the generation of amyloidogenic
fragments and plaques through the interaction with intracellular adaptors, as well as the
effect that site-specific tyr-phosphorylation would have on the direct amyloidogenic
pathway of APP. These studies, which are focused in a completely new aspect of APP
activity, would lead to the identification of early markers of the neurodegenerative process
which occurs in AD, and to the definition of a potentially innovative therapeutical approach
based on the findings obtained. Our proposal is aimed at:
Aim 1) To study directly in human brain the tyr-phosphorylation of APP and its CTFs and
their possible coupling with intracellular adaptors such as ShcA, and to correlate these data
with the presence of neuropathologycal hallmarks of the disease. The comparative analysis
in non-AD control, AD and/or Down’s syndrome subjects at different ages (from fetal to
adult life) will allow a deeper characterization of the molecular determinants during the
progression of the disease.
Aim 2) To study in transgenic mice carrying APP and PS1 mutations the tyrphosphorylation of APP and CTFs, their interaction with ShcA-Grb2 adaptors and the
influence that such interaction could have on plaque formation, astroglial response,
neuronal death. The use of a Tg model will allow at a deeper comprehension of the
possible correlation between APP phosphorylation, its cleavage, signaling activity and
progressive presence of the typical hallmarks of the disease. Moreover, in this model is
possible also to determine the effect that APP and PS1 mutations, which are linked with a
human familial phenotype, would cause in the above mentioned parameters, and, in a
second step, would allow the application of targeted therapeutical pharmacological
approaches following also the indications raised from the in vitro studies (aim 3).
Aim 3) To study in neuronal and/or glial cell cultures the molecular mechanisms which
regulate APP phosphorylation, influence on cleavage by secretases, coupling with
intracellular adaptors, and signaling activity. Using APP mutants, ShcA mutants, and APP
KO cells we will define the influence that phosphorylation and interaction with ShcA would
cause in signaling activity, cell proliferation and death, and in the generation of
amyloidogenic fragments. Moreover, the study of the effect that proliferative and
apoptotic stimuli may cause on those parameters and/or with the study of the kinases
involved in the activation of APP cleavage and signaling, this would also lead to the
identification of pharmacological target for a therapeutical intervention.
Nome
Contatti
Istituto/Dipartimento
MARIO MARCHI
[email protected]
Department of Experimental Medicine (Di.Me.S.)
Section of Pharmacology and Toxicology
Viale Cembrano, 4 16147 Genova ITALY +39 010 3532657 e.mail:
Proposta di ricerca
Effect of Age and neurodegenerative disorders on central neurotransmission: focus on neurotransmitter
release from neurons and glia
The effects of age and/or neurodegenerative disorders may differentially influence diverse cell types in
the brain and the vulnerability of neurons may depend also on the transmitter system or the location in
neural circuits. Therefore, it is of utmost importance to identify age-induced neurochemical changes in the
brain in order to develop pharmacological strategies that can prevent, delay or counteract the cognitive
decline.
Cells may be differentially sensitive according to the function of signaling pathways that mediate cell
specific processes involved in cognition (e.g. synaptic plasticity). Systemic influences of inflammation can
also alter the local milieu to influence different cell populations. Our current researches are focused to
assess the current status of synaptic function/dysfunction in aging and age-related neurodegenerative
diseases in several regions of the brain in mammals, particularly in rodents .It is now well accepted that
astrocytes integrate and process synaptic information and control synaptic transmission and plasticity
being active partners in synaptic function, astrocytes are cellular elements involved in the processing,
transfer and storage of information by the nervous system. Therefore our studies are aimed at
investigating “ex-vivo” changes of neuronal and glial functions due to “in-vivo” aging or in the animal
models of neurodegenerative disorders (i.eAlzheimer;) The pivotal concept of this approach assumes that
“in-vivo” aging or the neurodegenerative disorders are able to modify the plasticity of chemical synapses
which in turn could be retained, and then analyzed, in the “ex-vivo” tissue preparations. This “ex-vivo”
memory has been already described to occur and studied in our laboratory to unmask neurotransmission
modifications caused by “in-vivo” exposure to different stimuli.( i.e. chronic treatment with drugs,
enriched environment, etc.). Our research is therefore focused to investigate whether and to what extent
aging and age-related neurodegenerative diseases (i.e. Alzheimer) could modify some fundamental
parameters, such as neurotransmitter release and uptake, receptor expression and functioning in plasma
membranes, moreover we will investigate the effect of beta amyloid on these parameters by utilizing two
“ex-vivo” functional preparations representative of neurons and astrocytes , i.e. the isolated nerve
terminals (synaptosomes) and the glial subcellular particles (gliosomes).
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Plasticità dei recettori presinaptici nicotinici e glutammatergici che
modulano la liberazione di neurotrasmettitori: studio delle variazioni
delle loro risposte funzionali dopo trattamenti cronici o in seguito ad
interazioni con altri sistemi recettoriali o altre condizioni sperimentali
Role: Principal Investigator
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
MIUR-prin
24 MESI
Studies on the effect of the chronic treatment with nicotine on some
neurochemical presynaptic mechanisms and on the function of
presynaptic receptors in the CNS.
Role: Principal Investigator
Ente finanziatore
Durata progetto
Titolo progetto
COMPAGNIA DI SAN PAOLO
36 MESI
Les plant aromatique, entre environment et activitè productive
Role . Principal Investigator
Ente finanziatore
Durata progetto
ALCOTRA “AROMA”
24 MESI
Nome
Contatti
Dr. Francesca Ruberti
[email protected]; INMM- CNR Istituto di Neurobiologia e Medicina
Molecolare Via del Fosso di Fiorano, 64 00143 ROMA ITALY Phone: + 39
06 501 703 236 Fax: + 39 06 501 703 313
Istituto/Dipartimento
Institute of Neurobiology and Molecular Medicine CNR
Proposta di ricerca
NEUROBIOLOGY OF ALZHEIMER’S DISEASE
1) Basic research : developing competitive in vitro and in vivo models to study Alzheimer’s Disease
a) Role of microRNAs in Alzheimer’s Disease onset and progression (C. Barbato, C Cogoni, F. Ruberti)
Primary neuronal cell culture and Alzheimer’s Disease (AD) mouse models, including AD11 (and transgenic
mice overexpressing pathogenic variants of human APP gene, will be instrumental to investigate the
function of selected miRNAs (by their overexpression and/or inhibition) on disease pathogenesis. Studies
will focus on recently identified microRNAs modulating, in neuronal hippocampus cultures (Vilardo et al.
2010), APP production and on miRNAs dysregulated in human AD brain (Barbato et al 2009).
Multidisciplinary approaches will be used to investigate the effect of microRNAs on molecular mechanisms
underlying amyloidogenesis, neuronal degeneration, memory loss, synaptic plasticity impairment.
Vilardo E, Barbato C, Ciotti MT, Cogoni C, Ruberti F. MicroRNAs regulate Alzheimer’s Amyloid Precursor
Protein expression in hippocampal neurons. (2010) submitted.
Barbato C, Ruberti F, Cogoni C. Searching for MIND: microRNAs in neurodegenerative diseases. J Biomed
Biotechnol. 2009:871313.
Barbato C, Ruberti F, Pieri M, Vilardo E, Costanzo M, Ciotti MT, Zona C, Cogoni C. MicroRNA-92 modulates
K(+) Cl(-) co-transporter KCC2 expression in cerebellar granule neurons. J Neurochem. 2009 Dec 26.
b) Mechanisms underlying the effect of N-terminal 26-230 tau fragment on synaptic dysfunction and
neurodegeneration (N. Canu, in collaboration with V. Cestari, F. Tirone, E Mattei). The aim is is to decode
the signal transduction pathway that links N-terminal 26-230 tau fragment (N-tau) expression to NMDAR
activity (Canu et al 1998; Amadoro et al 2006) and their upstream and downstream effectors inducing
synaptotoxicity and plasticity failure. Electrophysiological recording of NMDAR and AMPAR biochemical
characterization of NMDAR complex, morphological, qualitative and quantitative analysis of synapse
number, dendritic spines and behavioral analysis of conditional N-tau transgenic mice, will be performed
to give an accurate and global view of the molecular events involved in N-tau modulation of neuronal
plasticity.
Canu N, Dus L, Barbato C, Ciotti MT, Brancolini C, Rinaldi AM, Novak M, Cattaneo A, Bradbury A, Calissano
P. Tau cleavage and dephosphorylation in cerebellar granule neurons undergoing apoptosis. J Neurosci.
1998 Sep 15;18(18):7061-74.
Amadoro G, Serafino AL, Barbato C, Ciotti MT, Sacco A, Calissano P, Canu N. (2004) Role of N-terminal tau
domain integrity on the survival of cerebellar granule neurons. Cell Death Differ. 2004 Feb;11(2):217-30.
Amadoro G, Ciotti MT, Costanzi M, Cestari V, Calissano P, Canu N. NMDA receptor mediates tau-induced
neurotoxicity by calpain and ERK/MAPK activation.Proc Natl Acad Sci U S A. 2006 103(8):2892-7
c) Nerve Growth Factor deprivation pathways leading to Alzheimer’s Disease phenotype (C. Matrone, P.
Calissano). An NGF-dependent hippocampal neuronal model in which the interruption in NGF supply
quickly triggers amyloidogenesis and induces tau protein dysfunction and neuronal degeneration,
mimicking essential features of in vivo AD pathology, has been recently charachterised (Matrone et al.
2008; Matrone et al 2009; Amadoro et al 2009). The main goal is to to test whether the proteins whose
expression or phosphoylation is affected by NGF deprivation may represent early molecular signature and
potential targets of AD. To achieve these results, studies are in progress, in collaboration with prof. Moses
Chao at NYU, to characterize the phosphotyrosine-associated signalling events due to NGF deficit in
primary cultures from wild type and APPk/o mice by a systematic MS/SILAC.
Matrone C, Ciotti MT, Mercanti D, Marolda R, Calissano P. NGF and BDNF signaling control amyloidogenic
route and Abeta production in hippocampal neurons.Proc Natl Acad Sci U S A. 2008 105:13139-44.
Matrone C, Marolda R, Ciafrè S, Ciotti MT, Mercanti D, Calissano P. Tyrosine kinase nerve growth factor
receptor switches from prosurvival to proapoptotic activity via Abeta-mediated phosphorylation. Proc Natl
Acad Sci U S A. 2009 106: 11358-63.
Amadoro G, Corsetti V, Ciotti MT, Florenzano F, Capsoni S, Amato G, Calissano P. Endogenous Abeta causes
cell death via early tau hyperphosphorylation. Neurobiol Aging. 2009 Jul 21.
d) Role of NH2-26-44 tau peptide in Alzheimer’s Disease (G. Amadoro P Calissano). A synthesized NH2-2644 peptide of tau, impairs the mitochondrial oxidative phosphorylation and reduces the intracellular ATP
bioavailability, probably acting on adenine nucleotide translocator (ANT)- mediated ADP/ATP exchange
(Atlante et al., 2008). Studies in AD human tissues are focused to evaluate if tau peptide i) is correlated
with the extent of neurofibrillary degeneration and amyloid neuropathology, especially Abeta pre-fibrillar
species; (ii) is linked to the synaptic changes and to the mitochondrial functional alterations. The physical
interaction between ANT and NH2-derived tau fragment, in pathologically relevant conditions, will be
investigated.
Atlante A, Amadoro G, Bobba A, de Bari L, Corsetti V, Pappalardo G, Marra E, Calissano P, Passarella S. A
peptide containing residues 26-44 of tau protein impairs mitochondrial oxidative phosphorylation acting at
the level of the adenine nucleotide translocator. Biochim Biophys Acta. 2008 1777, 1289-300
e) Mitochondrial damage and genetic risk/susceptibility factors in Alzheimer’s Disease (M. Rinaldi, S.
Iurescia, D. Fioretti). The aim of the research project is i) to identify and characterize early biomarkers and
mechanisms of mitochondrial damage promoted by ROS/RNS; ii) to investigate the protective role of
natural forms and synthetic analogues of vitamin E that can be tested in intervention studies in AD
patients. Studies are in progress to evaluate post-transductional modifications (PTMs) in mitochondrial
proteins, caused by ROS/RNS in in vitro model of senescence and human neuroblastoma as well as in
peripheral cells from subjects with Mild Cognitive Impairment (MCI) and AD. Strong genetic
risk/susceptibility factors for developing Alzheimer’s disease, such as ApoE4 that impacts on Abeta
production, Abeta clearance, Abeta fibrillation and tangle formation as well as on mitochondrial functions
leading to neuronal toxicity and synaptic damage, are also investigated.
Iurescia S, Fioretti D, Mangialasche F, Rinaldi M.The Pathological Cross Talk Between Apolipoprotein E and
Amyloid-beta Peptide in Alzheimer's Disease:Emerging Gene-Based Therapeutic Approaches. J Alzheimers
Dis. 2010 In press
2) Basic research : New treatment strategies based on Neurotrophins
f) Ocular NGF administration as a novel non invasive approach to protect brain-NGF target neurons that
degenerate in Alzheimer’s disease (L. Aloe, P. Tirassa). Nerve growth factor and (NGF) is a soluble protein
that plays a protective role on brain neurons that degenerate in age-related brain disorders, including
Alzheimer’s disease (AD). We have shown that conjunctively applied NGF can protect injured brain
neurons and damaged retinal ganglion neurons suggesting that topical eye NGF application might be a
novel alternative for delivering NGF into the brain and for protecting brain neurons and reducing memory
deficits occurring during early events of AD and glaucoma. This hypothesis is currently actively under
investigation in collaboration with other research groups.
Lambiase A, Pagani L, Di Fausto V, Sposato V, Coassin M, Bonini S, Aloe L. Nerve growth factor eye drop
administrated on the ocular surface of rodents affects the nucleus basalis and septum: biochemical and
structural evidence. Brain Res. 2007;1127:45-51.
Di Fausto V, Fiore M, Tirassa P, Lambiase A, Aloe L. Eye drop NGF administration promotes the recovery of
chemically injured cholinergic neurons of adult mouse forebrain. Eye drop NGF administration promotes
the recovery of chemically injured cholinergic neurons of adult mouse forebrain. Eur J Neurosci.
2007;26:2473-80.
Lambiase A, Aloe L, Centofanti M, Parisi V, Mantelli F, Colafrancesco V,
Manni GL, Bucci MG, Bonini S, Levi-Montalcini R. Experimental and clinical evidence of neuroprotection by
nerve growth factor eye drops: Implications for glaucoma. Proc Natl Acad Sci U S A, 2009; 109: 1346913474,.
g) Electro-acupuncture and polyphenols as novel approaches to reduce neurodegeneration ( L Manni, M
Fiore). Preliminary data indicate that diabetes induced in adult rats by injection with streptozotocin causes
a decrease of brain NGF and a concomitant increase of tau phosphorylation, that were all counteracted by
low-frequency electro-acupuncture (EA). Using molecular and behavioural approaches the research
project will be aimed at: i) Investigate the possible link between diabetes, brain NGF presence/activity and
the development of AD-associated pathological features; ii) Study the effect of EA on the above mentioned
diabetes-associated brain dysfunction. Alcoholism is related to neurodegeneration and diabetes. The use
of polyphenols extracted by olive oil, olive leaves and olive seeds or by grapes (Fiore et al 2009) to prevent
or limit mouse brain neurodegeneration due to aging or ethanol consumption will be evaluated.
Manni L, Aloe L, Fiore M. Changes in cognition induced by social isolation in the mouse are restored by
electro-acupuncture. Physiol Behav. 2009;98(5):537-42
Fiore M, Laviola G, Aloe L, di Fausto V, Mancinelli R, Ceccanti M. Early exposure to ethanol but not red wine
at the same alcohol concentration induces behavioral and brain neurotrophin alterations in young and
adult mice. Neurotoxicology. 2009;30(1):59-71.
h) Human neural stem cells lentivirally transduced with human BDNF (C. Cenciarelli, P. Casalbore). A
strong body of evidences show that BDNF result critically involved in Alzheimer’s and Huntington’s
diseases. Recently, it has been shown that BDNF knockdown within NSC abolishes the cognitive benefits of
NSC delivery. The aim is to use propagating human neural stem cells (hNSC) lentivirally transduced with
human BDNF as cellular therapy for replacing degenerating neurons in disease, trauma and toxic insults.
Cenciarelli C, Budoni M, Mercanti D, Fernandez E, Pallini R, Aloe L, Cimino V, Maira G, Casalbore P. In vitro
analysis of mouse neural stem cells genetically modified to stably express human NGF by a novel multigenic
viral expression system. Neurol Res. 2006, 28:505-12.
Casalbore P, Barone I, Felsani A, D’Agnano I, Michetti F, Maira G and Cenciarelli C. Neural stem cells
modified to express BDNF antagonize trimethyltin-induced neurotoxicity through PI3K/Akt and MAP kinase
pathways . Accepted on J. Cell. Physiol. 2010.
OTHER NEURODEGENERATIVE DISEASES
Basic research, biobanking (DNA), sharing database, exchanging and pooling data
i) Biobank compund by over 2000 DNAs from HD and ataxic patients and their families. Molecular basis
of episodic ataxia (L. Veneziano). Research activity has been mostly devoted to CACNA1A gene, coding for
the Alpha1A subunit of P/Q type Calcium channel expressed in brain, from its physical map to the
refinement of the gene structure to the screening for mutations in Episodic Ataxia (EA) to the association
genotype/phenotype (Veneziano et al 2009; Mantuano et al 2010). This research allowed the construction
of a DNA biobank from about 100 patients affected by EAs. The project is focused on the use Comparative
Genomic Hybridization (CGH) arrays, which can be used for a screening of CACNA1A gene rearrangement
in a sample of 40 EA patients in which CACNA1A gene point mutations were excluded. The fulfilment of
the above aim will improve the rate of molecularly diagnosed EA2 patients to be treated with known
therapy or to be included in new therapeutic trials.
Veneziano L, Guida S, Mantuano E, Bernard P, Tarantino P, Boccone L, Hisama FM, Carrera P, Jodice C,
Frontali M. Newly characterised 5' and 3' regions of CACNA1A gene harbour mutations associated with
Familial Hemiplegic Migraine and Episodic Ataxia. J Neurol Sci. 2009, 276(1-2):31-7
Mantuano E, Romano S, Veneziano L, Gellera C, Castellotti B, Caimi S, Testa D, Estienne M, Zorzi G, Bugiani
M, Rajabally YA, Barcina MJ, Servidei S, Panico A, Frontali M, Mariotti C.Identification of novel and
recurrent CACNA1A gene mutations in fifteen patients with episodic ataxia type 2. J Neurol Sci. 2010 Feb 1.
l) Role of the Thyroid Transcription Factor 1 (TTF-1) in neurodegenerative diseases (D. Civitareale). The
Thyroid Transcription Factor 1 (TTF-1) expression persists in some post-mitotic basal forebrain neurons,
and mutations of TiTF-1 have been associated with the benign hereditary chorea. Our transcriptome
analysis confirms that TTF-1 is involved in neuron morphogenesis, differentiation and GnRH secretion. We
predict that TTF-1 could be a modifier gene involved in the pathogenesis of the neurodegenerative
diseases. Therefore, we propose to study TTF-1 down-regulation via RNAi and microarray analysis in
ST14A, select a mouse strain able to develop the neurodegenerative disease in absence of Titf-1, and
characterize the genotype/phenotype association of TiTF-1 in a cohort of patient with neurodegenerative
diseases.
Provenzano C, Veneziano L, Appleton R, Frontali M, Civitareale D. Functional characterization of a novel
mutation in TITF-1 in a patient with benign hereditary chorea. J Neurol Sci. 2008 264(1-2):56-62.
Provenzano C, Pascucci B, Lupari E and Civitareale D. Large scale analysis of transcription factor TTF1/NKX2.1 target genes in GnRH secreting cell line GT1-7. Molecular and Cellular Endocrinology. In press.
Area di interesse identificata
Basic research : developing competitive in vitro and in vivo models to
study Alzheimer’s Disease; new treatment strategies based on
Neurotrophins.
Biobanking
(DNA), sharing database, exchanging and pooling data
Finanziamenti ricevuti Role of microRNAs in Alzheimer’s Disease onset and progression (C. Barbato, C
Cogoni, F. Ruberti,).
Ongoing Research Support
-3886
SD/sd 2008.2404 Programma Neuroscienze “Compagnia di San Paolo” 9/01/09-9/01/11:
‘MIND: MicroRNA In Neurodegenerative Diseases’ (to C. Barbato)
-REGIONE LAZIO grant 2009-2011: ‘Studio delle basi molecolari della neurodegenerazione
nella malattia di Alzheimer’- (to C.Cogoni).
Completed Research Support
DG.RSTL.059.012 –CNR grant-01/2008-12/2009. “Study of APP regulation by microRNAs
and implication in Alzheimer’s Disease” (to F. Ruberti)
Finanziamenti ricevuti Mechanisms underlying the effect of N-terminal 26-230 tau fragment on synaptic
dysfunction and neurodegeration (N. Canu).
Ongoing Research Support
Dipartimento di Neuroscience, University of Tor Vergata
Completed Research Support
MIUR-PRIN 2006-2008: “Molecular and behavioural analysis of tau neurotoxic effect in
cellular and animal models”
-Ministero della Sanità- Ex Articolo 56 Neurodegenerativo 533F/B/1 2005-2009
“Identificazione di marcatori biologici precoci per la demenza di Alzheimer: genomica e
proteomica nella neurodegenerazione”
Finanziamenti ricevuti Nerve Growth Factor deprivation pathways leading to Alzheimer’s Disease
phenotype (C. Matrone, P. Calissano).
Ongoing Research Support
MIUR-FIRB Italy / 2009-2011
“Italian Human Proteome Net”
MIUR-PRIN 2009/2011
“Ruolo di Zn e Cu nella cascata amiloidogenica provocata da deprivazione di NGF”
Finanziamenti ricevuti Role of NH2-26-44 tau peptide in Alzheimer Disease (G. Amadoro P. Calissano)
Ongoing Research Support
MIUR-FIRB 24-09-08 al 24-09-11: “Italian Human Proteome Net”
MIUR 18-07-2007 to 18-07-2010: “Neurotrophic and neuroprotective drugs suitable for
therapeutic applications in neuroscience: AD,HD,SLA”
Finanziamenti ricevuti Mitochondrial damage and genetic risk/susceptibility factors in Alzheimer’s
Disease (M. Rinaldi, S. Iurescia, D. Fioretti).
Ongoing Research Support
MIURPRIN 2007-2009: “Oxidative and nitrosative damage of mitochondrial proteins in models
of cell senescence and in subjects with Alzheimer's disease: a pre-clinical study on natural
forms and new synthetic analogues of vitamin E.
Finanziamenti ricevuti, Ocular NGF administration as a novel non invasive approach to protect brainNGF target neurons that degenerate in Alzheimer’s disease (P Tirassa, L Aloe)
Ongoing Research Support
Progetto PRIM- 2007-2010: “Sviluppo del NGF come farmaco in collirio: Studi di
farmacocinetica e di efficacia terapeutica”. (to L. Aloe)
Progetto Strategico Q77, ISS- 2007-2010: “Indicazioni di indici neurobiologici di
vulnerabilità a rischio di sviluppo di sindromi depressive” (to L. Aloe).
Fondazione G.B. Bietti 2007-2009: “Potenziale ruolo terapeutico del NGF nel glaucoma:
Studi in un modello animale e nell’uomo” (to L. Aloe)
CNR funding (PORRSTL03) 2006-2008 (to P. Tirassa)
Finanziamenti ricevuti, Electro-acupuncture and polyphenols as novel approaches to reduce
neurodegeneration (L Manni, M Fiore)
Ongoing Research Support
Centro
Alcologico Regione Lazio (to M. Fiore)
Finanziamenti ricevuti, Human neural stem cells (hNSC) lentivirally transduced with human BDNF (C.
Cenciarelli, P. Casalbore).
Ongoing Research Support
Funds
from ATENA Onlus (2009-2010): “Transplantation of BDNF secreting-NSC in the
hippocampus of adults rats treated with the neurotoxicant thrimetyltin.”
Completed Research Support
Grants
from The Nando Peretti Foundation and ATENA Onlus (2006-2008)
“Repair of
lesions of Central Nervous System through cell replacement therapy by using of NSC
induced towards to neuronal phenotypes.”
Finanziamenti ricevuti, Biobank compund by over 2000 DNAs from HD and ataxic patients and their
families and Molecular basis of episodic ataxia (L. Veneziano).
Ongoing Research Support
National
Ataxia Foundation 2009: “Looking for gross rearrangements of CACNA1A gene in Episodic
Ataxia type 2 (EA2) patients by Comparative Genomic Hybridization arrays.
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
MARIAROSARIA MILOSO
[email protected] tel 0264488123
Dipartimento di Neuroscienze e Tecnologie Biomediche
EVALUATION OF MESENCHYMAL STEM CELLS (MSCs) EFFECT IN
ALZHEIMER’S DISEASE RAT MODELS: in vitro and in vivo STUDIES
Alzheimer’s disease (AD) is the most common cause of progressive
decline of cognitive function in aged humans. At present, there are no
effective therapies to treat this pathology. Recently it has been
hypothesized an innovative therapeutic approach based on the use of
stem cells. Different sources of stem cells are being explored for potential
use
in
repairing
neurodegenerative
disorders.
Among
them,
Mesenchymal Stem Cells (MSCs) represent a good cellular source for
transplantation therapy. MSCs can be easily isolated and expanded,
autologously used, present a low risk of tumorigenicity and are
ipoimmunogenic.
Aim of our project is to study the possible effect of human and rat MSCs
in an in vitro and in vivo model of Alzheimer Disease (AD).
In vitro model of AD is represented by rat cortical and hippocampal
neuron primary cultures exposed to 2,5 µM Aβ (1-42) fibrils. To evaluate
the capacity of MSCs to improve neuronal viability and AD
neuropathological changes, we will perform: a) direct and indirect cocultures of MSCs with Aβ-treated neurons; b) culture of Aβ-treated
neurons in the presence of conditioned medium from MSCs. Neuronal
survival will be evaluated by MTT assay, neuronal morphology and
functionality will be assessed by morphological and immunofluorescence
studies and electron microscopy.
In vivo model of AD is represented by a transgenic rat model that develop
spontaneously the AD pathology. The MSCs are prelabeled with Qdot
nanocristal and than inoculated into the rat tail vein. After the MSCs
injection cognitive functions will be assessed by the Morris water maze
test (Morris 1984). After the behavioral test, the rats will be sacrificed
and the brain will be obtained and further fixed. Sections of cortex and
hippocampus
will
be
cut
in
a
freezing
microtome
and
AD
neuropathological changes will be evaluate by morphological and
immunofluorescence studies and electron microscopy. MSC localization
and
differentiation will be evaluated by immunofluorescence
experiments.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Nome
Patrizia Hrelia, Full Professor in Toxicology, Faculty of Pharmacy
Contatti
Via Irnerio 48, 40126 Bologna
Tel.: +39 051 2091799
e-mail: [email protected]
Istituto/Dipartimento
Department of Pharmacology , Alma Mater Studiorum - University of
Bologna
Nome
Silvana Hrelia, Associate Professor in Biochemistry, Faculty of
Pharmacy
Contatti
Via Irnerio 48, 40126 Bologna
Tel.: +39 051 2091233
e-mail: [email protected]
Istituto/Dipartimento
Department of Biochemistry, Alma Mater Studiorum - University of
Bologna
Proposta di ricerca
Our project is aimed at the identification of cellular and molecular targets affected by
phytochemicals, to delineate pharmacological and nutritional strategies for the reduction or the
prevention of neurodegeneration and neuroinflammation associated to ageing and
neurodegenerative diseases. The project is innovative since it represents a joint action with the
attempt:
1) to adequately understand the cellular and molecular mechanisms commanding cell death in
neurodegeneration and behind neuroprotective effects of phytochemicals;
2) to identify phytochemicals having diverse pharmacological activities, such as neuroprotection and
antiinflammation;
3) to confirm if these phytochemicals have a specific pharmacological effectiveness in the
prevention/counteraction of one of the most important aging-related neurodegeneration and
neuroinflammation hallmarks, oxidative stress, as a primary prevention mechanism;
4) to understand if these phytochemicals can act as biomodulators in neuronal cells through
modulation of survival pathways, suggesting a potential therapeutic use in regenerating injured
neuronal cells;
5) to establish the therapeutic window that allows an effectively impact of phytochemicals on
specific targets;
In longer terms, our studies will indicate whether pharmacological and nutritional strategies with
phytochemicals should be effective and safe in counteracting cellular and molecular damage
associated to human neurodegeneration and neuroinflammation.
Area di interesse identificata
New treatment strategies (Neurodegeneration and
Neuroprotection)
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Drug Research and Development (CHEM-PROFARMA-NET).
Synthesis, biological and pharmacological characterization of
organic molecules, bio-oligomers and natural compounds endowed
with antidegenerative (neuro or cardiovascular) antiviral, and antiinfective activity.
MIUR FIRB – Piattaforme/Reti (total funds euro9.768.680,
euro1.274.097 assigned to P.Hrelia as node coordinator)
Durata progetto
2007-2011
Abstract del progetto
The vision of the project is that of a strong collaborative efforts
aimed at the identification of new cellular and molecular targets
affected by synthetic or natural compounds to delineate
pharmacological and nutritional strategies for the prevention of
human diseases such as neurodegeneration, infectious disease
cancer, immune and metabolic disorders. The activities proposed
will be completely performed in the Network Operative Units. This
project is aimed at identifying new drugs that may be useful in
preventing and/or alleviating the consequences of specific diseases,
but it will focus on new therapeutic approaches for such disorders.
Titolo progetto
New Bioactive Molecules from Natural Sources in Neuroprotection:
Opportunities In Neurodegenerative Diseases
Ente finanziatore
MIUR COFIN 2007 (total funds euro 63.750 assigned to G. Cantelli
Forti as Scientific Coordinator, and S. Hrelia as Responsible of
Research Unit)
Durata progetto
2008-2010
Abstract del progetto
The pathogenesis of ageing related neurodegenerative diseases,
such as Alzheimer's (AD) or Parkinson's (PD), is multifactorial with
the existence of a "domino" cascade of neurotoxic events, which
can be initiated at any point in the cascade. Many lines of evidence
suggest that oxidative stress, resulting in ROS/RNS generation,
mitochondrial dysfunction and activation of death/survival
pathways play a pivotal role in the age-associated neuronal loss in
neurodegenerative diseases. Thus, promising future treatment of
neurodegenerative diseases depends on availability of effective
brain permeable, antioxidant neuroprotective drugs that would
prevent the progression of neurodegeneration. An innovative
approach to preventive/therapeutic intervention in oxidative stress
based neurodegeneration may be the manipulation of endogenous
cellular defence mechanisms by chemical inducers, such as
isothiocyanates from natural sources, among which sulforaphane
(SF). Molecular and cellular mechanisms behind SF neuroprotection
will be initially studied in vitro, through an experimental approach
which let us to evaluate neuroprotective and/or neurorescue effects
by SF. Neuronal damage parameters at the cellular level will
comprise the induction of neuronal death in terms of
apoptosis/necrosis and the activation of cellular death/survival
biochemical pathways. The scientifically important expectation of
this project, by providing data on the cellular and molecular
mechanisms commanding cell death in neurodegeneration and
behind neuroprotection by SF, is to provide a clear rationale for
delineating innovative pharmacological strategies to counteract and
rescue neurodegeneration.
Fondazione IRCCS Istituto Neurologico “Carlo Besta”
Division of Neuropathology – Neurology 5
Director: Fabrizio Tagliavini
The research group
The Division comprises (i) a Clinical Unit (Dementia Center) devoted to the diagnosis and treatment of
patients with degenerative dementias (over 2000 out-patients and 150 in-patients per year) and (ii) a
Laboratory Unit dedicated to the analysis of genes and biomarkers associated with degenerative
dementias, post-mortem characterization of the disease process and disease-specific protein, and
experimental studies on disease pathogenesis and the development of therapeutic strategies. The course
of action of this activity can be expressed as a “Bed to Bench to Bed” cycle, with the ultimate goal to
identify disease-modifying drugs in preclinical settings and apply them to patients. In this regard, the
Clinical Unit is currently coordinating a multicentre phase II clinical trial supported by AIFA, to test the
effectiveness of an anti-amyloidogenic molecule identified in experimental models. This comprehensive
approach has also the great advantage to collect biological samples (plasma, DNA, CSF and brain tissue)
from fully characterized patients. In this regard the Laboratory Unit is part of the Network of Exellence
“BrainNet” Europe devoted to biobanking, harmonization of assessment tools and standardization of
diagnostic criteria of neurodegenerative diseases.
The permanent staff of the Division is composed by 8 MD and two technicians. In addition, the staff
comprises 13 post-doctoral fellows (7 PhD, 3 MDV, 1 MD, 1 psychologist), 2 PhD students and 1 technician
who are committed to the research activities on degenerative dementias with different expertise and roles.
On the overall, the team has large experience in a variety of techniques spanning from neuropathology
(including immunohistochemistry, morphometry, electron microscopy and atomic force microscopy), to
molecular genetics, biochemistry (purification and characterization of disease-specific proteins), cellular
and molecular biology, and animal models.
Research lines on Alzheimer’s disease
A. Clinical research
• Early diagnosis of Alzheimer’s disease (AD)
We will characterize the phenotypic expression of AD and other degenerative dementias through
multiplex analysis of a number of variables including neuropsychological and behavioural profile,
neuroimaging, neurophysiological changes with special attention to sleep abnormalities, genetics,
and plasma and CSF biomarkers. Furthermore, we will carry out longitudinal studies of
presymptomatic carriers of gene mutations to detect early markers of conversion to the disease
state.
• Clinical trials
We will carry out phase II and phase III clinical trials to assess the effectiveness of potentially
disease-modifying drugs, including molecules targeted to Aβ and protein tau.
• Identification of disease-specific biomarkers
We are currently working on a peripheral marker of AD having high specificity and sensitivity,
based on cyclic amplification of misfolded Aβ and Aβ oligomers from biological fluids, similar to
the PMCA technique successfully developed by Claudio Soto for prion diseases. The same
approach will be used to amplify and detect misfolded tau.
• Biobanking and neuropathological and molecular characterization of patients
We will collect systematically plasma, DNA and CSF from patients with different types of
dementia and non-demented individuals, and perform autopsies for neuropathological
characterization and biochemical and molecular studies.
B. Preclinical research
• Recessive A673V APP mutation: molecular mechanisms and development of a new therapeutic
strategy for sporadic AD
We have recently identified an APP mutation (A673V) that causes early-onset AD only in the homozygous
state while the heterozygous carriers are not affected. This mutation strongly boosts the production and
amyloidogenic properties of Aβ. However, the interaction of A673V-mutated and wild-type peptides
inhibits amyloidogenesis and Aβ-mediated neurotoxicity (Di Fede et al., Science 2009, 323:1473-7). These
findings are consistent with the observation that the A673V heterozygous carriers do not develop disease
and offer grounds for a novel therapeutic strategy based on modified Aβ peptides.
A research priority of our lab is to unravel the molecular mechanisms of the opposite effects of the
A673V APP mutation in homo- or heterozygous state on amyloidogenesis and to develop a lead
compound for AD therapy based on A673V-modified Aβ peptides or peptido-mimetic molecules.
To accomplish these objectives we have generated a panel of transfected cells and transgenic C.
elegans expressing human APP or Aβ with the A673V mutation, respectively, and transgenic
mouse lines expressing A673V-mutated APP in the homozygous or heterozygous state.
Furthermore, we have identified a prototypic lead compound corresponding to a six-mer Aβ peptide
with the A673V substitution and are currently working on brain delivery systems.
•
Pathways leading to tau pathology
A key event in AD pathogenesis is the hyperphosphorylation, misfolding and aggregation of the
microtubule-associated protein tau leading to formation of neurofibrillary tangles, disruption of the
neuronal cytoskeleton and neurodegeneration. Following the “Aβ cascade hypothesis” this event is
triggered by aggregated Aβ species, particularly oligomeric assembly intermediates. However, the
pathways linking Aβ and tau pathology are unknown. This is largely due to lack of animal models able to
reproduce these two central aspects of AD. We have developed a mouse model of prion disease that shows
a secondary tauopathy following deposition of PrP amyloid (Giaccone et al., Neurobiol. Aging 2008,
29:1864-73). This model will be used to investigate the molecular basis of tau pathology induced by
deposition of an amyloid protein.
• Role of nuclear tau in neurodegeneration in fronto-temporal dementia
Tau is the major microtubule-associated protein of neurons and its primary role is to promote assembly and
stabilization of microtubules required for morphogenesis and axonal transport. We have recently found that
tau plays an important role also in chromosome stability, and mutations in the tau gene cause chromosome
aberrations in peripheral cells in addition to formation of neurofibrillary tangles in neurons and glial cells.
Our objective is to elucidate the molecular mechanisms underlying the genomic instability due to tau
mutations using cellular models, and to investigate the contribution of the aneuploidy caused by mutated tau
to neurodegeneration using a transgenic mouse model of tauopathy.
Relevant ongoing research support
• Cariplo Foundation (01/01/07-30/06/10)
Title: Genoproteomics of Age Related Disorders (GuARD).
PI: Roberto Sitia
Role: Project leader
• Italian Ministry of Health (01/01/09-30/06/11)
Title: Alzheimer's disease: development and validation of a multi-factorial protocol for the diagnosis and
follow-up of disease in the prodromal and incipient phase.
PI: Fabrizio Tagliavini
• European Union/Italian Ministry of Health (01/02/09-31/01/12)
Title: Molecular mechanisms underlying synaptic dysfunction in prototypic neurodegenerative diseases
related to protein misfolding (nEUROsyn).
Role: PI
• Italian Agency of Drug (01/09/06-31/08/10)
Title: A randomized, double-blind pilot study versus placebo for the evaluation of the efficacy of
doxycycline administered by oral route in patients affected by Creutzfeld-Jakob disease.
Role: PI
Nome
Roberta Ricciarelli, Ph.D.
Contatti
E-mail: [email protected]
Tel.: +39-010-3538838; Fax: +39-010-3538836
Istituto/Dipartimento
Section of General Pathology
Department of Experimental Medicine (DiMeS)
University of Genoa, Italy
Proposta di ricerca
Dr. Roberta Ricciarelli’s ongoing research on Alzheimer’s disease:
•
Cholesterol and amyloid-beta: is there a relation?
•
Study of the nonmuscle myosin II involvement in the processing of the amyloid precursor protein (APP)
•
Specific type 4 phosphodiesterase inhibitors and their impact on the production of amyloid-beta
Area di interesse identificata
-Basic research,
-Multidisciplinary projects,
-New treatment strategies.
Finanziamenti ricevuti
Titolo progetto
Molecular mechanisms of N-truncated amyloid beta peptides
generation
Ente finanziatore
Durata progetto
Abstract del progetto
MIUR (Ministry of Education, University and Research)
Two years (2004-2005)
The amyloid precursor protein (APP) is an integral membrane protein
processed by several different proteases called secretases. β-Secretase is
responsible of the N-terminal cleavage of amyloid- (Aβ). The gene for βsecretase (also referred to as BACE) is located on chromosome 11, but no
AD-related mutation in this gene has been identified so far. BACE mRNA is
highly expressed in the brain and is also found in a variety of human
tissues, consistent with the fact that Aβ is normally produced by many
cell types. Recently, three new alternatively spliced transcripts of human
BACE have been described, but the functional importance of the different
BACE isoforms, and their implication in the pathogenesis of AD, has not
yet been investigated. Our preliminary sequence analysis indicates that,
as a cause of the alternative splicing, the DTG sequence, which is
conserved at the active site of the enzyme, may shift to a different
position within the catalytic site, affecting its substrate specificity. Since
BACE is involved in the generation of the N-terminus of Aβ species, the
existence of four BACE isoenzymes prompted us to investigate the
possible correlation between the expression of selective BACE isoforms
and the formation of neurotoxic N-truncated Aβ peptides. This
hypothesis is in line with the reported absence of AD-causing mutation in
BACE gene.
Titolo progetto
In vitro effect of PPAR-γ2 Pro12Ala polymorphism on the deposition of
Alzheimer's amyloid-β peptides
Ente finanziatore
Durata progetto
Abstract del progetto
Fondazione CARIGE
One year (2007)
Mounting evidence suggests that peroxisome proliferator-activated
receptor-γ (PPAR-γ) is involved in the modulation of pathogenic events
related to Alzheimer's disease (AD). Such events would include the
cerebral deposition of amyloid-β (Aβ) and the consequent local
inflammatory response. PPAR-γ has been shown to act on both fronts,
reducing either the secretion of Aβ or the expression of pro-inflammatory
cytokines. Recently, the relatively common Pro12Ala polymorphism in
exon 2 of PPAR-γ has been associated with higher risk for late onset AD.
The aim of the present project is to assess the impact of PPAR-γ and its
genetic variant on the secretion of Aβ in neuronal cultured cells.
Titolo progetto
Study of physiological function of the amyloid precursor protein
Ente finanziatore
Durata progetto
Abstract del progetto
University of Genoa
One year (2008)
The Alzheimer’s disease (AD) brain pathology is characterized by
extracellular deposits of amyloid- (A) peptides and intraneuronal
fibrillar structures. These pathological features may be functionally
linked, but the mechanism by which A accumulation relates to
neuronal degeneration is still poorly understood. A peptides are
fragments cleaved from the amyloid precursor protein (APP), a
transmembrane protein ubiquitously expressed in the nervous system.
Although the proteolytic processing of APP has been implicated in AD, the
physiological function of APP and the subcellular site of APP cleavages
remain unknown. The overall structure of the protein and its fast
anterograde transport along the axon support the idea that APP functions
as a vesicular receptor for cytoskeletal motor proteins. In the current
study, we will test the hypothesis that myosin II, important contributor to
the cytoskeleton of neuronal cells, may influence the trafficking and/or
the processing of APP. Our preliminary results demonstrated that
downregulation of myosin II-B, the major myosin isoform in neurons, is
able to increase A deposition, concomitantly altering the subcellular
localization of APP. These new insights might be important for the
understanding of the function of APP and provide a novel conceptual
framework in which to analyze its pathological role.
Nome
Contatti
Istituto/Dipartimento
Patrizia Guarneri
[email protected]
Istituto di Biomedicina e Immunologia Molecolare – Dip- di Medicina CNR- Palermo
Proposta di ricerca
Rare neurodegenerative diseases: basic research
Area di interesse identificata Rare diseases
Finanziamenti ricevuti
Titolo progetto
Pathogenesis of the CLN8 form of the late-infantile neuronal ceroid
lipofuscinosis
Ente finanziatore
1) MIUR “PRIN” Programmi di Ricerca Scientifica di Rilevante Interesse
Nazionale (DM n. 1407 del 4 dicembre 2008). “IDENTIFICATION OF
BIOLOGICAL, GENETIC AND CLINICAL MODIFIERS OF THE RATE OF
PROGRESSION AND SURVIVAL IN AMYOTROPHIC LATERAL SCLEROSIS ”,
prot. 20082XH2Z4-001
Durata progetto
Abstract del progetto
2) FIRB-MIUR, n. RBAU01E3SL, “RETINAL DEGENERATION IN MND
MOUSE, AN ANIMAL MODEL OF NEURONAL CEROID LIPOFUSCINOSES OR
BATTEN’S DISEASE. (dec. n. 808/Ric)
1) Two years
2) Three years
The neuronal ceroid lipofuscinoses (NCLs) are a group of
neurodegenerative, lysosomal diseases with onset usually in childhood,
being characterized by progressive visual loss, dementia, motor decline,
therapy resistant epilepsy and early death. Our team has lately focused
on the pathogenic mechanisms of the CLN8 variant form of the lateinfantile NCLs (vLNCL), which is present in Finland, Turkey and Italy. The
CLN8 gene encodes a transmembrane protein which is located mainly in
the ER and partially in the ER-Golgi intermediate compartment of both
neuronal and non-neuronal cells, and it is supposed to have a role in the
ceramide synthesis. However, its exact function remains undiscovered.
We have developed a protein-protein interaction study throughout the
screening of a human brain cDNA library with split-ubiquitin membrane
two-hybrid system in yeast, which is the only technology developed thus
far that can work effectively as a in vivo screening system to find
interactors of ER membrane proteins. This approach along with
coimmuno-precipitation and -localization assays of transfected cells has
envisaged for the first time four major protein interactors with the CLN8
protein. Through in vitro and in vivo studies, we are looking at the
definition of the functional role of these interactions and particularly of
the CLN8 interaction with two of the characterized proteins, which are
known to have functions in synthesis and transport of lipids and are
suggested to be involved in the amyothrophic lateral sclerosis. In an
attempt to understand the mechanisms leading to neuronal cell death in
the disease, we have been also using a spontaneous animal model of the
CLN8/vNCL, the mnd mouse. We are currently investigating whether and
how dysfunctions of ER/UPR system and autophagolysosomal pathway
cooperate in the neurodegeneration of the retina and brain structures of
the mnd model.
Nome
Dario Finazzi
Contatti
[email protected]
phone and fax ++39 030 302356
Dip. Materno Infantile e Tecnologie Biomediche
Università degli studi di Brescia
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
1) Basic research
2) Genetic susceptibility to Alzheimer’s disease and genome wide
association studies
3)
Biobanking
Finanziamenti ricevuti
Titolo progetto
1) IRON METABOLISM IN NEURAL CELLS AND IN
NEURODEGENERATIVE DISORDERS
Ente finanziatore
2) ANALYSIS OF CANDIDATE GENES OF SUSCEPTIBILITY TO AD.
University of Brescia
Fondazione RSA Agostoni Lissone
Durata progetto
3 years
Abstract del progetto
Our group is interested in the study of iron metabolism in the brain and
its possible role in neurodegeneration. The accumulation of iron is a
phenomenon often detected in the brain from patients affected by
Alzheimer’s disease and by other neurodegenerative processes. Even
though such accumulation is not the primary cause of neural death, it is
surely involved in the progression and deterioration of the
neurodegenerative process. The excess of iron can be directly linked to an
increase in the production of free radicals, leading to oxidative damage to
proteins and membranes, signs commonly documented in tissues with
neurodegeneration. We aim to a better comprehension of such processes
by developing two different and complementary lines of research: 1)
Analysis of the biological mechanisms involved in the maintenance of iron
homeostasis in the brain, in normal and pathological conditions; 2)
Analysis of genes linked to iron metabolism as possible modifiers of the
individual risk to develop AD.
We intend to perform a thorough analysis of the regulation of iron
metabolism in neural cells and to assess the way its malfunction may lead
to iron accumulation. The experiments will be initially carried out in SH5YSY cells (either differentiated or not) and in primary hippocampal
neurons with the goal to characterize the role played by the hepcidinferroportin axis in iron homeostasis of neural cells. We will determine the
mechanism regulating ferroportin expression and function and describe
the molecular machinery controlling hepcidin production in neuronal
cells and the role of the peptide in neuronal iron trafficking. The analysis
will be then extended to wild type mice and murine model of AD
(APP/PS1dE9), and eventually to autopsy sections of human brain from
patients and controls. In particular we will investigate the relationship
between brain inflammation and the regulation of the hepcidinferroportin axis. We have preliminary evidence that BMP6 and
Interleukins intervene in the transcriptional regulation of hepcidin and
could be of relevance in the progressive accumulation of iron in brain.
The study take advantage of a significant collaboration with Prof Paolo
Arosio, one of the major expert in the study of cellular iron metabolism.
The genetic part of the project will investigate the possible association
between polymorphisms in genes involved in iron homeostasis (hepcidin,
ferroportin, DMT1, ceruloplasmin) and the individual risk to develop AD.
Promoter regions, exons and exon-intron boundary regions will be
scanned for the presence of sequence variations by DHPLC and real time
genotyping. The study is granted by the availability of a large set of
genomic DNA samples from patients and controls and by the
collaboration with research groups and clinicians with expertise in AD and
genetic studies (prof. Scarpini, Dott. Galimberti, Dott. Dominici). In 2005
we set up a bank of different biological samples coming from AD patients
and sex and age matched controls. The biobanking activity is continuing
thanks to the collaboration with the Fondazione RSA Agostoni in Lissone
and Dr Roberto Dominici. Our valid collaboration with the group of Prof
Scarpini at Ospedale Maggiore in Milan allows us the access to a large and
well characterized set of genomic DNA samples, as a second and different
population for the confirmation of data obtained in the first set of
patients and controls.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Proposta di ricerca
Carlo Sala
Carlo Sala
+390250317096
[email protected]
CNR Istituto di Neuroscienze
Area di interesse identificata
Neurotoxic stimuli, synaptic dysfunction
Finanziamenti ricevuti
Titolo progetto
The role of intraneuronal amyloid β(Aβ) peptide on the pathogenesis of
the Alzheimer disease: functional and proteomic analysis.
Ente finanziatore
Fondazione CARIPLO
Durata progetto
2 anni
Abstract del progetto
La malattia di Alzheimer rappresenta la più comune forma di demenza
per la quale non esiste ancora nessun intervento terapeutico in grado di
prevenire, curare o ritardare in modo sostanzaile il suo sviluppo. Recenti
studi hanno dimostaro che l’accumulo intracellulare di Abeta correla con
la prima comparsa di deficit cognitivi. Questo progetto si pone
l’obbiettivo di identificare come Abeta intrecelluare interfereisce con la
funzionalità delle sianpsi neuronali e, con tecniche proteomiche avanzate,
di individuare le proteine che interagiasono con Abeta. Infine ci
proponiamo di studiare i meccanismi molecolari alla base della
formazione di Abeta purificando e caratterizzando la composizione
proteica delle vescicole di trasporto per APP. I risultati che ci proponiamo
di ottenre con questo porgetto potranno contribuire alla conoscenza dei
meccanismi patogenetici dell’AD e potrebbero consentire lo sviluppo di
nuovi metodi diagnostici e la eventuale messa a punto di protocolli
terapeutici
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Cristina Limatola
+39 06 49690243 +39 347 2924450
[email protected]
Dipartimento di Fisiologia e Farmacologia, Università Sapienza Roma
Tra le altre linee di ricerca di interesse del laboratorio, siamo da qualche
anno coinvolti in studi di ricerca sulla sclerosi laterale amiotrofica (SLA).
In particolare, in collaborazione con un gruppo di neurologi (Sabatelli,
Neri) e genetisti (Zollino) dell’Università Cattolica di Roma, abbiamo
descritto, in pazienti affetti da SLA, delle mutazioni a carico di diverse sub
unità del recettore nicotinico neuronale, nel loop regolatorio
citoplasmatico (Sabatelli et al., 2009). Queste mutazioni, in buona misura
a carico di potenziali siti di fosforilazione da parte di protein chinasi,
determinano delle alterazione funzionali specifiche di alcune sub unità
per recettori-canale per l’acetilcolina, con una generale “gain of function”
recettoriale. L’aumentata responsività alla stimolazione con acetilcolina a
livello dei terminali glutamatergici potrebbe determinare una liberazione
eccessiva di glutammato eccito tossico, mentre la potenziata attività dei
recettori nicotinici potrebbe aumentare l’ingresso di calcio intraneuronale
a livelli tossici, contribuendo all’inizio o alla progressione della malattia.
Stiamo attualmente continuando questi studi molecolari, analizzando gli
effetti dell’adenosina e dell’ATP, i cui livelli sono alterati nella risposta
infiammatoria cronica caratteristica della SLA, sull’attività dei recettori
nicotinici neuronali mutati individuati rispetto alla controparte normale.
Altro punto di interesse riguarda il coinvolgimento delle cellule gliali,
astrociti e microglia, nello sviluppo e nella progressione di questa
patologia. Abbiamo sviluppato a tal proposito dei sistemi di co-coltura
motoneuroni-cellule gliali e di fettine ottenute dal tronco cerebrale
(regione del nucleo ipoglosso) per studiare il contributo della
componente gliale nella alterazione dell’attività elettrica ritmica, nelle
proprietà dei recettori per neurotrasmettitori, sensibilità
all’eccitotossicità da glutammato, e ci proponiamo di applicare questi
studi a modelli cellulari che overesprimano forme mutate di proteine
associate alla SLA, come FUS, TDP-43 o SOD (in collaborazione con un
gruppo di biologi molecolari della nostra università, Prof. Bozzoni).
Studio funzionale di mutazioni che aumentano la suscettibilità alla
Sclerosi Laterale Amiotrofica su calciatori amatoriali o professionisti.
Fondazione Milan
2010-2011
La sclerosi laterale amiotrofica (SLA) è una malattia devastante che
distrugge in maniera selettiva una particolare categoria di cellule nervose:
i motoneuroni. Questi sono i neuroni che controllano la contrazione dei
muscoli scheletrici, per cui la loro morte provoca una paralisi progressiva
dei soggetti affetti da SLA e, nel giro di 2-6 anni dall’insorgenza della
malattia, la loro morte (solitamente per arresto respiratorio).
Solo il 5% dei casi di SLA ha chiaramente un’origine ereditaria, nel
restante 95% la malattia ha un origine incerta (SLAs). Tra i giocatori di
calcio, caso unico tra tutte le categorie di sportivi professionisti, il rischio
di sviluppare una SLAs è ben 6 volte superiore a quello della popolazione
generale. Non è ad oggi nota la ragione di questo fenomeno. Si ritiene
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
che i principali fattori di rischio per lo sviluppo delle SLAs, oltre all’età e al
sesso maschile, siano i traumi meccanici, l’esposizione ad erbicidi,
un’attività fisica molto intensa e il fumo di sigarette. Da un punto di vista
cellulare, una delle cause che si ritiene essere alla base di questa malattia,
è il danno prodotto dal massiccio aumento dello ione calcio all’interno del
neurone. Siccome la stimolazione dei recettori della nicotina (la sostanza
contenuta nelle sigarette) che sono localizzati sulla membrana dei
motoneuroni consente l’ingresso del calcio all’interno del motoneurone si
è pensato di studiare la frequenza delle mutazioni dei geni che codificano
i recettori nicotinici neuronali. Queste mutazioni sono presenti nel 6% di
pazienti affetti da SLAs e inducono tutte un’alterazione nel
funzionamento dei recettori nicotinici neuronali che provoca un aumento
dell’ingresso del calcio all’interno del motoneurone aumentando la
probabilità che esso muoia. Data l’anormale frequenza di questa malattia
nelle persone che praticano l’attività calcistica a livello agonistico
(amatoriale o professionale), ci proponiamo di valutare, mediante analisi
del DNA estratto con semplici prelievi di sangue, la presenza di mutazioni
dei recettori nicotinici in calciatori in buone condizioni di salute e/o di
coloro che si sono già ammalati di SLAs. Siamo convinti che lo studio delle
modalità precise delle modificazioni del funzionamento dei recettori
nicotinici neuronali dovute a queste mutazioni genetiche, possa essere il
primo e fondamentale passo per sviluppare una terapia innovativa mirata
a ripristinare il corretto funzionamento dei recettori con la accelerazione
della loro desensibilizzazione, diminuendo significativamente il rischio
dell’insorgenza della malattia, cioè la suscettibilità alla malattia. A questo
scopo ci proponiamo di caratterizzare dal punto di vista funzionale le
mutazioni trovate in sistemi eterologhi ricostituiti in vitro (linee cellulari
trasfettate); inoltre intendiamo sviluppare un modello di topo
transgenico per una delle mutazione dei recettori nicotinici associata a
SLA, e utilizzarlo come modello in vivo per studiare la fisiologia del
muscolo e la fisiologia neuronale.
Molecular and functional approaches to investigate the
physiopathological role of the chemokines and their receptors in the
central nervous system.
Fondazione Cenci Bolognetti
2007-2010
Chemokines and their receptors are widely expresses in CNS where they
play important roles both under physiological and pathological
conditions. Chemokines are implicated in the control of neuronal and glial
development, both in terms of migration and maturation, modulate
synaptic transmission and have trophic and protective effects on neurons
and glia. The spectrum of biological activities of chemokines is broadened
by their involvement in the pathogenesis and maintaining of several
neuropathologies, when their expression may be altered. Aim of the
proposed research project is to investigate the physiopathological
activities of chemokines as modulators of neuronal functions, identifying
the molecular pathways involved. In particular the effects of chemokines
will be investigated in brain slices, primary neuronal cultures, cell lines
and expression systems as concern synaptic transmission, short-term
neuromodulatory activities and long-term plasticity, trying to correlate
these events to the activation of specific molecular effectors. These
investigations will be correlated with studies of chemokine-induced
communication between different cell types, like glial cells and neurons.
The effects of chemokines as modulators of cell migration will be
investigated in glial cells and neurons freshly isolated from rodents and
the signal transduction pathways involved will be investigated. The
activities of chemokines as neuroprotective substances will be
investigated in vitro in neurotoxicity models and in vivo in animal models
of cerebral ischemia. Overall, this study aspire to help to improve the
understanding of neurogenesis and neurorepair, yielding possible clues to
therapeutical strategies.
IIT DEPARTMENT OF NEUROSCIENCE AND BRAIN TECHNOLOGIES
ONGOING PROJECTS ON NEURODEGENERATION
Fabio Benfenati, MD
1. Exposing tau pathology at its earliest
Team: Laura Gasparini; Axel Blau, Paolo Medini; in collaboration with M. G. Spillantini, M. Goedert
Funding: Compagnia di San Paolo and IIT + pending.
The study is aimed at revealing functional changes typical of initial stages of tauopathy, and elucidating
the underlying molecular mechanisms. This knowledge will reveal markers suitable for early clinical
detection of tau-driven degeneration and identify key pathogenetic mechanisms to be targeted by new
therapies.
2. Effects of β-amyloid aggregates on neuronal viability and microglial function
Team: Denise Ferrera, Claudio Canale, Fabio Benfenati, Laura Gasparini
Funding: IIT + pending
We have demonstrated that Ab42, but not Ab40, reproducibly aggregates into stable oligomers and
protofibrils and reduces the viability of mouse hippocampal neurons. We are now investigating how
distinct Aβ aggregates affect microglia viability and activate microglial neuroinflammatory response
and the latter response on neuronal physiology.
3. β-amyloid aggregates and astrocytic function in Alzheimer pathophysiology
Team: Tommaso Fellin, Laura Gasparini, Nadia Mazzaro
Funding: IIT + pending
We will use an interdisciplinary approach including two-photon microscopy, Ca2+ imaging,
biochemistry and pharmacology to test the hypothesis that b-amyloid (Ab) oligomers induce reactive
gliosis through the overactivation of astrocytic NMDA receptors and we will evaluate whether the
NMDA open-channel blocker memantine antagonizes Ab-induced astrocytic reactive response.
4. Molecular and structural determinants for β-amyloid mitochondrial accumulation and toxicity
Team: Mounia Chami, Fabio Benfenati, Dolores Del Prete
Funding: IIT + pending
Mitochondrial dysfunction is emerging to play a pivotal role in Alzheimer’s disease (AD). Our project is
aimed at: (i) studying ER-mitochondria communication and APP and Aβ distribution in AD models; (ii)
modulating ER-mitochondria communication by pharmacological (Ca2+-drugs) and genetic/molecular
biology through expression of mitochondrial fission and fusion proteins and/or S1T and consequently
analyse mitochondrial function, and the intracellular distribution of APP and Aβ; (iii) determining the
APP/Aβ partners in the MAM fraction using a proteomic approach.
5. Calcium modulating drugs as a therapeutic tool for Alzheimer’s disease
Team: Mounia Chami, Fabio Benfenati, Dolores Del Prete
Funding: IIT + pending
The ability of neurons to regulate Ca2+ signals is compromised during aging. Importantly, alteration of
Ca2+ regulating systems has been shown to occur early in Alzheimer’s disease (AD). We will
investigate the potential use of Ca2+ drugs as a mean to reduce Aβ production in different AD models
in vitro, analyze the potential effect of Ca2+ drugs on beta and gamma secretase activity and apply Ca2+
drugs on animal models in vivo and consequently analyze their effect on Aβ plaque formation, synaptic
activity and cognitive development and/or progression.
6. Pathogenesis and therapeutic targets for SBMA
Team: Mara Pennuto, Fabio Benfenati, Chiara Scaramuzzino
Funding: MDA, Marie Curie, IIT + pending
Polyglutamine diseases belong to the broad family of brain folding diseases. We aim at using Spinobulbar
muscular atrophy (SBMA) as a model of polyglutamine disease to identify signaling pathways
that impact brain folding diseases. Recent evidence shows that the IGF-1/Akt signaling pathway impact
brain folding diseases. However, the mechanisms underlying the effect of Akt signalling remain to be
elucidated. By putting polyglutamine AR downstream of Akt with our previous research, we
established SBMA as a good model to tease apart the molecular details of the effect of IGF-1/Akt in
polyglutamine disease.
7. Interactions of Aβ and α-synuclein with the cytoskeleton
Team: Evelina Chieregatti, Giuseppe Ronzitti, Anako Tsushima
Funding: IIT, Cariplo Foundation + pending
Both Aβ and α-synuclein are implicated in actin dynamics as well as microtubule stability through
their interactions with a microtubule-binding protein, tau. Our projects concern: (i) the study of their
effects on the morphology and dynamics of actin microfilaments and intermediate filaments; (ii) the
possible effect of the two proteins on cytoskeletal-regulated processes, such as polarization, growth
cone formation and regeneration after axotomy; (iii) the identification of proteins of interest such as
enzymes or molecular motors that may be found in the APP-containing endocytic vesicles.
8. Mechanisms of neurodegeneration and enhancement of adult neurogenesis to improve
cognitive functions in Down syndrome
Team: Andrea Contestabile, Diego Ghezzi, Fabio Benfenati, Laura Gasparini
Funding: Lejeune Foundation + IIT + pending
Down syndrome (DS) patients develop neuropathological lesions typical of Alzheimer disease (AD) by
the fourth decade of life with cerebral accumulation of amyloid-beta (Ab) deposits and intraneuronal
inclusions made of hyperphosphorylated tau protein (neurofibrillary tangles, NFTs). Mice models of
DS recapitulate many aspects of the pathology, including: APP-dependent cholinergic dysfunctions
during aging and tau hyperphosphorylation. This research line aims to better characterize the
neurodegeneration process driven by APP overexpression and tau hyperphosphorylation in a DS mouse
model and evaluate potential therapeutic interventions to alleviate the pathology. Down syndrome is
also characterized by decreased neurogenesis both during development and adulthood. Adult
neurogenesis in the DG of rodents has been associated to memory formation and hippocampaldependent
learning. This research line aims to elucidate the impact of adult neurogenesis on learning
and memory in a mouse model of DS and assess the effect of pharmacological treatments able to
enhance the neurogenesis process.
Nome
Armando Genazzani
Contatti
Tel.
E mail
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
DiSCAFF, Via Bovio 6 - Università del Piemonte Orientale, Novara
0321-375827
[email protected]
DiSCAFF, Università del Piemonte Orientale
Ente finanziatore
Durata progetto
Abstract del progetto
Malattia di Alzheimer e trasduzione del segnale legata al calcio
Role of calcium-dependent gene expression and protein trafficking in
shaping post-synaptic activity: implications for neurodegenerative
diseases
Fondazione Cariplo
2 anni
Alzheimer disease (AD) is a progressive neurodegenerative disease
characterized by a progressive cognitive and memory decline, increasingly
frequent with advancing age until it affects as many as 50% of individuals
85 years of age and older. There is no remission in the progression of the
disease nor are there any truly effective pharmacological interventions
available at present. In the framework of an ageing society a better
understanding of pathogenic mechanisms and identification of new
pharmacological targets are key strategies.
For many years, AD was attributed to neuronal death induced by deposits
of fibrillar Amyloid b. The production of amyloid ß (Aß) is mediated by the
concerted action of two different secretases, namely ß-secretase (BACE)
and gamma-secretase, showing a proteolytic action on the amyloid
precursor protein (APP). Alternatively, APP can be subjected to the
proteolytic cleavage by a--secretase (ADAM10), which occurs within the
sequence of Aß, thus precluding the formation of the amyloidogenic
fragments. Yet, a recent view suggests that Abeta production has an
impact primarily on the post synaptic compartment of the excitatory
glutamatergic synapse (this preceding cell death and correlating with
clinical features of the disease), as demonstrated by in vitro studies, thus
indicating Alzheimer disease as a synaptopathy.
An additional biochemical parameter which has been linked to both
synaptic plasticity events and neurodegeneration is Calcium. The link
between calcium and Alzheimer has been strengthened by a number of
independent groups. Calcium dyshomeostasis appears to be an integral
part of the pathology, either because it can influence Abeta production or
because it can be altered by amyloid beta overproduction.
This project will focus primarily in dissecting the contribution of calcium
dyshomeostasis to the AD synaptopathy. Thus, the main goal of the
project is to investigate the link between dys-regulation of calcium
signalling and early stages of AD – both at cellular and molecular level working on the hypothesis that this neurodegenerative disorder is a
synaptopathy. In particular we will firstly characterize calcium dependent
processes in an innovative animal model of Alzheimer disease obtained
by uncoupling ADAM10 from its cargo protein SAP97 which regulates
enzyme activity. Secondly, we will address the role of calcium in both
trafficking and gene expression of key synaptic proteins in vitro and in in
vivo models of altered amyloid cascade.
The investigation of these objectives would provide novel insights in the
understanding of the molecular basis of Alzheimer’s disease in early
stages and would possibly contribute in identifying new targets for
therapeutic intervention
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse
identificata
Dr. Andrea Contestabile
Email: [email protected]
Phone: ++39-010-71781785
FAX:
++39-010-71781230
The Italian Institute of Technology (IIT)
Department of Neuroscience and Brain Technologies
Via Morego 30, 16163 Genova, Italy
Down syndrome (DS) or trisomy 21 is the leading cause of genetically-defined
mental retardation and is characterized by decreased cognitive functions,
developmental defects, impaired neurogenesis and Alzheimer (AD)-like
neurodegeneration during aging. The progression of Alzheimer neuropathology in
DS individuals appears to be analogous to sporadic cases of AD with cerebral
accumulation of amyloid-beta (A) deposits (i.e., A plaques) and intraneuronal
inclusions made of hyperphosphorylated tau protein (i.e., neurofibrillary tangles,
NFTs). However, its appearance is anticipated by at least twenty-thirty years with
respect to the general population. The presence of an extra-copy of the
Aprecursor protein (APP) gene, located on human chromosome 21 (HSA21), is
believed to be directly responsible for early deposition of A in DS brain. The gene
encoding Dyrk1A is also located in the triplicated region of HSA21 and is
overexpressed in DS brains. Recently it has been proposed that Dyrk1A may be
involved in tau hyperphosphorylation and NFTs formation typical of DS and AD.
Dyrk1A phosphorylates tau at several sites, and such sites are
hyperphosphorylated in adult DS brains. In addition, phosphorylation by Dyrk1A
primes further phosphorylation of tau by Glycogen Synthase Kinase-3 (GSK-3
suggesting that the extra copy of the Dyrk1A gene on HSA21 contributes to the
early onset of AD neuropathology seen in DS. In fact, a role for Dyrk1A in AD
neurodegeneration has been recently identified, therefore suggesting a functional
link between some of the triplicated genes found in Down syndrome and Alzheimer
disease.
The Ts65Dn mouse is the best characterized animal model of DS and carries an
extra copy of the distal segment of mouse chromosome 16 which is synthenic to
human chromosome 21. The Ts65Dn mouse recapitulates many aspects of DS,
including: APP-dependent cholinergic dysfunctions during aging; Dyrk1A-mediated
tau hyperphosphorylation and impairment of hippocampus-dependent memory
function.
Ts65Dn mice also display reduced adult neurogenesis in the two known adult
neurogenic niche in mammalian brain, i.e, the subventricular zone (SVZ) of the
lateral ventricle and the subgranular zone (SGZ) of the dentate gyrus (DG) in the
hippocampus. Adult neurogenesis has been also found to be reduced in several
mouse models of AD, further supporting the notion of a functional connection
between DS and AD pathological mechanisms. Adult neurogenesis in the DG of
rodents has been associated to memory formation and hippocampal-dependent
learning such as spatial and contextual recognition. Decreased adult neurogenesis
observed in DS and AD mouse models is therefore believed to play an important
role in the impaired performance of these mice in hippocampus-depended
memory tasks.
GSK-3 inhibition has been identified has a possible therapeutic target for AD
neurophatology. Accumulating evidence also indicates that adult neurogenesis is
negatively regulated by the activity of GSK3 through the inhibition of the Catenin pathway, and that adult DG neurogenesis can be stimulated by GSK3
inhibitors.
In order to identify effective pharmacological treatment to improve learning and
memory in mouse models of DS that could be translated into human therapy for
both AD and DS we are currently evaluating the usefulness of GSK-3 inhibitors to
stimulate adult DG neurogenesis and alleviate tau hyperphosphorylation in Ts65Dn
mice.
Finanziamenti ricevuti
Titolo progetto
Effects of lithium on cognitive functions, neurogenesis and Tau pathology in
Ts65Dn mice.
Ente finanziatore
Fondation Jerome Lejeune, France
Durata progetto
2 years
Abstract del
Compelling studies indicate that impairment of adult neurogenesis and
progetto
degenerative mechanisms driven by hyperphosphorylation of Tau are the main
determinants in reducing cognitive and memory function in animal models of
Down syndrome (DS). Indeed, adult neurogenesis is impaired in the dentate gyrus
of the hippocampus in the Ts65Dn mouse, a DS model reproducing most
neuropathological and cognitive deficits of the disease. We have recently shown
that administration of lithium to Ts65Dn mice is able to restore neurogenesis in the
subventricular zone of the lateral ventricle, another neurogenetic area of the adult
brain. Lithium is an extensively used mood stabilizer and acts as an inhibitor of
Glycogen Synthase Kinase-3(GSK-3). GSK-3 activity enhances tau
phosphorylation and is primed by dual-specificity tyrosine phosphorylated and
regulated kinase 1A (Dyrk1A), which is encoded by a gene triplicated in DS and
Ts65Dn mice. Aim of the present project is to test whether lithium long-term
administration ameliorates cognitive performance in Ts65Dn mice by reducing
accumulation of hyperphosphorylated tau and by stimulating neurogenesis,
differentiation and integration of newly-born neurons. Lithium (2.4 g/kg of Li2CO3
in the chow) will be administered to Ts65Dn for 4 or 8 weeks starting at 6 months
of age. Hippocampus-dependent memory functions will be evaluated through
novel object recognition, spontaneous T-maze alternation and geometric task tests
performed at the end of lithium treatment. After the treatment, adult
neurogenesis in the dentate gyrus of Ts65Dn mice will be assessed by BrdU labeling
and evaluation of proliferation markers. The expression of neuronal differentiation
markers will be analyzed to assess the maturation of newly generated neurons. The
accumulation of hyperphosphorylated Tau will be quantitatively monitored by
immunohistochemical and biochemical techniques. Our experimental approach will
allow testing a novel therapeutic approach for DS by simultaneously targeting two
known pathological features affecting cognitive functions.
Nome
LAURA GASPARINI
Contatti
[email protected]
Istituto/Dipartimento
Fondazione Istituto Italiano di Tecnologia, Dipartimento di Neuroscienze
e Neurotecnologia, Via Morego 30, 16163 Genova
Proposta di ricerca
Area di interesse identificata
The lab focuses on studying key neuropathological molecular mechanisms
of neurodegenerative diseases, with the view of understanding the
disease processes and to translate basic knowledge into potential novel
therapies.
Tau-driven degeneration and neurotrophic response: searching disease
markers for drug screening. Accumulation of intraneuronal incusions
made of tau protein is a pathological hallmark of the brain of patients
with tauopathies, including frontotemporal dementia and parkinsonism
linked to chromosome 17, parasupranuclear palsy, corticobasal
degeneration and Alzheimer Disease (AD). Our research is aimed at
providing insight on alterations of neuronal activity associated with the
onset and progression of tau pathology in transgenic disease models of
tauopathy in vitro and in vivo, to reveal key tau-driven pathological
mechanisms, and identify potential molecular targets for drug discovery.
Neuroinflammatory mechanisms in Alzheimer disease: role of β-amyloid
aggregates in microglia activation. Neuroinflammation is one of the
neuropathological features of Alzheimer Disease (AD) and is characterized
by activation of inflammatory cells (i.e., astrocytes and microglia) in areas
of the brain affected by β-amyloid (Aβ) plaques and tau pathology, with
production of altered levels of inflammatory mediators. Our lab
investigate the pathogenic mechanisms underlying AD-associated
neuroinflammatory processes, with the main focus on understanding the
role on Aβ aggregated species in microglia activation.
Pathogenic mechanisms driven by laminB1 overexpression in adult
onset leukodystrophy. Lamin B1 (LMNB1) is a component of the nuclear
lamina. It has been recently discovered that the presence of an extra copy
of the gene encoding this protein causes autosomal dominant
leukodystrophy (ADLD). ADLD is an invariably fatal disease characterized
by white matter loss, movement disorders and dysautonomia. The
pathogenic mechanisms by which LMNB1 gene duplication leads to
disease are unknown. Our research aims at clarifying such mechanisms
using in vitro and in vivo experimental models.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Tau-driven degeneration and neurotrophic response: searching disease
markers for drug screening
Compagnia di San Paolo
3 years
Background: Neurofibrillary tangles (NFTs) are neuropathological
hallmarks of human tauopathies. Reduced levels of neurotrophin TrkB
receptor have been found in NFT-bearing neurons. Rationale and
objectives: Initial findings showed that retinal ganglion cells (RGCs) of
human P301S tau transgenic (TG) mice develop NFTs and have impaired
axon outgrowth upon stimulation with neurotrophins (CNTF/BDNF). We
propose to investigate the mechanisms by which tauopathy affects
neurotrophin signalling and neuronal activity to identify disease markers
and provide a test-bed for evaluating potential neuroprotective drugs in
vitro. Expected results and relevance: Reveal characteristic signatures of
tau pathology, suitable to investigate potential tau-targeting
therapeutics.
Titolo progetto
Ente finanziator
Durata progetto
Abstract del progetto
Clinical, neuroradiological and molecular investigation of Adult-onset
Autosomal Dominant LeukoDystrophy (ADLD): dissection of Lamin B1mediated pathophysiological mechanisms in cellular and mouse models.
TELETHON
3 years
Overall objectives: To clarify Lamin B1 (LMNB1)-mediated pathogenic
mechanisms of adult-onset autosomal dominant leukodystrophy (ADLD)
and provide tools to study future therapies. Background/Rationale: ADLD
is a rare, progressive and fatal genetic disease generally characterized by
autonomic failure at onset. We have collected two ADLD families with
LMNB1 gene duplication, and a third with a variant ADLD and an
unknown mutation affecting LMNB1 expression. Expected results: The
project will enhance our knowledge on genetic, clinical and
neuroradiological features of ADLD. It will shed light on its
pathophysiology and on LMNB1-mediated pathogenic mechanisms. It will
generate an ADLD mouse model and provide tools (i.e., clinical
biomarkers, expression constructs, gene-reporter assays, cellular models)
which will be key elements for further studies of the disease and its
therapy.
Nome
Contatti
Michele Mazzanti
[email protected]
+39 2 50314958 +39 2 50314959
Istituto/Dipartimento
Dept. of Bimolecular Science and Biotechnology
Proposta di ricerca Investigation on early markers during neurodegenerative process
Area di interesse identificata
D. Oxidative Stress and neurodegeneration
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Beta-amyloid effect on retina cells.
Investigation on Chloride Intracellular Channel 1 (CLIC1) protein modulation to use as an
early marker during neurodegenerative process induced by amyloid compounds.
Merz Biotech, Germany
Italian Research Ministery
1 year
2 years
One of the major apprehensions of contemporary society is population aging.
Paradoxically, scientific discoveries applied to medicine have significantly increased the
life expectation. However, older populations create new medical problems that, once
episodic, are now common in a growing number of aging individuals.
Neurodegenerative diseases play a predominant role in this new scenario. A major
neurodegeneration disorder is Alzheimer Disease (AD), but this is poorly understood
and not only we are without a successful therapy but also a proper identification of the
disease in living humans is, at the moment, problematic. The appropriate diagnosis of
an AD patient can be obtained only from histological analysis of post-mortem brain
tissue. AD is characterized by an accumulation in neuritic plaques of beta-amyloid (A)
protein, infiltrated by reactive microglia. Although A and in particular its oligomeric
forms has been shown to exert a toxic effect on neurons, the role of microglia in
neuronal injury remains unclear. Microglia activation is likely to be a key element in the
pathophysiology of the local inflammation that accompanies AD. Activation of microglia
by A protein triggers the production of toxic free radicals and pro inflammatory
cytokines which are responsible for the neuronal degeneration. Previous studies
demonstrated that A protein stimulation of neonatal rat microglia specifically lead to
the increase of Chloride Intracellular Channel 1 (CLIC1) expression after 48 hours of
stimulation. This was not the case when microglia was activated by phorbol myristate
acid (PMA), Lipopolysaccharide, colony stimulating factor or basic fibroblast growing
factor, thus quoting CLIC1 as specifically up-regulated by amyloid compounds. The
cellular and molecular characterization of the protein has revealed that CLIC1 is mainly
cytoplasmatic and is able to shuttle between the aqueous and the membrane phase
depending on the activation state of the cell. In the membrane, the protein acts as a
chloride conductance, allowing ionic flow to compensate the negative charges extruded
by the membrane NADPH oxidase during reactive oxygen species (ROS) production. We
have recently showed that suppression of CLIC1 functional expression using (i) inhibition
of the CLIC1 current, (ii) replacement of extracellular Cl with impermeant anions, (iii)
transfection of microglia cells with specific siRNA, or (iv) addition of an antibody against
the channel protein, significantly reduced Aβ-induced ROS production. In a triple
mutant Alzheimer mouse model, CLIC1 protein is up-regulated and show a predominant
membrane localization in microglia cells of cerebral cortex. Furthermore, in human
brains of Alzheimer patients, CLIC1, like several other proteins, is drastically up
regulated. To explore the idea that CLIC1 protein could work as a therapeutic target as
well as specific early marker for amyloid based diseases, we will investigate its
distribution and modulation in two other neurodegenerative processes. Prion Protein
syndrome (PrPsc) and Spinocerebellar Ataxia 1 (SCA-1) share with AD the predominant
-sheet structure of the presumed pathological protein. As typical amyloid compounds,
A, PrP and Ataxin-1 are characterized by insoluble deposits. However, while the action
mechanism of the first two takes place in the extracellular environment, Ataxin-1
accumulates in the cell nucleus.
The goal of our study is to pursue CLIC1 as a molecular target for early diagnosis and
possibly for a therapy of amyloid induced neurodegeneration. Since its specific reaction
to amyloid peptides, CLIC1 could represent an unambiguous marker of several amyloid
based neurodegenerative process. It is reasonable to think that its marked increase in
the brain immune system occurs in the early stage of the pathological process following
a chronic state of oxidative stress induced by the different amyloids accumulation. We
will use two different strategies to assess this possibility. It is well known that during
neurodegenerative states the central nervous system (CNS) is colonized by systemic
macrophages recruited from the blood stream. It is likely that the up regulation of CLIC1
protein could be a general feature of all macrophages. This will be tested both in mouse
models of AD, PrPsc and SCA-1 as well as in blood samples from patients. Alternatively,
we will explore the possibility to detect CLIC1 increment in the most accessible part of
the CNS: the retina. Activation of the CNS immune system and neurodegenerative
process were described in the eyes of AD, PrPsc and SCA-1 patients. Degeneration of
retinal ganglion cells (RGCs) follows the same pathways of neuronal cell death in
neurodegenerative diseases. Thus, CLIC1 protein increase should be easily detected in
the retina immunocompetent cells system, following a chronic condition of oxidative
stress that is belive to anticipate RGC apoptosis.
1. MECHANISMS OF NEURODEGENERATION
C. Neuro Inflammation
Nome
GABRIELA CONSTANTIN, M.D., Ph.D.
Assistant Professor of Pathology
Contatti
Email: [email protected]
Tel: 39-045 8027102; 8027120; Fax: 39-0458027127
Department of Pathology, University of Verona, Strada le Grazie 8,
37134, Italy
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Role of Inflammation mechanisms in neurodegenerative diseases
In recent years growing evidence show that blood-brain barrier (BBB) breakdown and inflammatory
responses, may initiate and/or contribute to a ‘‘vicious circle’’ of the disease process, resulting in
progressive synaptic and neuronal dysfunction and loss in disorders such as Alzheimer’s disease (AD),
Parkinson’s disease, amyotrophic lateral sclerosis, multiple sclerosis, and others (Zlokovic, Neuron 2008).
In fact, it is clearly emerging that a better understanding of AD inflammatory and immunoregulatory
processes may lead to the development of anti-inflammatory approaches that may help to slow-down or
block the progression of this devastating disorder.
Our group has established expertise in neuroinflammation/neuroimmunology field and will perform an
interdisciplinary study to explore the role of vascular inflammation mechanisms and the role of
leukocytes and leukocyte trafficking in the pathogenesis of AD.
The following main research lines will be developed:
1. Characterization of vascular inflammation and leukocyte-endothelial interactions in animal models of
AD. Our team has previously shown that leukocyte adhesion to brain endothelium leads to BBB damage
and contribute to neuronal loss (Fabene et al., Nat. Med. 2008). We will first study BBB leakage and
vascular inflammation by identifying adhesion mechanisms able to mediate leukocyte-endothelial
interactions in brain vessels. Then we will study the selectins, mucins, integrins and chemokines involved
in rolling and firm arrest of leukocyte subpopulations such as neutrophils, monocytes, Th1 and Th17 cells,
which were able to interact with endothelium in brain microcirculation of AD mice in preliminary studies
performed by our team. To this aim, we will use our established expertise in: in vivo staining, in vivo
Imaging System IVIS® 200 (Caliper Life Sciences) and intravital microscopy in cortical microcirculation
(Constantin et al., Immunity, 2000; Battistini et al., Blood 2003; Pluchino et al., Nature 2005; Fabene et al.,
Nat. Med. 2008; Bolomini-Vittori et al., Nat. Immunol., 2009; Deban et al., Nat. Immunol., in press). Finally,
we will study the effect of adhesion mechanism blockade whether by inhibitory antibodies or genetic
deficiency on the induction and evolution of AD and will identify new potential therapeutic targets for AD.
2. Study of neuro-imuno interactions by using two-photon laser scaning microscopy (TPLSM) in AD
cortex. Leukocyte motility behavior inside the cortex of transgenic animals with AD will be studied in
relation to the vicinity of blood vessels, amyloid deposition and presence of neurofibrillary tangles. Taking
into account our preliminary confocal microscopy studies showing that monocyte, Th1 and Th17 cells
migrate into the cortex of AD mice, we will study the cell-cell contacts between immune cells and neural
cells using transgenic mice expressing fluorescent neurons or glial cell populations. This research line will
be developed based on our TPLSM expertise (Rossi et al., submitted) and our in vivo confocal microscopy
facility, which consists of a customized upright Leica TCS SP5 AOBS system formed by: microscope DM6000
equipped with a complete set of PlanApo objectives, ultrafast scanning head SP5 AOBS coupled to Ar-
HeNe sources and a mode-locked Ti:Sapphire Chameleon Ultra II laser (Coherent Inc).
Overall these studies represent a novel and original contribution to the understanding of AD pathogenesis
and have the potential to identify new therapeutic targets in AD.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Leukocyte recruitment: molecular mechanisms and pathological aspects
Fondazione Cariverona
Durata progetto
February 1, 2009 - January 30, 2011
Abstract del progetto
The research activity includes 3 tasks: Task 1. Mechanisms involved in the
control of granulocyte and monocyte recruitment by Src tyrosin kinases;
Task 2. Role of Rap1, cdc42 and PIP5KC in the control of the modalities of
integrin activation by chemiotactic factors in T and B lymphocytes; Task 3.
Role of Src-family tyrosin kinases in the migration of leukocytes in
inflamed brain vessels and in the induction of neurodegenerative diseases
Adipose-derived stem cells (ADSC) therapy in experimental autoimmune
encephalomyelitis
National Multiple Sclerosis Society, New York, USA
July 1, 2009- June 30, 2011
This project has two objectives. The first objective is focused on the study
of the clinical and neuropathological effect of ADSC on experimental
autoimmune encephalomyelitis (EAE), which represents the animal model
of multiple sclerosis, a neurodegenerative disease. The second objective
is to study the immunomodulatory mechanisms controlling the effect of
ADSC on EAE.
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Name
Contacts
Istituto/Dipartimento
Marina Bentivoglio, MD, Professor
Office: +39-045-8027158; fax: +39-045-8027163; e-mail:
[email protected]
Department of Neurological, Neuropsychological, Morphological and
Motor Sciences – Medical Faculty – University of Verona
Strada Le Grazie 8, 37134 Verona, Italy
Research proposal
Regional vulnerability of the aging brain to neurodegeneration and immune regulatory mechanisms
Background and aims: A wealth of evidence in the last years has indicated that normal aging is hallmarked
by low-grade chronic inflammatory activity, with increased production of proinflammatory cytokines both
peripherally and in the brain, and decreases of anti-inflammatory mediators. Data from our laboratory
(e.g. Deng et al 2006; Sadki et al., Neurobiol. Aging, 28, 296-305, 2007. Xu et al., Brain Behav. Immun. 1,
138-152, 2010) have shown in mice an increase of the immune responsiveness of the aging brain (at the
molecular and cellular levels, the latter including glial cells and T-cell recruitment). This is in contrast with
the known peripheral decline of innate immunity during senescence, and could be involved not only in
cognitive decline but also in the vulnerability of the aging brain to neurodegeneration. On the other hand,
and importantly, not only aging is the primary risk factor for neurodegenerative diseases, including
Alzheimer’s disease (AD), but also these diseases are hallmarked by regional vulnerability in the brain,
which is still elusive. It is, therefore, of primary interest to assess whether immune regional changes in the
aging brain are implicated in its susceptibility to neurodegeneration. To this purpose, the current proposal
aims at investigating: i) regional immune-regulatory changes in the aging brain, and ii) the significance of
such changes in terms of vulnerability of different brain regions to neurodegenerative insults during
senescence.
Plan of work: Step 1: Analyses of the following parameters in mice of young age versus an advanced age,
and in transgenic mice which provide AD models: gene profiling (by means of microarrays and quantitative
real-time RT-PCR) of immune-regulatory genes (including adhesion molecules, complement, cytokine and
chemokine receptors, MHC antigens, Toll receptors, etc.), and immunophenotyping of cells (microglia,
astrocytes, perivascular macrophages, T-cell subtypes, dendritic cells) in different dissected brain regions
(neocortex, hippocampus, hypothalamus, midbrain, etc); magnetic resonance imaging with ultra-small
superparamagnetic particles of iron oxide (USPIO) to visualize regional cellular infiltration. Step 2: A:
Application of challenges with different potentially beneficial or detrimental approaches (proinflammatory
stimuli, like lipopolysaccharide chronic administration; anti-inflammatory therapy; enriched environment,
which is known to exert a beneficial effect; chronic sleep restriction -frequent during normal aging and
known to affect cognitive performance), and test the outcome of these challenges on immune-regulatory
gene expression, neurodegeneration and behavior/cognitive performance. B: Investigate the eventual
occurrence of neurodegeneration during aging in mice with targeted deletion of immune molecules (e.g.
STAT-I signaling pathway knockout, interferon-gamma or its receptor knock-out, etc.) versus wild-type
littermates.
Identified area of interest
Current funding
Project title
Prediction of cognitive properties of new drug candidates for
neurodegenerative diseases in early clinical development (Acronym:
PHARMA-COG). IMI/115009 - IP (Integrated project)
Funding agency
Project duration
European Commission
01/24/2010- 01/23/2014
(note: murine models of AD are available in the framework of this project)
Project abstract
Recently the EU Council of Ministers for Health underlined the
importance of generating novel therapeutic agents both for symptomatic
and disease modifying treatment of Alzheimer’s disease (AD). However,
despite the increase in translational medicine activities attrition rates still
remain high and progress in bringing these biomarkers and models to a
state of readiness as effective decision making tools is slow as each
academic and pharmaceutical company work in isolation.
Bringing together European experts in technologies fully translatable
from animal to human, experts in translational medicine, drug discovery
and mathematical modelling, PHARMA-COG proposes to accelerate this
validation using a ‘MATRIX’ approach i.e. conducting parallel experiments
in animals and human using a comprehensive and standardised battery of
behavioural, neurophysiological, morphological/functional imaging, and
biochemical endpoints to:
o
o
o
o
develop models with greater predictive capacity for the clinics
develop and validate translatable pharmacodynamic markers to
support dose selection
develop challenge models to support early hint of efficacy studies
identify and validate of markers of disease progression and patient
stratification.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Proposta di ricerca
Michela Matteoli
02 50317097
[email protected]
Farmacologia Chemioterapia e Tossicologia medica
Toxic effects of beta-amyloid mediated by microglial cells
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Neurotoxic stimuli
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Elisabetta VEGETO
Via Balzaretti, 9
02 50318263
[email protected]
Dept. Pharmacological Sciences, University of Milan
Gender and hormonal/endocrine signals in neuroinflammation: role in
Alzheimer’s disease and brain aging
Publications from the laboratory demonstrated a key role for estrogens in
regulating microglia cells reactivity in an experimental model of
Alzheimer’s disease and in other neuroinflammatory conditions.
Accordingly, we showed that hormonal manipulations, achieved through
experimental models of menopause and estrogen therapy, impinge on
neuroinflammatory signs in the aging female brain. Considering that
menopause is associated with a higher risk to develop AD in women and
that estrogens are consistently associated with neuroprotective effects,
our research is focused on the identification of the cellular and molecular
targets of estrogens that modulate the susceptibility to
neuroinflammation. Since estrogens action is mediated through the
interaction with specific intracellular receptors, our studies are aimed at
identifying selective estrogenic drugs and appropriate biomarkers of
hormone signaling in neuroinflammation.
Estrogen activity in microglia as a potential pharmacological target for AD
therapy (GP0127Y01)
Telethon; 126,000 Eu
3 yrs (2001-2003)
This study was aimed at evaluating estrogens activity on the activation of
microglia cells that is evoked by beta-amyloid deposition in the brain of
APP23 mice, an animal model of AD. Results show that withdrawal of
circulating estrogens through ovariectomy leads to an increased number
of amyloid deposits that show reactive microgliosis, whereas estrogens
replacement reduce this event to control levels. These results support
the hypothesis that sicrulating estrogens are beneficial signals that acting
also on the neuroinflammatory component of AD delay pathological signs
of this disease.
Estrogens and Women Ageing (Contract no. 518245) Specific Targeted
Research or Innovation Project
European Commission; 510,000 Eu
3 yrs (2005-2007)
This project investigated on the efficacy of estrogenic drugs in reducing
the neuroinflammatory response in the brain of aging female mice, with
particular emphasis on the timing of drug administration in relation with
menopause induction. Results show that menopause potentiates the
neuroinflammatory response in brain and that the anti-inflammatory
activity of several estrogenic compounds is reduced in parallel with
increasing time of hormone deprivation. These results implicate that
hormone replacement therapy might benefit from the anti-inflammatory
activity of estrogenic compounds in brain only when therapy initiates
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
closer in time with the onset of menopause.
MADRI (Menopause: decreased response to increasing inflammation) 5
R01 AG027713-02
NIH; 200,000 Eu
2007-2011
This project investigates more directly on the effect of menopause and
aging on the inflammatory response of the brain. Preliminary results
show that menopause modifies the susceptibility of brain inflammatory
cells that is already altered by the aging process.
DIMI (Diagnostic Molecular Imaging ) A Network of Excellence for
Identification of new imaging markers for diagnostic purpose
European Commission; 376,000 Eu
2006-2010
This project is aimed at challenging imaging techniques for improving
studies on brain reactivity, including inflammation. Several reporter and
transgenic animal models were generated under this program; the main
results from this project show that it is now feasible to trace estrogen
transcriptional activity in brain
Role of estrogens in brain vulnerability to neuroinflammatory signals
associated with aging and neurodegenerative diseases
Ministero della Salute; 20,000 Eu
2009
Aim of the study was to analyse the reactivity of macrophage cells from
male and female mice in response to pathological signals that arise in the
aging brain.
Nome
Dr. Barbara Viviani
Prof. Marina Marinovich
Dr. Mariaserena Boraso
Prof. Corrado L. Galli
Contatti
Tel: +39 (0)2 50318356
Istituto/Dipartimento
Proposta di ricerca
Mail:
[email protected]
[email protected]
[email protected]
[email protected]
Department of Pharmacological Sciences, University of Milan
Role of pro- and anti-inflammatory cytokines in the prognosis
and progression of neurodegenerative diseases
Area di interesse
identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
MOLECULAR MECHANISMS OF INFLAMMATION AND REPAIR IN STROKE:
NEURORADIOLOGY AND BIOCHEMICAL ENDPOINTS TO DESIGN A NOVEL
THERAPEUTIC AND PROGNOSTIC APPROACH
Regione Lombardia
Durata progetto
24 mesi
Abstract del progetto
Stroke remains a leading cause of adult death and disability, which still demand an
appropriate therapeutical approach. Some degree of spontaneous behavioral recovery is
usually seen in the weeks after stroke onset. In general, the best outcomes are associated
with the greatest return toward the normal state of brain functional organization.
Reorganization of surviving central nervous system elements supports behavioral
recovery. In this framework, the development of therapeutic approaches whose goal is to
promote repair and restoration of function after stroke could be strategic. Two of the
most prominent molecular events occurring in brain ischemia are inflammation and
overactivation of the NMDA subtype of glutamate receptors (excitotoxicity), which lead to
the progression of neuronal damage. Although this, an endogenous neuroprotective
response, at least partially mediated via induction of neuroprotective factors, is also
elicited by ischemic injury. Actually, the great potential of the brain for reorganization,
plasticity and repair after injury is a critical issue to be taken into consideration, that has
previously been understimated.
Knowing the pathophysiological mechanisms that follow ischemia and their time course is
essential in order to be able to identify and use biomarkers in monitoring the patients
with acute stroke. Although still in the research phase, some biomarkers have been
shown to correlate with the clinical evolution and could be important in foreseeing the
clinical response to the treatment of the acute phase and to the different rehabilitation
strategies in the post-acute and chronic phases. In addition, the inflammatory and
neuroprotective response during the evolution of stroke can be monitored in the patient
through suitable techniques such as neuroimaging.
Up to now research on molecular targets in cerebral ischemia for developing novel
therapeutics has mainly focused on single pathways, being inflammation or excitotoxicity
or oxidative stress. The step forward, to go beyond the current state of the art, of the
present project is to try to consider a group of different concurrent factors elicited during
stroke. The project aims at investigating the cross-talk between inflammation,
excitotoxicity, and endogenous neuroprotective/neuroregenerative pathways. In
collaboration with the Neurological Institute C. Mondino, levels of selected biomarkers
representative of inflammation and neuroprotection will be monitored in the serum of
patients with stroke at different time points and then will be correlated with
neuroimaging data and the clinical outcome. This will
provide a clinical basis to finalize the basic research focussed on the
evaluation of the molecular mechanisms recruited in the cross-talk
between inflammation, excitotoxicity, and endogenous
neuroprotective/neuroregenerative pathways. To this purpose we will
evaluate in glial and neuronal population (i) the impact of inflammation
on expression of neuroprotective factors relevant to brain ischemia (i)
the effect of neuroprotective factors on inflammation, (ii) the effect of
cytokines and neuroprotective factors on excitotoxicity driven by the
NMDAR
The project is designed to identfy new prognostic markers and new
targets for therapy, aimed at boosting or superinducing the endogenous
neuroprotective response directly or interfering with the mechanisms
that blunt this response in conditions of injury.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
ANGELINI - ACRAF SpA
Claudio Milanese
06-91045290
[email protected]
Angelini Research Center- S.Palomba (Rome), Italy
Study of the role of pro-inflammatory chemokines in
neuroinflammatory disorders and analysis of the therapeutic potential
of chemokine synthesis inhibitors
Abstract
Chemokines and their receptors play a crucial role in the trafficking
of leukocytes and other immune system cells and are of particular
interest in the context of the unique immune responses elicited in
the central nervous system (CNS). Within this particular district,
most data focus on the role of CCL2 and the related factors CCL7
and CCL8, since they have been implicated in a wide range of
neuropathologies, including trauma, ischemic injury and multiple
sclerosis (Semple BD et al., 2010, J Cerebral Flow & Metabolism
30:459). Furthermore, members of this chemokine family and their
receptors recently emerged as key modulators in nociceptive influx
transmission in neuropathic and inflammatory chronic pain models
(Gosselin RD et al., 2008, Curr Med Chem 15:2866), providing
evidence of the involvement of chemokines as important mediators
of the initiation and maintenance of pain hypersensitivity.
Angelini has a large experience in the study of chemokine synthesis
inhibitors.
A small original Angelini molecule able to inhibit the synthesis of
CCL2, bindarit, has recently successfully concluded a phase II clinical
study in diabetic nephropathy and it is currently undergoing
another phase II study in restenosis. The aim of the present project
is the identification and development of new molecules able to
inhibit the synthesis of chemokines and endowed with physicochemical and ADME characteristics useful to target CNS disorders.
The evaluation of the pharmacological activity of such compounds
in appropriate models will allow to assess their potential as
therapeutic agents useful for the treatment of CNS disorders and
chronic pain.
ISTITUTO SUPERIORE DI SANITA’ (Referente, Luisa Minghetti)
The Istituto Superiore di Sanità is the leading technical and scientific body of the Italian National Health
Service. Its activities include research, clinical trials, control and training in public health. The Institute
conducts scientific research in a wide variety of fields and also serves as a major source of information
relating to public health and biomedicine in Italy through online connections to national and international
scientific databases and data banks.
The institute has been actively involved in research in ageing and related neurodegenerative diseases over
the last decade. It is partner in the European Research Area in Ageing network (ERA-AGE and ERA-AGE2)
funded under the Sixth and the Seventh Framework Programme.
The research on Alzheimer’s disease in ISS takes advantage of multidisciplinary expertise and resources
including proteomics, molecular and cellular biology, neuroimmunology, behavioral analysis, in vitro and in
vivo models, genetic, clinical and epidemiological studies, advanced statistical analysis, data and bio-banks.
Main topics of research are: in vitro toxicity of amyloid proteins; excitotoxicity and free radical-related
alterations of signaling pathways and post-translational modifications of post-synaptic machinery; analysis
of protein aggregate characteristics responsible for the induction of amyloid formation; characterization of
relevant rodent models by pre-clinical imaging with MRI scanning; characterization of behavioral and
cognitive alterations in relation to neurochemical/neuroanatomical markers in AD murine models; effects
of diet, pharmacological agents, or neurotoxicants on disease susceptibility and progression in AD murine
models; early environmental factors in modulating onset and progression of neurodegenerative diseases;
self antigens and autoantibodies in the pathogenesis of AD and as biomarkers of the disease; peripheral
oxidative stress and antioxidant defense related biomarkers as prognostic and diagnostic tools;
identification of mutations and polymorphisms affecting clinical phenotype and progression; descriptive,
analytical, clinical epidemiological studies; longitudinal data bases for cohorts of elderly persons with
cognitive deficits (IPREA and ILSA).
Research projects currently active include:
• Role of Genetic Determinants on the Progression of Clinical Phenotype from MCI to Alzheimer’s Disease.
PI: A.Confaloni (National coordinator F.Tagliavini), Italian Ministry of Health .Eur 60,000
• Determinanti genetici e fattori modulatori nelle malattie neurodegenerative: modelli clinici ed animali P.I. A. Confaloni (National coordinator A. Bruni), Italian Ministry of Health, Eur 70,000
• Risk Assessment of iatrogenic transmission of neuro-AMYloidoses (RAAMY). PI: M.Pocchiari (in
collaboration with E.Comoy, CEA, France), Alliance BioSecure, Fondation de recherche reconnue d’utilité
publique.
• Early diagnosis of Alzheimer’s diseases through the combination and correlation of molecular, functional
and clinical biomarkers PI: M.Pocchiari, Italian Ministry of Health. Eur. 364,000
• Innovative approaches to the study of induction/repair of DNA oxidative damage in models of
neurodegenerative diseases: molecular basis and identification of possible therapeutic targets. PI:
P.Popoli, Italian Ministry of Health. Eur 364,000
• Effects Of Dog-Assisted Therapies On Physical And Psychological Well Being In The Institutionalized
Elderly. PI: F.Cirulli, ISS and Nando Peretti Foundation, Eur 100,000
• Therapeutic effects of human-animal interactions in a rural environment on neuropsychiatric diseases
and brain aging. PI: F.Cirulli, INEA Eur 62,000
• Nuovi farmaci ad azione neurotrofica e neuroprotettiva per applicazioni terapeutiche nelle neuroscienze:
malattia di Azheimer, malattia di Huntington e Sclerosi Laterale Amiotrofica. PI: D. Merlo, (National
coordinator A. Cattaneo). MIUR – FIRB “Idee Progettuali RBIP063ANC” 2007, Eur 32,000
• Ruolo neuroprotettivo della Timosina beta4 in modelli sperimentali in vitro di eccitotossicità, ischemia e
Alzheimer Disease”. PI: D. Merlo, Tecnogen Spa, Eur 125,000
• Role of recurrent herpetic infections in neuronal damage. PI D. Merlo (National coordinator
A.T.Palamara), Ministero della Salute, Eur 70,000
• An integrated approach to identify functional, biochemical and genetic markers for diagnostic and
prognostic purposes in the elderly, in the centenarians and in people with dementia, Alzheimer’s disease,
mild cognitive impairment. PI: N.Vanacore, Italian Ministry of Health, Eur 291,000
• Alzheimer's Disease (AD) and antipsychotics: a long term, multicentre, double blind, randomised clinical
trial. PI: R.Raschetti, Italian Agency of Drug, Eur 1,950,000.
• EC 7th Framework Programme “ERA AGE 2 Project – European Reasearch Area in Ageing Extension” .
E. Scafato.
EC 7th Framework Programme “ FUTURAGE Project – A Roadmap for Ageing Research” . E. Scafato
• EC 2nd Programme of Community Action in the Field of Health “VINTAGE Project – Good Health into
Older Age”. E. Scafato.
• EC 5th Framework Programme “DESCRIPA Project - Development of screening guidelines and criteria for
predementia Alzheimer’s disease”. E. Scafato.
Nome
Paola Bossù
Contatti
Via Ardeatina 306, I-00179, Rome
Tel.
06-51501520
E mail
[email protected]
Istituto/Dipartimento
IRCCS Fondazione Santa Lucia/Clinical and Behavioral Neurology
Proposta di ricerca
Role of inflammation in Alzheimer’s Disease: study of pro-inflammatory cytokines for the
identification of new pathogenic and diagnostic targets
Area di interesse
Inflammation and neurodegeneration
identificata
Finanziamenti ricevuti
Titolo progetto
Genetic Risk factors and peripheral biological markers of conversion
from Mild Cognitive Impairment to Alzheimer’s Disease
Ente finanziatore
Italian Ministry of Health
Durata progetto
01/2009 - 01/2011
Abstract del progetto
Several studies have demonstrated that inflammation plays an
important role in mediating glial alterations and neurodegeneration
that occur in AD patients. This phenomenon includes the accumulation
of reactive microglia and astrocytes in damaged regions of AD brain
and increased pro-inflammatory cytokine expression that may further
contribute to the development and progression of pathological state.
Accordingly, in several studies pro-inflammatory cytokines such as
Interleukin (IL)-1, IL-6, Tumor Necrosis Factor (TNF)-α and IL-18
have been found to be peripherally increased in AD patients and
substantially impaired in the late stage of the disease. However, the
temporal definition of their implication in respect to disease
progression, as well the extent to which inflammatory factors
participate to the neuronal damage are issues of fundamental
importance, which are to date still unclear. Recent studies performed
by this participant unit have identified a possible role for the proinflammatory cytokine IL-18 in AD, by means of genetic data
suggesting that IL-18 gene promoter polymorphisms can predict risk
and outcome of AD, and by demonstrating that IL-18 is overexpressed
by blood cells of AD patients and correlates with their cognitive
decline. In the present study, by using a longitudinal clinical approach,
the previous results regarding IL-18 will be expanded also to other
inflammatory mediators with potential role in the disease progression
and pathogenesis. In fact, this project is aimed to investigate the
impact of pro-inflammatory and anti-inflammatory parameters during
disease progression in patients at the higher risk of developing AD, i.e.
MCI patients. These objectives are relevant for the identification of
new diagnostic markers of the AD preclinical form and for the
comprehension of inflammatory mechanisms, likely participating in
the earliest pathogenic processes of AD.
1. MECHANISMS OF NEURODEGENERATION
D.
Oxidative Stress and neurodegeneration
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Elisa Greggio, Marco Bisaglia, Luigi Bubacco
Via Ugo Bassi 58/b, 35129 Padova, ITALY
Tel. +39 049 8276244
Dipartimento di Biologia, Università di Padova
Indagine dei meccanismi molecolari alla base della malattia di Parkinson
Mitochondrial dysfunction and axonal transport in genetic models of
Parkinson’s disease
MIUR
4 years
Mitochondrial dysfunction and oxidative stress have been implicated
in the pathogenesis of Parkinson’s disease (PD). Defective axonal
transport, including abnormal accumulation of misfolded proteins and
organelles and damage of transport of cargoes through axons, is also
observed in many neurodegenerative diseases. Few lines of evidences
suggest that both alpha-synuclein and LRRK2, proteins involved in
dominantly transmitted forms of familial PD, may play a role in
mitochondrial function, microtubulo stability/dynamics and axonal
transport. The aim of this project is to use cellular and, in part, animal
models of !-synuclein and LRRK2 to explore the
physiological and pathological role of this two genes in mitochondrial
functionality and in the transport of mitochondria and synaptic vesicle
along the axons.
Biochemical characterization of full length human recombinant LRRK1
and LRRK2
Michael J Fox Foundation
1+1 years
Mutations in one gene, LRRK2, are a common cause of Parkinson’s
disease. However, the protein product of this gene is large and complex
and, to date, no one has been able to make highly pure, functional LRRK2
that can be easily used for biochemical experiments. The primary
objective of this project is to make purified protein by combining
expertise in multiple technologies from each of our collaborating
laboratories and to share our methods with other researchers. If
successful, our secondary objective is then to perform an exhaustive
search for other proteins that may interact with LRRK2.
Molecular mechanisms that involve the oxidation product of dopamine
in the pathogenesis of Parkinson disease.
Prin- MIUR
1+1 years
Several hypotheses have been proposed in the literature to describe
the mechanisms that lead to neurodegeneration in PD. This project will
focus on the ossidative stress associated to the redox chemistry of
dopamine. The rational in this choice is that dopamine oxidation may
reasonably account for the specificity observed for dopaminergic neuron
degeneration in PD. Although the oxidation of DA and DOPA lead to the
formation of toxic ROS, the main cytotoxic species which could potentially
explain the specific damage to dopaminergic neurons remain the highly
reactive quinone species (DAQ) that have many potential protein targets
for chemical modifications. The identification of several proteins, whose
functions are affected by the mutations associated to familiar PD,
triggered the hypothesis that late onset idiopathic forms of PD may
derive from similar functional effects induced by chemical modifications
produced by DAQ. This project will be focus on the definition of the
damages induced by DAQ on two cytoplasmatic (DJ-1 and α-Synuclein)
and two mitochondrial (PINKI and Superoxide Dismutase 2) possible
protein targets.
Nome
Contatti
Antonella Bobba
Tel: 080 5442412 Fax: 0805443317 E-mail: [email protected]
Istituto/Dipartimento
Istituto di Biomembrane e Bioenergetica (IBBE), CNR
Via Amendola 165/A 70126 Bari, Italy
Proposta di ricerca
Area di interesse
identificata
MITOCHONDRIA,
APOPTOSIS AND NEURODEGENERATION:
MITOCHONDRIAL DYSFUNCTION IN NEURODEGENERATIVE DISEASES
CHARACTERIZATION
OF
There is compelling evidence for the direct involvement of mitochondria in
neurodegenerative disorders. In most cases, increased oxidative stress appears to
be the crucial initiating event that affects respiratory chain function and starts a
vicious cycle finally leading to neuronal cell death via apoptosis or necrosis. In
addition a functional interaction exists in the cells between cytoplasm and
mitochondria and the cross-talk ability of these cellular components is involved in
cell homeostasis and death. In the light of this, the goal of this topic is to ascertain
whether and how, during neurodegeneration, the energy metabolism in the
cytoplasm changes as a result of mitochondrial alterations and vice-versa and the
mechanism by which certain compounds can prevent cell death.
CHARACTERIZATION OF THE MOLECULAR MECHANISMS UNDERLYING AGING, CELL SENESCENCE
AND NEURODEGENERATIVE DISEASES
Aberrant modulation of cell signaling pathways involved in cell death regulation
underlies several human pathological conditions, such as neurodegeneration.
Although the mechanistic aspects of cell death and growth have been extensively
investigated, there is still lack of knowledge about the early events that lead to
deregulation of cell homeostasis and about the mechanisms of cell sensitization to
growth and death signals in response to changes in cell environment. Thus, it is
worthwhile to characterize the progression of the biochemical events and in
particular of those occurring in the early phase of neurodegeneration with the aim
to identify the critical steps that once blocked could allow for the restore, even if
partial, of the impaired cellular function/activity. The characterization of the main
signalling pathways that are activated and/or impaired during neurodegeneration
and the close definition of the cause-effect relationship between them are powerful
tools which are potentially usable to design new diagnostic, prognostic and
therapeutic approaches. To this aim it is of great relevance an approach in which
functional genomics and whole-genome expression profiling are combined in order
to efficiently identify key players in biological response pathways. This is possible if
different cellular systems are complementarily used in the experimental approach.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Neurotrofine e meccanismi relativi a malattie neurodegenerative.
UO3 - Stress ossidativo e bioenergetica mitocondriale nella patogenesi delle
malattie neurodegenerative
MIUR (Progetto CNR/MIUR - Fondo FISR)
2 years (11/06/2003 – 1/10/2005)
Studies on the involvement of mitochondria and role of oxidative stress in the
ethiopathogenesis of neurodegenerative diseases by using primary neuronal cell
cultures as in vitro model. The main results can be summarized as follow: i) early
impairment of the energetic metabolism; ii) released cytochrome c exerts a crucial
role by acting both as a ROS scavenger and an electron donor other than a caspase
activator; iii) ATP level increases in the early phase of the death process and iv) ROS
and thereafter activated caspases induce the progressive impairment of the
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
mitochondrial ANT.
Malattie neurodegenerative come conseguenza di un alterato processamento di
proteine neuronali, modelli animali e di colture cellulari in vitro
MIUR (Progetto FIRB cod. RBNE01ZK8F)
3 years (08/01/2003 – 10/01/2007)
Both animal and in vitro cultured cellular models have been used to study the
molecular mechanisms which are at the basis of some neurodegenerative diseases
and to identify new, potential therapeutical tools. Studies have been mainly focused
on a thorough analysis of the mitochondrial function impairment with collapse of
oxidative phosphorylation and increased production of reactive oxygen species.
Bioenergetic and apoptotic systems of mitochondria. Genomics, proteomics, cellular
homeostasis and physiopathology.
WP2- Mitochondria role in animal, plant and microorganism cell apoptosis
MIUR (Progetto MIUR N. 157)
3 years (2006-2009)
The aims of the research project are concerning with: a)The role of mitochondria in
the progression of neuronal apoptosis; b)The role of mitochondria in the apoptosis
of a plant model system; c)The role of mitochondria in yeast apoptosis; d)The role of
mitochondria in the induction/resistance to apoptosis in neoplastic cells
Molecular bases in ageing-related degenerative syndromes
MIUR - Progetto MERIT - Protocollo: RBNE08HWLZ
(UR n 11: Protocollo: RBNE08HWLZ_012)
3 years (approved but not yet funded)
The aim of this project is to discover specific molecular pathways and/or targets for
the developing of new therapeutic strategies. Research activities are grouped into
four workpackages: 1) Aggregation states and cytotoxic activity: molecular and in
vivo/in vitro biological studies. 2) Oxidative stress and ageing: biochemical
processes and regulation of gene expression. 3) Dismetabolic factors and
neurodegeneration: molecular, biophysical and biochemical studies. 4)Exogenous
and endogenous molecules against ageing and neurodegeneration: application for
the diagnosis and therapy.
Biotechnology applied to the study and utilization of Apulian extra-virgin olive oil
and its minor components in aging and in diseases associated to oxidative stress
Regione Puglia (Progetto Reti di Laboratorio Pubblici di Ricerca)
3 years (approved but not yet funded)
Constitution of a Laboratories Network aimed to identify the nature and efficacy of
minor components of Apulian extra virgin olive oil in the prevention of oxidative
stress-associated pathologies
Nome
Lenaz Giorgio and Giancarlo Solaini
Contatti
051-2091229; 333-2383218; fax: 051-2091217
[email protected]
051-2091215 ; 339 1948613 ; fax 051-2091224
[email protected]
Istituto/Dipartimento
Dipartimento di Biochimica
Proposta di ricerca
The involvement of mitochondria in neurodegenerative diseases, including Alzheimer disease (AD) is
widely documented, although no clear causal relationships are available for a great number of
scattered observations. Both a decrease of mitochondrial respiration, in particular of cytochrome
oxidase and an enhanced production of Reactive Oxygen Species (ROS) has been reported in autoptic
samples of AD patients and in neuronal cells of animal models of AD. Most researchers in the field
believe these findings being the consequence of toxic amyloid peptides (A-beta) interacting with
mitochondrial structures, and several studies have recently shown that A-beta and tau proteins,
characteristic of AD, interact with mitochondrial respiratory complexes, opening novel hints to
understand the pathogenesis of the disease. A significant investigation on the above issue has been
carried out by some of us, but it’s noteworthy that our research group has a world wide reputation
on the study of mitochondrial structure and function and has already been involved in research on
aging and neurodegenerative diseases including AD. In particular we have shown the functional
consequences of the recently discovered super-association of respiratory complexes and
demonstrated that these super-complexes are easily dissociated by a number of changes in proteinprotein and lipid-protein interactions. We intend to propose and test the following working
hypothesis. Under conditions likely present in brain regions of AD patients, the interaction of A-beta
and tau with mitochondria at the level of respiratory chain is able to disrupt super-complex
associations with consequent destabilization of the individual complexes, in particular Complex I
(NADH Coenzyme Q reductase). This would necessarily determine an enhancement of production of
ROS, thus explaining the observed oxidative stress in AD mitochondria. Loss of efficient electron
transfer and altered assembly of Complex I would determine decreased oxidative phosphorylation
and at the same time the oxidative stress could represent a sufficient stimulus for triggering cell
death by necrosis or apoptosis. Our laboratory has the expertise and technology for all investigations
required to test this hypothesis, and we are currently setting conditions considered likely to occur in
brain regions of AD patients. The study will be performed on animal model systems and in human
cells cultures. On the other hand, a better understanding of AD pathogenesis might establish the
basis for attempts of therapeutic interventions, including metabolic therapy as recently shown in our
laboratory for diseases featuring alterations of ATP synthesis.
Area di interesse identificata
Finanziamenti ricevuti
Basic research/new treatment strategies
Titolo progetto
Ruolo dell’organizzazione sopramolecolare della fosforilazione
ossidativa mitocondriale nella produzione di specie reattive
dell’ossigeno e in alterazioni patologiche in sistemi modello e in
cellule umane.
Ente finanziatore
MIUR (PRIN 2008)
Durata progetto
2 years
Abstract del progetto
In this project we will test the working hypothesis that an initial
enhanced ROS generation due to different possible reasons and
originating in different districts of the cell besides mitochondria
would induce disorganization of the supercomplex assemblies of the
oxidative phosphorylation system, eventually leading to instability
of Complex I quaternary structure; both the lack of efficient electron
channelling and the decrease of Complex I would cause a fall of
NAD-linked respiration and ATP synthesis, leading to pathological
changes. This hypothesis will then be tested in two pathologies in
which a major pathogenic factor is oxidative stress, i.e. the NARP
syndrome (neuropathy with ataxia and retinitis pigmentosa) due to
mitochondrial DNA mutations in the ATP6 gene, and diabetic
peripheral neuropathy, a common and severe complication of
diabetes mellitus. (Details follow)
Nome
Contatti
Annalisa Santucci
Tel. +390577234958
Fax +390577234903
Email [email protected]
Department of Molecular Biology, University of Siena, ITALY
Istituto/Dipartimento
Proposta di ricerca
Amyloid β peptide (Aβ) is directly involved in the pathogenesis of Alzheimer Disease (AD) or tightly
correlated with other primary pathogenic factors. The predominant forms of Aβ in the human brain are
Aβ(1-40) and Aβ(1-42), but Aβ(25-35) fragment, physiologically present in elderly people [1], is the more
toxic region and has been recently found to play a relevant role in AD, due to its peculiar aggregation
properties. In fact, actually, the ability to affect cognitive processes is typical not only of the full-length
peptide, but also of several Aβ fragments, in particular just the undecapeptide Aβ(25-35). This peptide is
regarded as the functional domain of Aβ, responsible for its neurotoxic properties and represents the
actual biologically active region of Aβ. In vivo, Aβ(25-35) is present in neurons of subiculum and entorhinal
cortex of AD brains, and it was also observed in Inclusion-Body Myositis (IBM) muscle. Self-assembly of
Aβ(25-35) is a key player in AD, since it could nucleate the assembly of Aβ monomers and a bent in this
region could be the rate-limiting step in Aβ fibril formation. In virtue of this, there is increased interest in
the role of Aβ(25-35) fragment in free radical-associated neurotoxicity in AD brain. Age-dependent
racemisation of Ser residue at the 26th position in Aβ(1-40) is critical in AD pathogenesis. In vivo
conversion of non-toxic [DSer26] Aβ(1-40) to toxic [D-Ser26]Aβ(25-35) has been finally demonstrated and
the presence of truncated and toxic [DSer26] Aβ(25-35) in AD brains, but not in age-matched control
brains, adds a major value to in vivo neurodegeneration mechanism. Given the suggested centrality of
Aβ(25-35) to the pathogenesis of AD, this peptide should deserve major attention also as a therapeutic
target. In fact, diagnosis and monitoring of sporadic AD have long depended on clinical examination of
individuals with end-stage disease. However, upcoming anti-AD therapies are optimally initiated when
individuals show very mild signs of neurodegeneration. Our research group provided for the first time an
exhaustive overview about Aβ(25-35) [2] and during the last few years it has deepened the aggregation
properties of this peptide according to solution and environmental conditions [3]. Actually, we are
dedicating attention to this peculiar peptide in the light of its great oxidative stress generation capacity
and extreme toxicity in neuronal cells and synaptosomes. The evidence points toward Aβ(25-35) being
primarily responsible for neurodegeneration observed in AD. The question remains on how this can induce
such degenerative effects. Strong of our experience in the field of the study of oxidative stress [4-9], we
propose ourselves to estimate the mechanisms been involved in the ROS generation Aβ(25-35)-mediated
in different cell models. We can use SELDI technology to study [DSer26] Aβ(25-35) as a core biomarker for
the mild cognitive impairment stage of AD. From a longer perspective, establishment of the most effective
combinations of different biomarkers and other diagnostic modalities may be foreseen. Moreover, due to
our well established experience in Proteomics [10-15], we will apply proteomics techniques to study cellspecific early markers of brain aging-related degeneration in human astrocytes and neurons that may
contribute to neurodegenerative damage. The protein expression patterns of the AD neuron and astrocyte
cultures will be compared with those obtained from healthy people. Differentially expressed spots will be
identified by matrix-assisted laser desorption/ionization-time of flight peptide map fingerprinting and
database search. A number of the protein alterations have been previously reported in the brain tissue
proteome of animal models, aged brain or AD brain, but are still quite unexplored in neuron and astrocyte
proteome and our project could help to clarify if both cell types are involved in the brain degenerative
changes as well as to study neuronal changes having a greater influence in the pathology progress. These
findings will be discussed in reference to the effect of specific protein oxidation and changes of expression
on potential mechanisms of abnormal alterations in metabolism and neurochemicals, as well as to the
learning and memory deficits in AD.
References
[1] Kubo, T.; et al. A possible mechanism of neurodegeneration in AD - Involvement of racemized βamyloid. Soc. Neurosci., 1999, Abstr. 25, p. 838.
[2] Millucci L, et al. Conformations and biological activities of Amyloid Beta Peptide 25-35. Curr Protein
Pept Sci. 2009 Sep 15.
[3] Millucci, L., et al. Rapid aggregation and assembly in aqueous solution of Aβ(25-35) peptide. J. Biosci.
2009, 34(2), 293-303.
[4] Braconi D, et al. Proteomics and redox-proteomics of the effects of herbicides on a wild-type wine
Saccharomyces cerevisiae strain. J Proteome Res. 2009;8(1):256-67.
[5] Braconi D, Possenti S, Laschi M, Geminiani M, Lusini P, Bernardini G, Santucci A. Oxidative damage
mediated by herbicides on yeast cells. J Agric Food Chem. 2008 May 28;56(10):3836-45.
[6] Braconi D, et al. Comparative analysis of the effects of locally used herbicides and their active
ingredients on a wild-type wine Saccharomyces cerevisiae strain. J Agric Food Chem. 2006 19;54(8):316372
[7] Desideri G, et al. Effects of bezafibrate and simvastatin on endothelial activation and lipid peroxidation
in hypercholesterolemia: evidence of different vascular protection by different lipid-lowering treatments. J
Clin Endocrinol Metab. 2003;88(11):5341-7.
[8] Desideri G, et al. Angiotensin II inhibits endothelial cell motility through an AT1-dependent oxidantsensitive decrement of nitric oxide availability. Arterioscler Thromb Vasc Biol. 2003;23(7):1218-23.
[9] Trabalzini L, et al. Proteomic response to physiological fermentation stresses in a wild-type wine strain
of Saccharomyces cerevisiae. Biochem J. 2003;370(Pt 1):35-46.
[10] Bernardini G, et al. Postgenomics of Neisseria meningitidis for vaccines development. Expert Rev
Proteomics. 2007;4:667-77.
[11] Bernardini G, et al. The analysis of Neisseria meningitidis proteomes: Reference maps and their
applications. Proteomics. 2007;7(16):2933-46.
[12] Spreafico A, et al. A proteomic study on human osteoblastic cells proliferation and differentiation.
Proteomics. 2006;6(12):3520-32.
[13] Mini R, et al. Comparative proteomics and immunoproteomics of Helicobacter pylori related to
different gastric pathologies. J Chromatogr B Analyt Technol Biomed Life Sci. 2006;833(1):63-79.
[14] Bernardini G, et al. Proteome analysis of Neisseria meningitidis serogroup A. Proteomics.
2004;4(10):2893-926.
[15] Magnani A, et al. Two-step elution of human serum proteins from different glass-modified bioactive
surfaces: a comparative proteomic analysis of adsorption patterns. Electrophoresis. 2004;25(14):2413-24.
Area di interesse identificata
Post-Genomic Biochemistry of Neurodegenerative Diseases
Finanziamenti ricevuti
Titolo progetto
Characterization of peptide oligomers in Alzheimer Disease
Ente finanziatore
Siena Biotech s.p.a.
Durata progetto
1/07/2006 - 30-04-2009
Abstract del progetto
Aβ(25-35) is the shortest peptide sequence that retains biological activity
of full-length Aβ(1-42) and exhibits large β-sheet aggregated structures.
Because of these features, it has often been chosen as a model for fulllength Aβ in structural and functional studies but kinetic and structural
studies of this peptide are indeed strongly hampered by its tendency to
quickly assemble into insoluble aggregates. The aim of the project was to
study the uncommon rapidity with which Aβ(25-35) self-assembles at
different pH. The obtained results support the hypothesis that the early
Aβ(25-35) aggregated forms, that develop at pH 3, may not enhance the
growth of dangerous fibrils generated at pH 7.4. We conclude that the
mechanism by which Aβ forms toxic fibrils is initiated in vivo and probably
does not involve low pH compartments, confirming the hypothesis of Kubo
et al. (2002) on the origin and toxicity of Aβ(25-35).
1. MECHANISMS OF NEURODEGENERATION
E. Trophic factors involvement in the
neurodegeneration process
1. MECHANISMS OF NEURODEGENERATION
F. Endocrine and Metabolic Factors
Country: Italy
Contact person: Paola Fragapane, Cecilia Mannironi (CNR)
Date: 28/01/2010
I) Strategic Issues
Stress is a term to describe experiences that are challenging both emotionally and physiologically.
A feature of the stress response is the activation of the autonomic nervous system and
hypotalamus-pituitary-adrenal (HPA) axis with the production of glucocorticoids hormones. This
response is needed to survive dangerous situation but excessive adrenocortical and autonomic
function is deleterious for health and survival. The central nervous system in response to such
activation elaborates the appropriate behavioral and physiological responses. As regards to the
neuroanatomical substrate the hypothalamus and the brain stem are essential in the response to
stressors, however, other brain regions are also well known targets of stress hormones. Among
such regions, research has focused its attention on the prefrontal cortex, the hippocampus and the
amygdala. Interestingly these brain structures have been related also to functions such as
memory, and decision making but also to anxiety and other psychiatric diseases.
Acute (single stress) and chronic (repeated) stress induce prominent changes in neuronal activity
and synaptic functions. These changes are thought to be due to neuronal remodelling such as
dendrites shortening and debranching (Mc Ewen, 2007) in the hippocampus and in the medial
prefrontal cortex. These stress-induced changes are mediated by modification of gene expression
at transcriptional levels. However, recent studies are pointing out the contribution of
posttranscriptional regulation to stress associated responses (De Rijk et al., 2003). An important
post-transcriptional mechanism of gene regulation involves microRNA (miR). A range of evidence
suggests that these small RNAs form complex networks involved in the regulation of differentiation
and development, via regulation of chromatin modification, transcription, translation and RNA
stability. They are small RNA molecules (about 22 nucleotide long) that inhibit mRNA translation or
induce mRNA degradation by sequence specific pairing with mRNA 3' untranslated regions
(UTRs). In the vertebrate nervous system miR have been shown to play a role during development
and neurogenesis (Kosik, 2009; Lagos-Quintana et al., 2001). MiR are also abundantly expressed
in the adult brain and appear to regulate the maintenance of mature trait and synaptic plasticity
(Mehler and Mattick, 2006). MiR134 was reported to be a regulator of dendritic spine development
in brain (Schratt et al., 2006). Perturbation in miR are associated with a number of neuronal
diseases as X-linked mental retardation, Parkinson’s disease and schizophrenia.
II) Priority areas
The molecular mechanism underlying brain response to stress are still incompletely understood.
The aim of this project is to identify region-specific stress-induced changes in miR expression. The
expression profiles of miRs in brain regions known to be involved in stress response such as the
hippocampus, the amygdala and the prefrontal cortex will be analyzed after the exposure to
aversive stimuli. The analysis will be performed in rodents exposed to single or repeated restraint
stress and gene expression in each brain regions will be compared with that of matched control
mice. LNA (locked nucleic acid) microarrays will generate a limited set of candidate miRNAs, which
will be validated by Northern blots and qPCR. In order to have a better neuroanatomical definition
of miR expression selected miR will be analyzed by in situ hybridization. mRNA targets it will be
predicted through the use of freely available algorithms and will be validated by reporter construct
expression in cell system. Moreover bioinformatic analysis of miR upstream promoter sequences
will allow us to identify transcription factors involved in their modulation in response to stress
stimuli.
III) Impact
The hippocampus, the amygdala and the prefrontal cortex are interconnected and influence each
others via direct and indirect neuronal activity. It is well established their involvement in the stress
response and in the plastic changes occurring in response to acute and repeated stress exposure.
Our analysis of miR expression pattern will allow us to understand possible region specific
expression pattern and/or common molecular networks.
Translation of the information on stress effects from animal models to the human organism is an
important challenge. Indeed maladaptive response to stress is considered causal to many
psychiatric diseases such as major depression, anxiety as well as schizophrenia. The
understanding of the contribution of non coding RNAs might allow on the one hand a better
understanding of the neural mechanisms at the basis of stress response on the other hand might
help identify possible targets for the development of new future therapies.
Nome
Roberto Rimondini-Giorgini
Contatti
[email protected]
Istituto/Dipartimento
Department of Pharmacology
Via Irnerio 48
40126 Bologna
Italy
Proposta di ricerca
Area di interesse identificata
Developing competitive animal models / basic research
Finanziamenti ricevuti
Titolo progetto
MECHANISMS BEHIND THE ASSOCIATION OF APOE4 GENOTYPE
AND LIFE STYLE RELATED FACTORS IN ALZHEIMER`S DISEASE
Ente finanziatore
Fondazione Cassa di Risparmio di Bologna
Durata progetto
2 anni
Abstract del progetto
Alzheimer`s disease (AD) is a devastating neurodegenerative disease
and the most common cause of dementia. The current estimate
number of people with dementia in the world stands at 24.3 million,
with an estimated 4.6 million new cases every year in the world.
There are no definitive diagnostic tests, biological markers of AD or
pharmaceutical treatments. At central nervous system (CNS) level,
the pathological signs include deposits of extracellular amyloid-β
peptide (Aβ) in plaques. Loss of neurons and synapses is also
widespread. Whilst some cases have a genetic component (familiar
AD) the majority of cases (90-95%) have unknown cause and are
named sporadic AD. Familiar AD seems to be correlated to
mutations in key genes such as in the amyloid-precursor-protein
(APP) gene and in the presenilin genes (1 and 2). On the contrary,
sporadic AD seems to be consequence of a complex interaction
between both genetic and environmental risk factors. The major
genetic risk factor is an allelic variant of apolipoprotein E (apoE)
called apoE4. Nowadays, no specific environmental risk factor has
been definitively identified as being associated with AD. Anyway, AD
is associated with a history of depressive illness, traumatic head
injury, cardiovascular disease, smoking and stroke. In addition,
several evidences highlight the correlation between high alcohol
consumption, saturated fats and cholesterol intake. The
apolipopoteins are cholesterol transporters of high importance for
neuronal plasticity, CNS glucose utilization and mitochondrial
functions. The apoE isoforms (2, 3 and 4) differ in their abilities to
accomplish these critical tasks. Approximately 20% of the
population carries one or two apoE4 alleles. ApoE4’s involvement in
neuropathology is well documented factor for AD in many
populations. In some, 40–80% of patients with AD possess at least
one apoE4 allele. While ApoE3 and apoE2 are effective in
maintaining brain homeostasis, apoE4 seems to have an opposite
rule. Impaired cognition in non-dement individuals carrying the
apoE4 allele worsens with age, suggesting a global detrimental
effect on the CNS. Although apoE4 is strongly linked to AD
pathology, its mode of action is unknown. Insights into the role of
apoE in neuropathology have come from studies of transgenic mice
expressing human apoE3 or apoE4 in neurons or astrocytes.
Features of AD pathology in these animals include reduced numbers
of presynaptic terminals in mice expressing apoE4 with or without
expression of human amyloid precursor protein (APP), increased
plaque deposition in apoE4 mice expressing APP, increased
phosphorylation of tau, impaired learning and memory, and altered
long-term potentiation. No studies are available on the correlation
between life style and apoE4 genotype in AD incidence. Aim of this
project is to study the impact of different life conditions in relation
to the apoE genotype on neurodegeneration. Wild and transgenic
mice expressing human apoE3 or apoE4 are exposed to different
diet conditions such as a normal diet, high saturated fats and
cholesterol intake. After 6 months of diet mice will be tested to
evaluate cognition and memory impairments. In the same subjects,
gene expression (cDNA microarrays) will be studies in discrete brain
areas. RT-PCR assay will be used to confirm the results. Moreover,
selected genes will be further analyzed with classical biochemical
procedures. In addition, other AD proteins such as Aβ, APP and Tau,
and several markers for apoptosis, necrosis and neurodegeneration,
will be investigated with biochemical methods in order to determine
their quantity, their activity and their distribution in the brain. The
combination of high throughput methods with the classical
biochemical assays will allow a deep understanding of the molecular
mechanisms behind the studied risk factors involvement in
neuropatology in relation with apoE genotype. The obtained results
will provide important information to define life guidelines for
people carrying the apoE4 allele, as prevention for AD and
neurodegenerative disorders. This could be of relevance in public
health strategies. Furthermore, results could help to identify new
therapeutic targets and early diagnostic markers. Moreover, this
new experimental approach will develop a new animal model for AD
that has a more natural onset of the pathology.
Rosa Maria Corbo
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
[email protected]
Dip. Genetica e Biologia Molecolare, Università La Sapienza, Roma
Sporadic Alzheimer’s disease (AD) is a complex neurodegenerative disease caused by
the interaction of genetic and nongenetic factors. The e * 4 allele of the apolipoprotein E gene
(APOE) is the only well-established genetic susceptibility factor for sporadic AD, though
numerous risk alleles in other genes are also thought to be involved in AD development.
Gender is a relevant AD risk determinant and there is evidence for a higher prevalence of AD
in women. However, whether this is due to the longer life expectancy of women or to
biological gender- specific risk factors for the disease is unclear. It has been hypothesized that
a declining estrogen level following menopause and subsequent reduction in its
neuroprotective action may contribute to the development of the disease in women. In line
with this hypothesis, the possible role of estrogen nuclear receptor genes (ESR1 and ESR2) as
AD risk factors has been largely investigated. In addition there is evidence that past fertility is
associated with a higher AD risk and a reduced AAO of the disease, probably because of the
lifelong decrease in estrogen levels that parity seems to produce in women.
The aim of the present investigation is to extend previous investigations (1-3) and in
particular 1) to collect data on the reproductive life of AD women to identify those
parameters which could influence the risk of AD and/or AD onset age; 2) To study the genetic
variability of genes involved in estrogen metabolism or women fertility.
- Corbo RM, Gambina G, Ruggeri M, Scacchi R. Association of estrogen receptor α (ESR1)
PvuII and XbaI polymorphisms with sporadic Alzheimer’s disease and their effect on
apolipoprotein E concentrations. Dement Geriatr Cogn Disord, 22:67-72, 2006- Combined effect of Apolipoprotein E (APOE) genotype and past fertility on age at onset of
Alzheimer’s disease in women. Dement Geriatr Cogn Disord, 24: 82-85, 2007
- Genetic variation of CYP19 (aromatase) gene influences age at onset of Alzheimer’s
disease in women. Dement Geriatr Cogn Disord. 2009, 27, 513-518.
Area di interesse identificata
Genetic susceptibility to Alzheimer’s disease
Alzheimer’s disease in women: AD as a gender disease
A.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Basic research
Investigation on common and rare variation of genes involved in sporadic
Alzheimer’s Disease susceptibility
Università di Roma La Sapienza
5 years
At present the genetic component of the sporadic
late-onset form of Alzheimer disease (LOAD) has been only
partially elucidated, since only the apolipoprotein E (APOE)
gene, specifically the e*4 allele, has been reportedly
associated with the disease. Since the APOE polymorphism
alone does not entirely explain the genetic predisposition to
sporadic AD, investigation into whether any variation in
further candidate genes could account for the remaining part
may offer important clues. However, investigations on AD
candidate genes have usually examined only common
polymorphisms in accordance with the hypothesis “common
disease /common variants” . The additional hypothesis
suggesting the presence of a relevant genetic heterogeneity
due to rare alleles (“common disease/rare variants” ) has
been scarcely investigated . The aim of the present study
was 1) to examine the common variation of numerous
candidate genes (ACT, PSEN1, LDL-R, APOC1, alfa2M, PON1,
LPL, AChE, BChE, Chat, CYP19); 2) to carry out a mutation
screening in the exons of APOE, PSEN1, APP genes by means
of DHPLC (denaturing high performance liquid
chromatography). (product: 12 publication, see pubmed)
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Prof. LUCIA MIGLIORE
University of Pisa
Department of Human and Environmental Sciences
Via S. Giuseppe, 22
56026 PISA Italy
E-mail: [email protected]
Fax: +39 050 2211034
Tel: +39 050 2211029-28
- Genetic susceptibility to Alzheimer’s disease
(Epigenetic marks of susceptibility to Alzheimer disease)
Abstract
AD is characterized by high Hcy and low folate blood levels. Many of the
genes participating in folate metabolism are polymorphic, meaning that
this pathway can be altered by low dietary folate intake, by the
polymorphic variants and/or by combinations of both. In vitro studies
showed that in conditions of altered folate metabolism the status of
methylation of the promoter of the PSEN1 gene underwent a variation,
causing a deregulation of presenilin 1, BACE and APP proteins.
The study is designed to evaluate the contribution of folate metabolism
(including polymorphisms of metabolic genes and plasma levels of folate,
homocysteine (Hcy) and VitB12) in both early (MCI) and late stages of
dementia, and its possible contribution to the methylation status of the
promoter of genes that participate in Aβ processing. The study will be
performed in blood samples with the goal of identifying easily detectable
epigenetic biomarkers of disease susceptibility.
Finanziamenti ricevuti
Titolo progetto
A PILOT STUDY ON THE EFFECTS OF FPP ON THE COGNITIVE FUNCTION
AND ON OXIDATIVE STRESS PERIPHERAL BIOMARKERS IN
MILDCOGNITIVE IMPAIRMENT PATIENTS (MCI)
Ente finanziatore
Durata progetto
Abstract del progetto
OSATO Research Institute, Gifu, Japan
Project ongoing
30+30 amnesic MCI patients divided into two main groups. One treated
with fermented papaya preparation (FPP), the other without treatment
(placebo: PLC).
The study will be performed within 12 months for a total treatment time
of six months. After six and twelve months a report will analyze the
results. At the beginning of the pilot study all the subjects recruited will
be screened for the Total Antioxidant Status (TAS) that will include the
determination of the following values in blood: selenium, folic acid,
vitamin B12, C, E together with the evaluation of the GSH/GSSG ratio. The
groups will be analyzed for assessing primary and oxidative DNA damage
levels (Comet assay) and chromosome damage also due to oxidative
damage (Cyt-B micronucleus assay) in peripheral blood leukocytes.
Moreover 8OH-dG levels in urines samples will be evaluated.
We plan also to evaluate the following biochemical parameters of
oxidative stress: advanced oxidation protein products (AOPP);
glutathione; hydroxynonenal; lipoperoxide (basal and under stress
conditions); lactate (basal and under stress conditions); mitochondrial
metabolite (by Mass Spectroscopy).
The decrease (if any) of oxidative stress biomarkers due to the FPP
subministration will be evaluated by comparing after12 months the two
MCI groups (FPP treated and placebo controls) data by means of
statistical analyses. At the same time the cognitive impairment evolution
will be monitored by means of the neuropsychological evaluations also
within the treatment period (0, 6 and 12 months). Moreover for all the
subjects involved, DNA extraction will be performed and the samples will
be stored for further genotype analysis (e.g.: ApoE, ..).
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse
identificata
Finanziamenti ricevuti
Titolo progetto
LAURA COLOMBAIONI
[email protected]>
Istituto di Neuroscienze CNR, Pisa
Role of altered lipid and cholesterol metabolism in neurodegenerations
Ente finanziatore
Associazione Italiana Niemann-Pick
Durata progetto
3 years
Abstract del progetto
Deregulated lipid metabolism is of particular importance in pathogenesis of CNS disorders
and injuries. For this reason the role of cholesterol and lipid metabolism as susceptibility
factors in neurodegenerations needs to be further investigated. Cholesterol is an
important regulator of lipid organization in neuronal membranes and is the precursor for
neurosteroid biosynthesis. Low levels of neurosteroids are related to poor outcome in
many brain pathologies. The deregulation of the principal cholesterol carrier protein in
the brain, the Apolipoprotein E, is a significant risk factor for Alzheimer's disease.
Similarly, Parkinson's disease is, in some degree, caused by an enhancement of lipid
peroxidation. Also Niemann-Pick A-B diseases are caused by sphingomyelin accumulation,
while Niemann-Pick disease C is due to mutations in either the NPC1 or NPC2 genes,
resulting in defective cholesterol and sphingolipid transport. Sphingolipid metabolites are
emerging as members of a novel class of lipid mediators, acting both as intracellular
second messengers and as ligands of cell surface receptors, capable of controlling
neuronal proliferation, differentiation and apoptosis.
In my laboratory, the effects, on neuronal cells, of an altered lipid
metabolism are examined. The goal is to understand how the altered lipid
metabolism produces neuronal injury, what subcellular compartments are
involved and how this occurs. An interdisciplinary approach is used employing
cell biology, molecular biology and live cell imaging methods. By using confocal
microscopy, some key events associated to altered cholesterol trafficking or to
sphingolipid unbalance are characterized, such as mitochondrial deregulation,
release of pro-apoptotic effectors and alterations of calcium homeostasis in
specific subcellular compartments.
Cellular mechanisms of neuronal dysfunction due to altered lipid metabolism.
In particular, the effects on neuronal calcium homeostasis of some sphingolipid
metabolites such as Glucosyl-ceramide Sphingosine and Sphingosine 1-phosphate
are analyzed. Our results indicate that the calcium balance in mitochondria and
in Endoplasmic Reticulum is regulated by cholesterol and sphingolipid
metabolites and that this is critically involved in control of neuronal physiology.
Understanding the impact of lipid metabolism changes in neuronal cell functions will
promote novel concepts and will reveal new therapeutic targets for a variety of nervous
system disorders.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Roberto Cosimo Melcangi
02-50318238
[email protected]
DEFIB-Università degli Studi di Milano
F. Endocrine and metabolic factors
Sex-specific therapeutic strategies based on neuroactive steroids for
Alzheimer's disease.
partial support by Bayer-Pharma
2 years
The importance of sex in the incidence of Alzheimer's disease (AD) has
been studied. Indeed, epidemiological studies support a higher
prevalence and incidence of AD in women. Moreover, as observed in
several experimental models and in AD patients, parameters such as
oxidative damage, plaque load, amyloid precursor protein, β-amyloid
production,
seizure
activity,
locomotion,
hippocampal
neurodegeneration, astrocyte proliferation, level of brain-derived
neurotrophic factor, cognition are differently affected in a sex-dimorphic
way. Although sex differences in the brain may be in part the
consequence of the genetic complement of the cell, differentiation of the
neural tissue is also influenced by the developmental actions of sex
hormones. Both genetic sex and developmental actions of sex hormones
contribute to the generation of sexually dimorphic neuronal networks in
the brain and the spinal cord, and both factors may influence the
outcome of pathological insults to the nervous system. Thus, some sex
differences in brain pathology may be the consequence of the functional
and morphological differences in neural structure between the sexes.
However, it is important to remember that CNS is not only a target for
steroid hormones coming from peripheral glands but also of those
directly synthesize in loco (i.e., neurosteroids). Moreover, steroid
hormones are also actively CNS converted into metabolites that are more
active and in some cases utilize a different mechanism of action. Both
steroid hormones and neurosteroids may be included in the neuroactive
steroid family.
Therefore, the assessment of the levels of neuroactive steroids directly in
male and female brain areas of experimental models of AD might be
extremely important. Using our previous experience in other
experimental models of neurodegeneration, we intend to evaluate the
levels of neuroactive steroids by liquid chromatography tandem mass
spectrometry and to confirm possible changes analyzing by real time PCR
the gene expression of molecules and enzymes involved in their
synthesis. These observations might be very useful to identify possible
pharmacological target useful to design sex-specific therapeutic
approaches based on neuroactive steroids.
Nome
Contatti
Tel.
E mail
Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Roberto Maggi
02 50318233 – 3293718540
[email protected]
Endocrinology, Physiopatology, Applied Biology
Selective Alzheimer Diserase Indicator-1 (Seladin-1): a linker between
cholesterol, oxidative stress and Alzheimer’s disease.
PRIN – MIUR 2006069900_002
2 anni
Alzheimer's disease: correlation with the Gonadotropin Hormone
Releasing Hormone (GnRH).
Amyloid beta (Abeta) plaques represent one of the primary pathological
features that appear in the brains of Alzheimer's patients. Recent findings
suggest that the complete proteolytic machinery required for Abeta
generation is located within lipid rafts. Membrane cholesterol content is
crucial for lipid raft organization and it seems to regulate the membrane
localization and the activity of secretases involved in the metabolism of
APP. Epidemiological and biochemical studies indicate that all changes in
hypothalamic-pituitary gonadal (HPG) hormones (sex steroids, LH,
Gonadotropin releasing Hormone or GnRH) levels following
menopause/andropause are involved in the cognitive and
neuropathological changes observed in AD. Sex steroids have been shown
to be potent neuroprotective agents and a decreased incidence and delay
in the onset of AD has been described among women on hormone
replacement therapy (HRT).
Direct roles of GnRH in AD have received scant attention, even if phase III
clinical trials are currently underway to establish the role of a GnRH
agonist in lowering the incidence of dementia and in improving cognitive
functions of AD patients.
Recently, it has been discovered that estrogens are able to up-regulate
the new gene seladin-1 (Selective Alzheimer disease indicator 1). Seladin1 is a gene down-regulated in the brain areas involved in AD and its overexpression confer protection against Abeta mediated toxicity. Seladin-1
was found to be identical to DHCR24, the enzyme that catalyzes the last
step of cholesterol biosynthesis. Therefore seladin-1 could represent a
key regulator of membrane cholesterol and it could be the molecular
mediator of the neuroprotective effect of estrogens. This project is
devoted to study the role of two hormones of the HPG axis, estrogen and
GnRH, in AD pathogenesis. Moreover, we will investigate the molecular
mechanisms that can link these hormonal actions with the cholesterol
content of plasma membrane and with the cholesterol-rich microdomains
organization and functionality, and finally the possible role for seladin-1
as a molecular mediator of this biological mechanisms. For this research
proposal we will utilize different appropriate humane (FNC-B4 and SHSY5Y) and murine (GT1-7, GN11, and primary hippocampal neurons)
cellular models.
The experimental plan aims to clarify three points:
1.The involvement of membrane cholesterol and lipid membrane
organization in the processing of the APP and its modulation by
estrogens. We will understand if estrogen stimulation is able to alter the
organization of membrane lipids and proteins and the proteolytic
processing of the APP, and if these modifications correlate to changes of
the cholesterol content and of seladin-1 expression.
2.The role of GnRH in the modulation of the cytotoxic action of Abeta and
the APP processing and in neuronal excitability. We will clarify how GnRH
affect APP processing, Abeta deposition and Abeta toxicity and how GnRH
modulate excitability in hippocampal neurons.
3.The hormonal regulation of seladin-1, its ability to modulate the HPG
hormone action and its involvement in Abeta induced cytotoxicity. We
will evaluate the role of seladin-1, as a regulator of membrane
cholesterol, in the amyloidogenic pathway, in the amyloid-induced
neurotoxicity and in lipid rafts organization and functionality as "signaling
platforms".
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
PRIN – MIUR 2006069900_002
PUR – MIUR 2008
2 anni
Alzheimer's disease: correlation with the Gonadotropin Hormone
Releasing Hormone (GnRH).
Amyloid beta (Abeta) plaques represent one of the primary pathological
features that appear in the brains of Alzheimer's patients. Recent findings
suggest that the complete proteolytic machinery required for Abeta
generation is located within lipid rafts. Membrane cholesterol content is
crucial for lipid raft organization and it seems to regulate the membrane
localization and the activity of secretases involved in the metabolism of
APP. Epidemiological and biochemical studies indicate that all changes in
hypothalamic-pituitary gonadal (HPG) hormones (sex steroids, LH,
Gonadotropin releasing Hormone or GnRH) levels following
menopause/andropause are involved in the cognitive and
neuropathological changes observed in AD. Sex steroids have been shown
to be potent neuroprotective agents and a decreased incidence and delay
in the onset of AD has been described among women on hormone
replacement therapy (HRT).
Direct roles of GnRH in AD have received scant attention, even if phase III
clinical trials are currently underway to establish the role of a GnRH
agonist in lowering the incidence of dementia and in improving cognitive
functions of AD patients.
Recently, it has been discovered that estrogens are able to up-regulate
the new gene seladin-1 (Selective Alzheimer disease indicator 1). Seladin1 is a gene down-regulated in the brain areas involved in AD and its overexpression confer protection against Abeta mediated toxicity. Seladin-1
was found to be identical to DHCR24, the enzyme that catalyzes the last
step of cholesterol biosynthesis. Therefore seladin-1 could represent a
key regulator of membrane cholesterol and it could be the molecular
mediator of the neuroprotective effect of estrogens. This project is
devoted to study the role of two hormones of the HPG axis, estrogen and
GnRH, in AD pathogenesis. Moreover, we will investigate the molecular
mechanisms that can link these hormonal actions with the cholesterol
content of plasma membrane and with the cholesterol-rich microdomains
organization and functionality, and finally the possible role for seladin-1
as a molecular mediator of this biological mechanisms. For this research
proposal we will utilize different appropriate humane (FNC-B4 and SHSY5Y) and murine (GT1-7, GN11, and primary hippocampal neurons)
cellular models.
The experimental plan aims to clarify three points:
1.The involvement of membrane cholesterol and lipid membrane
organization in the processing of the APP and its modulation by
estrogens. We will understand if estrogen stimulation is able to alter the
organization of membrane lipids and proteins and the proteolytic
processing of the APP, and if these modifications correlate to changes of
the cholesterol content and of seladin-1 expression.
2.The role of GnRH in the modulation of the cytotoxic action of Abeta and
the APP processing and in neuronal excitability. We will clarify how GnRH
affect APP processing, Abeta deposition and Abeta toxicity and how GnRH
modulate excitability in hippocampal neurons.
3.The hormonal regulation of seladin-1, its ability to modulate the HPG
hormone action and its involvement in Abeta induced cytotoxicity. We
will evaluate the role of seladin-1, as a regulator of membrane
cholesterol, in the amyloidogenic pathway, in the amyloid-induced
neurotoxicity and in lipid rafts organization and functionality as "signaling
platforms".
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Laboratory of Biochemistry & Molecular Biology of Lipids - Mass
Spectrometry "Giovanni Galli"
Donatella Caruso, Maurizio Crestani, Emma De Fabiani, Nico Mitro
0250318344 – 0250318393
[email protected], [email protected],
[email protected], [email protected].
Department of Pharmacological Sciences
Nuclear receptors, coregulators, neuroactive steroids and epigenetics in
neurodegenerative diseases associated to metabolic disorders.
1. MECHANISMS OF NEURODEGENERATION
G. Cell interactions
Nome
Contatti
Istituto/Dipartimento
Prof. Ubaldo ARMATO, MD
[email protected]
Histology & Embryology Unit, Dept. of Life and Reproduction Sciences,
University of Verona Medical School, Verona, Venetia, I-37134, Italy
(in collaboration with Balu Chakravarthy e James F. Whitfield, Institute
for Biological Sciences, National Research Council of Canada, Ottawa,
Ont., K1A 0N6, Canada)
Proposta di ricerca
The Astrocyte, not just a Bystander in Alzheimer’s Disease
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
-
Multidisciplinary projects
Basic research
-------The Astrocyte, not just a Bystander in Alzheimer’s Disease
There are two kinds of Alzheimer’s disease (AD). There is the rare early
onset AD (EOAD) which includes less than 5% of the cases and is due to
mutation-induced hyperproduction of the Aβ42 (amyloid β-(1-42)
fragment of the membrane-associated amyloid precursor protein (APP).
Then there is the slowly developing, late-onset AD (LOADAD that includes
95-99% of the cases. The mutant driver genes of EOAD have been
relatively easy to identify. But the key LOAD drivers have been far harder
to find in the host of changes associated with the progressive breakdown
of neuronal-glial circuits starting in the entorhinal cortex and spreading
through the hippocampus and into the cerebral hot-spots of the cerebral
DMN (default mode neuronal network ) ending in catastrophic cognitive
failure with the loss of memory formation by the hippocampus being an
outstanding feature. So far the attention of workers in the AD field
focused on neurons.
But things are changing. Now we know that certain (radial) astrocytes are
the stem cells for adult neurogenesis in the hippocampal dentate gyrus
and therefore are key components of the memory-forming machinery.
We are learning that massive astrocyte syncytial networks connect to,
and collaborate with, neurons in sustaining and driving brain functions.
And they are co-conspirators with neurons in the cognitive catastrophe of
AD. Therefore we have developed a method of culturing normally
functioning (minimally altered) astrocytes directly following their removal
from adult human cerebral cortices. We are using these cells, which we
call NAHAs (normal human astrocytes), to explore the possible roles of
astrocytes in AD. There is an ever-escalating list of things that might cause
of help cause LOAD. At first it was thought that the striking fibrillar
amyloid plaques in AD brains were the neuronal circuit breaking killers.
But plaque-rich brains of cognitively normal or even above-normal 90-
years old nuns and the failure of plaque formation in transgenic mice to
correlate with neuronal and cognitive loss argue that this is probably not
true. It is now suspected that it is a build-up of invisible pre-plaque Aβ42
oligomers that start destroying synapses and disconnecting cognitive
circuits. We have been and will keep working on three things that seem
to be part of the LOAD process: the induction of VEGF production by
astrocytes; the expression and function of p75NTR, an Aβ42 and
neurotropin receptor.
VEGF Production We have so far found that both Aβ42 and a
combination of three glial-induced cytokines (IL-1β, IFN-γ and TNF-α) that
appear in the inflammatory environment of an AD brain can induce
NAHAs to produce and secrete mainlythe VEGF-A165 isoform. Interestingly
none of the three cytokines can significantly stimulate VEGF production,
but together they are more potent stimulators than Aβ35-42 (an active
fragment of Aβ42). We must now find out how Aβ42 (i.e., Aβ35-42) by
itself and how the three cytokines together produce the HIF-1α•HIF-1β
heterodimer that stimulates VEGF production in the NAHAs. Is VEGF just a
minor player in AD? What might VEGF do in an AD brain? The release of
angiogenic VEGF from the Aβ42/cytokine ensemble-stimulated astrocytes of
the neurobarrier-coupling units with their end-feet attached to blood vessels
damaged by deposited Aβ42 would be expected to reverse, or at least
minimize, the Aβ42-induced vascular damage widely believed to be
responsible for hypoxia, glucose shortages, breached blood-brain barrier and
failing Aβ42 drainage. But then it may not. Hypoxic endothelial cells in the
hypoperfused regions of an AD brain downregulate their MEOX-2 homeobox
gene and its GAX protein product. This causes the endothelial cells to
upregulate the AFX1 forkhead transcription factor that downregulates their
anti-apoptogenesis Bcl-XL gene by stimulating the BCL-6 transcriptional
repressor. The downregulation of MEOX-2 and its GAX protein product also
lowers the level of the Aβ42-clearing LRP1. Therefore while the increased
VEGF production in an AD brain would indeed stimulate angiogenesis, the new
blood vessels would be dysfunctional with endothelial cells unable to clear
Aβ1-42 and hyper-prone to apoptogenesis. But VEGF may have another
impact on memory formation. It is a major factor in the maintenance of the
vascular-based the radial glial (astrocyte-like) neuronal stem cell niche in the
dentate gyral subgranular zone, a center of adult neurogenesis. VEGF
stimulates the production of the niche’s endothelial cells that produce the
neurostimulatory BDNF factor. Thus, angiogenesis is linked to neurogenesis
in the niche to continuously supply the new granular cells needed to start
the recording the memory of novel polymodal inputs from the entorhinal
cortex. Therefore the increased expression of VEGF in a normal brain or
injection of VEGF into such a brain might promote the survival and
proliferation of neural progenitor cells as indicated by the reported
response of hippocampal neuronal progenitor cells to infusion of the
protein into the lateral ventricles of adult rats. But this may be unlikely in an
AD brain even with its elevated VEGF expression, because it has been
reported that the core AD pathology prevents any VEGF-stimulated
immature granule cells from differentiating into mature granule cells.
p75NTR, the villainous Darth Vader of the AD brain? p75NTR combines with
the TrkA neurotrophin receptor to drive brain development when activated
by BDNF and NGF. But it fades away after development only to re-emerge
in the aging, Aβ42 –accumulating AD brain along with the downregulation
of TrkA. Now without restraint by TrkA it becomes dangerous because it
has a cytoplasmic apoptogenic DD (‘death domain’) which is turned on
when the receptor is activated by Aβ42 binding to its neurotrophin binding
site. When its DD is not opposed by associated by TrkA and it meets Aβ42
on a neuronal surface it kills the cell via its DD. We have found that
accumulating Aβ42 actually stimulates the expression of p75NTR by
neurons and NAHAs. Obviously this could accelerate AD development.
Aβ42 and neurotropin receptor. But then there are also indications of a
Aβ42/ p75NTR vicious cycle. Activated p75NTR receptors have been shown
to stimulate caveolar sphingomyelinase to release ceramide from a
pool of sphingomyelin in the cell membrane. The released ceramide
post-transcript-ionally stabilizes BACE1 which, by β-cleaving membraneassociated AβPP, accelerates the drive to full-blown AD by increasing Aβ42
production. This Aβ42would then switch back and bind to p75NTR receptors
and increase the calpain-driven cleavage of the receptor’s C-terminal
membrane-associated domains which ultimately induce the receptors’
cytoplasmic death domains and trigger a cytocidal apoptogenic caspase
8/caspase 3 cascade. We are now trying to confirm the contrapuntal rise
and fall of p75NTR and TrkA in a key region of the human AD brain such as
the hippocampus. And along with this we are trying to find out with
whether NAHAs can start an Aβ42/ p75NTR vicious cycle. If they can, it will be
a strong indication that astrocytes have a major role in the cognitive
collapse of the AD brain.
Nome
Contatti
Manuela MARCOLI
Dipartimento di Medicina Sperimentale (DIMES), Sezione di Farmacologia
e Tossicologia – Università degli Studi di Genova
Viale Cembrano, 4 - I-16148 Genova, Italy
Phone: +39 010 3532656; Fax: +39 010 3993360
E-mail: [email protected]
Istituto/Dipartimento
Dipartimento di Medicina Sperimentale (DIMES), Sezione di Farmacologia
e Tossicologia – Università degli Studi di Genova
The research strategy we propose addresses the need for better knowledge of early molecular events
leading to neuronal dysfunction in Alzheimer’s disease (AD). We propose to focus on the effects of soluble
amyloid-beta peptides (Aβ) on mechanisms sustaining neuron-astrocyte cross-talk at glutamatergic
synapses and on network synaptic activity in Multi Electrode Array (MEA)-coupled neuron/astrocyte
cultures, in both wild type and triple transgenic AD mouse (3xTg-AD mouse). Combined use of isolated
models (nerve terminals and astrocyte processes) and an integrated model (MEA-coupled
neuron/astrocyte cultures) will provide complementary data to understand Aβ-related dysruption of
glutamatergic transmission in AD. Assessment of changes of synaptic transmission at the level of astrocyte
processes and nerve terminals (exposed to Aβ) together with analysis of the effects of (endogenously
produced and exogenously added) Aβ on neuron/astrocyte network activity may shed new light on the
molecular and cellular mechanisms underlying early dysfunction of glutamatergic transmission in AD,
helping to discover new therapeutic targets to improve nerve cell signal transduction and ultimately
preventing neuronal dysfunction.
Background and rationale: Astrocytes regulate neuron function through nonoverlapping domain
organization: one astrocyte processes contact thousands of synapses, promoting neuronal synchrony and
regulating synaptic transmission; the neuro/gliotransmitter glutamate mediate bidirectional neuronastrocyte communication. Although altered glutamate transmission and astrocyte global perturbation are
reported in AD, scarce attention is focused on the potential role of astrocyte processes in synaptic
dysfunction in AD. Soluble Aβ appears involved in synaptic dysfunction; focus on the impact of Aβ on
mechanisms sustaining neuron-astrocyte cross-talk at glutamatergic synapses might unravel key steps in
synaptic impairment and in the cascade of events causing specific sets of neurons becoming dysfunctional.
Experimental models and Methods: Aβ effects on [Ca2+]i and transmitter release will be directly
investigated on astrocyte processes (gliosomes) and nerve terminals (synaptosomes) from adult wild type
and 3xTg-AD mouse brains. Gliosomes and synaptosomes stem from astrocytes and neurons obtained
from adult mouse brain; in situ maturation of neurons and astrocytes would assure that the impact of
intracellularly accumulating Aβ would also be accounted for when studying glutamate release and Ca2+
signaling in nerve terminals and astrocyte processes prepared from 3xTg-AD mouse brain. Integrated
effects of Aβ on network activity and synaptic connection maturation will be analyzed by non-conventional
electrophysiology on cultured neurons/astrocytes from wild type and 3xTg-AD mouse brain on MEAs,
allowing long-term, non invasive, multielectrode detection of neuronal network activity and
synchronization; the bioelectronic system has proven very sensitive in detecting early neuron damage and
loss of synaptic transmission.
Human resources: key persons, experience and know-how that can be accessed without additional funds:
Manuela Marcoli, Guido Maura (University of Genova, Department of Experimental Medicine DIMES;
Center of Excellence for Biomedical Research CEBR; National Institute of Neuroscience INN): transmitter
release from CNS preparations (nerve terminals, astrocyte processes) and cell cultures
Sergio Martinoia (University of Genoa, Department of Biophysical and Electronic Engineering DIBE;
Italian Institute of Technology IIT): Micro electrode arrays technology; MEA-based systems and in-vitro
neural interface and neuro-robotics
Mario Nobile (National Research Council, Institute of Biophysics, Genova): [Ca2+]I imaging on single nerve
terminals and astrocyte processes
Area di interesse identificata First challenge: scientific; understanding the mechanisms of the disease
Priorites: - Basic research
- Multidisciplinary projects
- New treatment strategies
Finanziamenti ricevuti - ongoing research support
Titolo progetto
Neuronal differentiation: functional evaluation of neurotransmission
Ente finanziatore
MIUR – PRIN mod B Anno 2008 - prot. 20088JEHW3_005
(to Guido Maura)
Durata progetto
2 years
Abstract del progetto
To understand the molecular mechanisms associated to the
differentiation of neuron precursor cells, we plan to use Neuroblastoma
N18TG2 clones, transfected with Choline Acetyl Transferase (ChAT) and
overexpressing early growth response factor-1 (EGR-1), that mimic
neuron precursor cells. The research program plans to investigate on the
possibility to induce a fully developed neuron phenotype in N18TG2
clones transfected with ChAT and/or overexpressing EGR-1, by inhibiting
the expression of the Repressor element-1 Silencing Transcription (REST),
coding for a transcriptional repressor of neurogenesis. Our analyses will
focus on evaluation of the cell ability to communicate through
neurotransmitters and regulated exocytosis upon REST knock down; if
this occurs the gene expression profile will be determined. The project
will reveal gene networks relevant for complete maturation of the
neuroblastoma cell line, and hopefully will be of help in understanding
the neuron maturation process.
Finanziamenti ricevuti - ongoing research support
Titolo progetto
Mechanisms of glial and neuronal glutamate release as specific
therapeutic targets in brain ischemia
Ente finanziatore
Fondazione Cassa di Risparmio di Genova, 2008
(to Mario Nobile, role of MM: participant)
Durata progetto
2 years
Abstract del progetto
The aim of the project is to get new light on molecular and cellular
mechanisms activated during/after ischemia to find out pharmacological
targets for neuroprotection. Brain ischemia results from multiple related
mechanisms involving both neurons and glial cells. The neuron-glia crosstalk, subserved by mechanisms including glutamate is of fundamental
importance to the synapse physiology and to the synaptic transmission
dysregulation in neurodegenerative diseases; increased extracellular
glutamate and intracellular calcium overload lead to cell death. Multiple
mechanisms are responsible for glutamate release: vesicular release,
glutamate transporter reversal, and possibly activation of ATP
P2X7receptors, of volume-regulated anion channels or of
connexin/pannexin. hemichannels. These ways for glutamate release can
function both on neurons and astrocytes, although the role for glial cells
in ischemic neuron damage is still debated. As the pharmacological
intervention based on glutamate ionotropic receptor antagonism was
unsatisfactory, knowledge of the glutamate modes of exit during ischemia
might help to find new specific therapeutic targets. The research strategy
will be based on the investigation at synapse level, at the level of single
cell and of the cell network (slices), on the participation of P2X7
receptors, VRAC channels and pannexin in ischemic damage in vitro. As
we previously found that adenosine controls astrocyte P2X7 receptors,
we will investigate on the relationship between adenosine and P2X7
receptors during ischemic insult.
Finanziamenti ricevuti - submitted proposal
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Synapse and Amyloid-beta: effects on astrocyte processes and nerve
terminal
Alzheimer’s Association 2010 Investigator-Initiated Research Grant
Proposal Identifiers LOIID: 173512: approved
Full proposal: under evaluation
(role of MM: principal investigator)
3 years
Understanding of neuron-astrocyte interaction at synapses is crucial to
successful therapeutic approach to neurodegenerative diseases.
Astrocytes regulate neuronal function through nonoverlapping domain
organization: the processes of one astrocyte contact thousands of
synapses, promoting neuronal synchrony and regulating neuron
excitability and synaptic transmission. The neuro/glio transmitters
glutamate and ATP mediate bidirectional neuron-astrocyte
communication at synapses. Although altered glutamate or ATP
transmission and astrocyte global perturbation are reported in AD
models, scarce attention has been focused on the potential role of
astrocyte processes in synaptic dysfunction in AD. Soluble amyloid-beta
peptides are involved in synaptic dysfunction, an early event in the
pathogenesis of AD before synapse loss. Our proposal focus on the effects
of soluble amyloid-beta on the mechanisms subserving bidirectional
communication between astrocyte processes and nerve terminals. Focus
on the impact of amyloid-beta peptides on astrocyte process might
unravel key steps in synaptic impairment and in the cascade of events
causing specific sets of neurons becoming dysfunctional. How? Astrocyte
processes (gliosomes) and nerve terminals (synaptosomes) from wild type
and transgenic AD mouse brains will be used, allowing direct investigation
on the amyloid-beta effects on intracellular calcium and transmitter
release. It is worth noting that gliosomes and synaptosomes stem from
astrocytes and neurons obtained from adult mouse brain; in situ
maturation of gliosome-producing astrocytes and of synaptosomeproducing neurons provides experimental models suitable to investigate
on amyloid-beta effects in adult life. Integrated effects of amyloid-beta on
network activity and synaptic connection maturation will be analyzed on
co-cultured neurons/astrocytes from wild type and transgenic AD mouse
brain on multielectrode arrays, by non-conventional electrophysiology
(allowing long-term, non invasive, multielectrode detection of
spontaneous and evoked firing activity and neuronal network
synchronization). This bioelectronic system has proven very sensitive in
detecting early neuronal damage and loss of synaptic transmission. Why?
Assessment of changes of synaptic transmission at the level of astrocyte
processes and nerve terminals (exposed to amyloid-beta) together with
analysis of the amyloid-beta effects on neuron/astrocyte network activity
may shed new light on the molecular and cellular mechanisms underlying
early neurodegenerative processes and cognitive dysfunction in AD,
helping to discover new therapeutic targets to improve nerve cell signal
transduction and ultimately preventing neuronal dysfunction and synapse
loss.
Finanziamenti ricevuti - completed research support
Titolo progetto
Feasibility of a prototypical system development for in vitro testing for
biomedical, toxicological and ambiental purpose
Ente finanziatore
Scientific and Technological Park of Liguria Program Objective 2 20002006 (to Sergio Martinoia, role of MM: responsible of Unit)
Durata progetto
2 years
Abstract del progetto
The goal of this project was to set up neuron cultures on multielectrode
arrays allowing neuropharmacological/neurotoxicological analysis of the
integrated effects of neuroactive substances on network activity
Combining the study of the network firing activity with the study of the
release of the excitatory transmitter glutamate and of the activation of
the intracellular pathways following ionotropic receptor (NMDA,
AMPA/kainate) activation and involved in the excitotoxic neuron cell
damage or death would allow an integrated mechanistic-based approach
to neurotoxicity evaluatio in neuron network. Non conventional
electrophysiological
and
neurochemical
approach
allow
neuropharmacological /neurotoxicological analysis of the integrated
effects of neuroactive substances on the network activity and dissection
of the mechanisms underlying complex network responses to the
substances.
Finanziamenti ricevuti - completed research support
Titolo progetto
Pharmacological control of extracellular excitatory amino acid
accumulation and cellular calcium overload in neonatal
ischemia/reperfusion
Ente finanziatore
Mariani Foundation (Milan) R-07-66
(to Mario Nobile, role of MM: participant)
Durata progetto
2 years
Abstract del progetto
The study evaluates acute and long-term (60 days) effects of perinatal
hypoxia, in order to find new pharmacological targets to be used in
intervention devoted to prevention/protection against neonatal ischemic
neuron damage. Perinatal hypoxia-ischemia is a major cause of neonatal
morbidity and mortality. A number of evidences indicate that ischemic
neuron damage can depend on increase of extracellular K+ accumulation
of excitatory aminoacids and of other transmitter (such as
adenosine/ATP), and intracellular Ca2+ overload, triggering excitotoxicity
and apoptotic/necrotic nerodegeneration. Such events might play
different roles depending on the developmental stage of central nervous
system. Scarce reports are available on specific models, allowing
dissection of diverse mechanisms possibly involved and temporal
sequences of neonatal ischemia. Relative contributions of neurons and
glia and of their bi-directional communication in the control of glutamate
synaptic level and in excitotoxicity triggering and execution, are not
clearly defined. Knowledge of the modes for efflux of excitatory
aminoacid and their regulation, in early and late phases of
ischemia/reperfusion, could open the possibility to reduce the
extracellular excitatory aminoacid levels in different neonatal ischemia
phases. As intracellular Ca2+ accumulation seems prerequisite for neuron
damage cascade, dampening of Ca2+ influx through ionotropic glutamate
or purinergic receptors (e.g. P2X7) could significantly reduce neuron
damage. We therefore propose investigation on glutamate efflux and
cellular Ca2+ levels in physiological conditions and during ischemia in vitro
models of neonatal brain (preparations from neonatal and 60 days-old
rats, models for premature infant and adulthood brain, respectively).
Parallel studies will be conduced with the aim to unravel whether
glutamate release from astroglial cells can be modulated by endogenous
signaling molecules through the regulation of swelling-activated Clchannels. New pharmacological targets to be used in intervention
devoted to prevention/protection against neonatal ischemic neuron
damage, will be searched for through the following intermediate steps: better knowledge of the functional role played by ionotropic glutamate
and purine receptors on neuron and glial cells; - better knowledge of the
modes responsible for extracellular glutamate accumulation and of their
relative importance in the temporal sequence of events during
ischemia/reperfusion; -better knowledge of the mechanisms regulating
activity of swelling-activated Cl- channels involved in the pathologic
release of excitatory amino acids; - identification of receptors whose
activation (or blockade) might affect vesicular excitatory aminoacid efflux
or glutamatergic pathways or neuron-glia activation in different phases of
ischemia/reperfusion.
Finanziamenti ricevuti - completed research support
Titolo progetto
Neuron networks coupled in vitro to microtransducers and artificial
body: a new potent device to study neuronal interfaces and cerebral
plasticity in vitro
Ente finanziatore
Durata progetto
Abstract del progetto
Italian Ministry of University and Scientific Research (FIRB 2002)
RBAU012KF8_005
(to Guido Maura, role of MM: participant)
3 years
The goal of the project was to investigate on spontaneous and
electrically-evoked activity of neuronal networks on microtransducer
arrays for a better knowledge of the cell mechanisms underlying burst
generation and propagation. To this end, non conventional
electrophysiology on MEA will be applied to detect the effects of
chemical interference on neuron-specific features linked to neuron
functioning (electrophysiological events at level of single cells and at
the network level) and will be combined with neurochemical analysis
for neuron-specific features linked to neuron functioning
(neurotransmitter processing and release; presence and functioning of
neurotransmitter receptors and linkage to transduction mechanisms)
combined with the electrophysiological analysis.
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Domenici Luciano
[email protected]; [email protected]
Neuroscience Institute, CNR, Via G. Moruzzi 1, 56100 Pisa; Dep. STB,
University of L’Aquila, L’Aquila.
Alzheimer’s disease, Ageing
Active projects:
1) RAGE, MAP kinases and Abeta induced synaptic dysfunction.
2) From brain ischemia to beta amyloid toxicity in Alzheimer’s
disease; neurobiological mechanisms, new biomarkers and
therapeutical approach.
Submitted projects:
- Vascular triggering of beta amyloid toxicity in AD.
- BDNF and beta amyloid toxicity, building a bridge from
neurobiological mechanisms to treatment of brain disorders
1) American Health Assistance Foundation (AHAF).
2) Regione Toscana, Regional Health Research Program 2009 (first
step approval passed, final approval and funding pending).
1) Two years.
2) Two years.
1. Abstract project AHAF.
Beta amyloid protein (Aβ) is the principal component of senile plaques, a
neurophatological prominent feature of Alzheimer's disease (AD).
Increasing evidence suggests that loss of neuronal function and cognitive
impairment precede the accumulation of Aβ plaques in AD. We
hypothesize that oligomeric Aβ is involved in neuronal dysfunction during
an early phase of AD. We will investigate the mechanisms underlying
oligomeric Aβ induced synaptic dysfunction in entorhinal cortex (EC)
whose impairment contributes to cognitive decline in AD. We will address
three specific aims:
1. To test the hypothesis that Aβ affects cortical synaptic plasticity. The
effects of different concentrations of oligomeric Aβ on synaptic
transmission and efficacy will be investigated in EC slices using
extracellular and single cell recordings.
2. To verify the identity of receptors and intracellular pathways
transducing Aβ signal. Receptor for Advanced Glycation Endproducts
(RAGE) is a cell surface binding site for Aβ. We will investigate the effects
of oligomeric Aβ on synaptic transmission and efficacy in EC slices: i) in
the presence of anti-RAGE antibodies, ii) in the transgenic (Tg) mice with
targeted neuronal expression of the wild type form of RAGE and in RAGEdeficient mice; iii) in the Tg mice with a dominant negative form of RAGE
targeted to neurons (DN-RAGE). Finally, we will investigate the
involvement of different kinases associated with RAGE activation, such as
p38 MAPK, p42-p44 MAPK and JNK.
3. To test the role of neuronal RAGE in vivo using animal models of Aβmediated synaptic dysfunction. We will employ Tg mice overexpressing
mutant human amyloid precursor protein (mAPP); Aβ-rich environment in
Tg mAPP mice induces synaptic dysfunction. We expect that synaptic
dysfunction observed in Tg mAPP mice does not occur in Tg mAPP mice
crossed with Tg DN-RAGE mice. Finally, we will determine whether
increased expression of neuronal RAGE in Tg mAPP mice exaggerates
synaptic dysfunction.
2. Abstract Project Regione Toscana.
Alzheimer’s disease (AD) is a progressive neurodegenerative disorder of
the elderly characterized by senile plaques composed of beta amyloid
(Aβ), neurofibrillary tangles, synaptic loss and impairment of cognitive
functions. AD is the most common cause of dementia followed by
vascular dementia (VaD). The incidence of neurodegenerative disorders in
AD is increased by vascular pathologies such as stroke, clogging arteries
and hypertension inducing brain hypoperfusion and hypoxia. However, a
causal relationship between AD and brain ischemia at the molecular level
has not been established. The present project will contribute to elucidate
whether Aβ/RAGE signalling may be viewed as part of a neural system
that controls synaptic responses in vascular pathology and AD. We are
interested in investigating this issue as many patients with AD also have
hypertension and vascular infarctions. Designing new biomarkers of
neurodegeneration and interfering with Aβ/RAGE signalling is expected to
have high impact to ameliorate cellular dysfunction in AD and vascular
pathology. Patentability of a new diagnostic method of AD based on
biomarkers associated with RAGE activation and deposition of Aβ
represents the goal of our plan.
Nome
Evelina Chieregatti
Contatti
[email protected]
Istituto/Dipartimento
Dept. of Neuroscience and Brain Technologies, IIT, Genoa, Italy
Proposta di ricerca
Synaptic deficits in neurodegenerative diseases
The main lines of our research concern the investigation of the early mechanisms underlying synaptic
dysfunctions in Parkinson’s and Alzheimer diseases. These neurodegenerative diseases are characterized
by the deposition of proteinacious aggregates in the neurons and in the brain tissue.
However it has been shown that most neurons that undergo cell death do not contain aggregates and
that, whether they contain aggregates or not, are similarly affected by morphological dendritic
abnormalities or biochemical changes. It may thus be conceivable that, rather than being caused by the
accumulation of aggregates with neuronal cell bodies, which could be an epiphenomenon,
neurodegenerative diseases may be characterized by dying back mechanisms that begin at the synapse
and lead to axonal degeneration. Both β amyloid (Aβ) and α-synuclein (Syn) are pathogenetic proteins that
upon mutations or increase in their levels may induce alterations in the synaptic function and neuronal cell
death. A key element in the regulation of neurotransmitter release and in the fine tuning of synaptic
efficacy is the neuronal cytoskeleton. Both proteins are implicated in actin dynamics as well as microtubule
stability through their interactions with a microtubule-binding protein, tau. We recently demonstrated a
disrupting effect of the mutated form of Syn on actin dynamics in neurons. We are studying their effects
on the morphology and dynamics of actin microfilaments and intermediate filaments by the use of purified
proteins and neurons derived from knock-out and transgenic mice. The long term goal of this project is to
investigate the effect of the two proteins on cytoskeletal-regulated processes, such as polarization,
regeneration after axotomy and neuronal plasticity. Another project concerns the characterization of the
molecular interactors of Syn that may have a role in neurodegeneration associated with αsynucleopathies.
The intracellular localization of the cleavage of APP by BACE1 has long been a topic of debate. Early
endosome is the first site of APP endocytic sorting, where APP has been shown to co-localize with BACE1.
Altered endosomal structures have been found in AD brains. Our aim is to identify proteins of interest such
as enzymes or molecular motors that may be found in the APP-containing endocytic vesicles.
It is expected that the results will shed light on the role of Syn and Aβ in synaptic function, and on the
molecular interactors mediating their effect, providing cues useful to reach appropriate therapeutic
endpoints based on the modulation of the signalling pathways involved in the pathogenic effect of Syn and
Aβ.
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Macromolecular interactions and Neurodegeneration
The role of intraneuronal amyloid β(Aβ) peptide on the pathogenesis of
the Alzheimer disease: functional and proteomic analysis.
Fondazione CARIPLO – role: Partner
2 years (end 2009)
Alzheimer disease (AD), one of the neurodegenerative diseases of the
nervous system characterized by a chronic process of cell death, is the
most frequent form of dementia, but there is still no therapeutic
intervention capable of preventing, delaying or treating it. Intraneuronal
Aβ accumulation precedes the deposition of amyloid plaques and the
appearance of neurofibrillary tangles, and correlate with the first
appearance of cognitive deficits. The role of intraneuronal Aβ in the AD
pathogenesis is still unclear, but recent data suggest a possible
involvement in cytoskeletal functions and neurotransmitter release. The
goal of this project is to understand how Aβ may interfere with synaptic
function and, by the use of advanced proteomic techniques, to identify
the proteins that interact with Aβ. The results we expect to obtain will
shed light on the pathogenetic mechanisms in AD and will allow the
development of advanced diagnostic and therapeutic protocols.
Titolo progetto
Pathogenetic mechanisms of familial Parkinson’s disease: wt and A30P
α-synucleins affect the structure of microfilaments and intermediate
filaments. Pathways and effects on cytoskeletal dynamics.
Ente finanziatore
Durata progetto
Abstract del progetto
Telethon-Italy – role: PI
3 years (end 2013)
Overall objectives. To understand a physiological role of alpha-synuclein
(Syn) and its role in the pathology of Parkinson's disease (PD) by
comparative studies of wt and A30P Syn mutant interaction with
microfilaments and intermediate filaments. To understand Syn role in
cellular processes that require active remodeling of these two
cytoskeletal elements.
Specific aims. Analysis of the regulation of actin-Syn interaction in living
cells by fluorescence energy transfer (FRET). Analysis of the composition
of actin/A30P Syn aggregates formed during cytoskeleton remodeling.
Analysis of Syn effects on intermediate filaments turnover and on
neuronal regeneration after axotomy. Investigation of the pathway(s)
that mediates the effect of extracellular Syn on actin cytoskeleton
morphology.
Background. Mutations in the Syn gene are responsible for familial PD.
Syn knock-out mice suggest a role of Syn in the regulation of the presynaptic vesicular pools in neurons. Syn has been involved in neuronal
regeneration occurring during differentiation and repair. Interestingly,
actin microfilaments and the intermediate filaments are actively involved
in every one of these processes.
Description of the project. Based on our data demonstrating interaction
between Syn and actin we propose to study the regulation and the
subcellular localization of their interaction by FRET. In neurons in culture
from knock-out and from conditional transgenic mice we plan to
investigate the role of Syn in regeneration after axonal injury. We
propose to investigate the transduction pathway that is activated by
extracellular Syn.
Anticipated output. We expect to identify the specific step at which Syn
function at the pre-synapse and the possible loss of function of the A30P
Syn mutant. These results will be of great relevance in the understanding
of PD pathogenesis.
1. MECHANISMS OF NEURODEGENERATION
H- Intracellular Traffiking
Nome
Contatti
Istituto/Dipartimento
Cecilia Bucci
Tel. +39-0832-298900; Fax +39-0832-298626
e-mail: [email protected]
Department of Biological and Envirnmental Sciences and Technologies
(DiSTeBA), University of Salento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Membrane traffic and neurodegenerative diseases: role of Rab proteins
and their effectors
The laboratory of Applied Biology directed by Cecilia Bucci is focused on
the role of endocytic Rab proteins. Rab proteins are small GTPases that
control membrane traffic in the endocytic and exocytic pathways. Indeed,
they are responsible, through the recruitment of several specific
effectors, for several aspects of membrane transport as vesicle formation,
vesicle movement onto cytoskeletal tracks, vesicle docking and tethering
to the target compartment, and, finally, vesicle fusion with the target
compartment. There are about 70 rab genes in the human genome.
Alterations of membrane traffic, and, in particolar, of Rab proteins or of
their effector proteins, are responsible for many human genetic and
acquired diseases including cancer. In particular, Rab dysfunction has
been linked to several neurodegenerative diseases. For instance
mutations in Rab7 protein cause the Charcot-Marie-Tooth type 2B
disease, a inherited neuropathy of the peripheral nervous system. Late
endocytic dysfunction has been demonstrated in Alzheimer’s disease and
presenilin (a protein mutated in Alzheimer) interacts with the endocytic
Rab11 and with the Rab regulatory protein GDI (GDP-dissociation
inhibitor). In addition, presenilin influences localization and function of at
least other two Rab proteins, Rab6 and Rab8. Rab6 expression is
increased in Alzheimer’s disease brain. Mutations in alpha-synuclein are a
cause of familial Parkinson disease; mutated alphasynuclein interacts
abnormally with Rab3a, Rab5 and Rab8.
These are only some examples that demonstrate the close connection
between membrane traffic (and in particular Rab proteins function) and
neurodegenerative diseases.
We propose therefore to study different aspects of membrane traffic at
the molecular level, with a particular focus on Rab proteins, in order to
understand the molecular mechanism underlying degenerative diseases
as Alzheimer, Parkinson, Charcot-Marie-Tooth, etc. In particular, we
could investigate the role of the proteins found mutated in these diseases
(i.e. presenilin, alpha-synuclein, Rab7, etc.) in membrane traffic, and the
role of their interaction with regulators of membrane traffic.
Molecular basis of Charcot-Marie-Tooth type 2B disease
Telethon Italy
3 years (2009-2012)
The inherited neuropathies of the peripheral nervous system show
clinical and genetic heterogeneity. CharcotMarieTooth type 2B (CMT2B) is
a ulceromutilating neuropathy clinically characterized by prominent
sensory loss, marked distal muscle weakness and wasting, high frequency
of foot ulcers, infections, and amputations of the toes because of
recurrent infections. Four missense mutations in the small GTPase late
endosomal protein Rab7 have been identified in autosomal dominant
ulceromutilating neuropathy families. These missense mutations cause
the CMT2B phenotype and target highly conserved amino acid residues. It
is not known how these mutations affect specifically peripheral neurons
leading to an axonal pathology in CMT2B. This issue is particularly
challenging considering that Rab7 is a ubiquitous protein. This proposal
plans to continue investigating the
molecular mechanism responsible for the development of the disease.
We have previously biochemically characterized the Rab7 mutated
proteins causative of CMT2B in HeLa cells, determining that they are
mechanistically similar. Indeed, we have demonstrated that activated
forms of Rab7 are responsible for the disease.Within this project we plan
to:
i) do functional studies using the mutated proteins on PC12, SH−SY5Y,
Neuro2a cells and neurons ii) screen DRG and motor neuron cDNA
libraries with different methods to search for new Rab7 interacting
proteins, looking in particular at effectors expressed specifically in the
peripheral nervous system. iii) do functional assays to establish the role of
these newly identified Rab7 interacting proteins and to establish if and
how Rab7 mutations affect their function. Understanding how
dysfunction of Rab7 causes CMT2B will be instrumental to gain insights in
the pathogenesis of this group of disorders, and will open the way to
future identification of therapeutic approaches.
Nome
Contatti
Istituto/Dipartimento
Prof. Paolo Macchi, PhD
Phone: +39 0461-883819/883095
Fax: +39 0461-883937
Email: [email protected]
Molecular and Cellular Neurobiology Lab
CIBIO – Centre for Integrative Biology, University of Trento.
Via delle Regole 101
3060 Mattarello, Trento – Italy
WEB: http://www.unitn.it/cibio
Research proposal
“Comparative analysis of the transport and translational RNPs complexes in neurons: an integrate
transcript- and proteomic approach.”
RNA localization is emerging as an important process to restrict certain proteins to specific subcellular
domains and thus spatially control the expression of genes within cells. One key feature of messenger RNA
(mRNA) localization is that it precedes translation. As a consequence, mRNA has to be kept translationally
silent during its transport towards the proper target compartment. Localized mRNAs are packaged into
ribonucleoprotein particles (RNPs), composed of RNAs and proteins. The localized RNAs contain specific
cis-acting sequence elements, which are recognized by proteins, called trans-acting factors. The
importance of RNA subcellular localization in the formation and function of the nervous system is also
gaining increasing acceptance. In highly polarized cells, such as in neurons, a functional
compartmentalization is crucial. RNA localization is not only involved in establishing the structural polarity
of neurons, but is also assumed to play an important role in synaptic plasticity1. It has been estimated that
approximately 200 different mRNAs encoding for proteins important for synaptic plasticity as well as
various housekeeping genes, are found in dendrites of hippocampal neurons2. Interestingly, the loss-offunction of one component of the RNA transport machinery and/or the misexpression of specific mRNAs
causes abnormalities during neuronal development. For example, defects in axonal and dendritic
outgrowth and/or in dendritic spine development have been described. These phenotypes have recently
been correlated with different neurological disorders, such as FXS (fragile X mental retardation syndrome),
SCA (spinocerebellar ataxia) and SMA (spinal muscular atrophy)3. However, the composition of the
transport as well as the translation control machinery is still poorly characterized. Our research focuses on
dissecting the molecular composition of the neuronal transport RNPs under physiological as well as
pathological conditions (e.g. neurodegeneration, brain tumours and epilepsy).
In detail, we are trying to answer the following biological questions:
1
2
Is the local protein synthesis and/or mRNA trafficking altered in neurons undergoing degeneration?
What is the molecular composition of the neuronal RNPs (RNA and protein interactome) in neurons
affected by degeneration compared to normal cells?
The goal of purifying transport RNPs to identify novel components has been previously attempted, but has
not yielded the precise mechanistic details of the process that are sought. Our laboratory has successfully
demonstrated the feasibility of maintaining endogenous transport complexes containing Staufen, Barentsz
and Pumilio proteins, RNA (RIP-chip analysis), and motor proteins in cell-free extracts of brains and
cultured neurons4,5. Using our collection of affinity-purified antibodies to immunoprecipitate localization
RNPs from brain extract fractions, the composition of RNPs isolated from wild type mice will be assessed
and compared with RNPs isolated from animal model for neurological diseases. We will then test the
individual functions of both known (or suspected) and novel identified components of neuronal RNPs in a
variety of in vivo and in vitro assays to characterize their roles in localization and associated mechanisms.
Finally, since the activation of translation of dendritically localized mRNAs upon specific inputs, such as
synaptic activity, allows for the modulation of existing synapses, notably the morphological and functional
changing of dendritic spines,
3 Are there possible drugs able to affect mRNA targeting (and/or turnover) and local protein synthesis
that can be used to block neurodegeneration?
We are currently setting up a high throughput drug screening assay on neuronal cells at CIBIO (University
of Trento). RIP-Chip experiments will be functional for the development of pharmacological assays where
the level of expression of mRNAs identified will be evaluated through the use of a reporter protein
(luciferase). Through a bioluminescence assay we will be able to quantify the effect of each chemical's
inhibitory activity or promoter of the complex proteins/ mRNA-luciferase.
We propose that a multidisciplinary study to underline the mechanisms that lead to correct regulation of
both RNA and RNA-binding proteins will be crucial for a better understanding of the pathogenesis of
neuronal diseases. Interdisciplinary internal collaborations, with other institutions in Italy and abroad have
been already established to take full advantage of different experimental systems and areas of expertise.
1. Kuhl D. & Skehel P. (1998) Curr. Opin. Neurobiol. 8, 600-606.
2. Job C. & Eberwine J. (2001) Nat. Rev. Neuroscie. 2, 889-898.
3. Dahm R. & Macchi P. (2007). Biol. Cell 99, 649-61.
4. Mallardo M. et al., (2003). PNAS 100, 2100-2105
5. Vessey J. et al., (2010). PNAS 107, 3222-3227
Area di interesse identificata Basic research. Neurodegenerative diseases.
Finanziamenti ricevuti
Titolo progetto
Electrophysiological characterization of alpha-synuclein forming channels,
a protein involved in the neurodegeneration of Parkinson's disease
Ente finanziatore
CARITRO Foundation
Durata progetto
2 years
Abstract del progetto
The aggregation of aberrant proteins is involved in the pathogenesis of
neurodegenerative diseases associated with aging but the mechanisms
leading to neuronal death are unknown. The α-synuclein (αS) is the main
component of Lewy bodies, namely fibrillar intraneuronal inclusions
present in all patients with Parkinson's disease (PD), and is strongly
indicated as the cause of the disease. The αS is not structured in a
"native" form while in vitro form amyloid fibrils that are rich in beta
sheet. Nell'αS mutations found in patients with PD promote the
formation of annular and tubular structures of protofibrille. The
protofibrille of αS, but not the monomer or mature fibrils, bind to
structures such as lipid vesicles, planar and cellular membranes. Here,
they assume a new structural conformation similar to the pores formed
by the bacterial toxins that cross the membrane. These "amyloid pores"
may be the cause of neuronal death, through an uncontrolled and
nonspecific permeabilization of the cell membrane. We intend to
investigate in detail the electrophysiological characteristics of the pores
of αS formed in model systems of increasing complexity: in lipid planar
membranes, synaptic vesicles from liposomes and/or mitochondria and
eventually in cultured neurons. In particular an innovative analysis
technique suitable for noise characterization of pores will be developed.
We intend to investigate, with techniques of analysis of noise, EEG from
healthy patients with PD and to highlight similarities with anomalies
recorded in single cells. We believe that this multidisciplinary approach
will allow 'early diagnosis and more precise onset and evolution of PD as
well as of other neurodegenerative diseases.
1. MECHANISMS OF NEURODEGENERATION
I- Animal models
Nome
Contatti
Istituto/Dipartimento
Adalberto MERIGHI
Phone:++390116709118
Fax: ++390112369118
e-mail: [email protected]
Istituto Nazionale di Neuroscienze e Dipartimento di Morfofisiologia
Veterinaria, università degli Studi di Torino
Via Leonardo da Vinci 44
I-10095 GRUGLIASCO (TO) Italy
Proposta di ricerca
Effects of over-expression of the anti-apoptotic proteins BCL-2 and survivin on the endoplasmic
reticulum stress induced by β amyloid: study of neuroprotection on animal models
Area di interesse identificata Basic research
Developing competitive animal models to study Alzheimer’s disease
The etiology and mechanisms responsible for neurodegeneration in Alzheimer’s disease (AD) are
multifaceted, but recent studies indicate that activation of an intrinsic gene-regulated program of cell
death (PCD = programmed cell death) is a common end-point of the various intracellular pathways leading
to neuronal loss. Multiple intracellular pathways activated during PCD, but, among them, apoptosis plays a
major role. The post-natal cerebellar cortex is an ideal model for the study of PCD, because of the massive
apoptosis of the granule cells, a population of cerebellar excitatory interneurons formed by billions of cells
(see for example Lossi L. and Merighi A. Prog Neurobiol. 2003. 69:287-312). β-amyloid (Aβ) is directly
involved in neurodegeneration observed in AD, where neurodegenerative changes are due, at least in part,
to activation of an apoptotic programme (see for example Wisniewski T. Neurobiol Dis. 1997. 4:313-28). In
primary cortical neuron cultures, the toxic effects of synthetic peptides corresponding to specific
sequences of Aβ have been shown to be consequent to release of calcium across ryanodine receptor (RyR)
channels and/or inositol-3-phosphate channels (IP3) on the membrane of the endoplasmic reticulum
(Ferreiro E. Neurobiol Dis. 2006. 23:669-78). We have recently demonstrated that the block of RyR
channels in organotypic cultures obtained from mouse cerebellum interferes with the increase in density
of cerebellar granule cells expressing the BCL-2 protein (one of the most widely characterized antiapoptotic factors in Eukaryotic cells) fused with a fluorescent reporter marker (EYFP = enhanced yellow
fluorescent protein) after KCl depolarization (Lossi et al. Dev Neurobiol. 2009. 69:855-870). These
observations suggest that BCL-2 could modulate the apoptotic program triggered by Aβ as a consequence
of endoplasmic reticulum stress.
Survivin is one member of a family of proteins commonly referred to as apoptosis inhibiting proteins. It is
expressed during mitosis, and in tumors, but also during normal development of neurons in vivo, as
demonstrated in conditional transgenic mice using the Cre-loxP technology (Jiang Y. et al. J Neurosci. 2005.
25:6962-70). In these animals the conditional deletion of the survivin gene at P10.5 leads to massive
apoptosis in several areas of the central nervous system. As a consequence, newborn mutants display a
marked reduction of brain size, with severe multifocal apoptosis affecting the cerebral and cerebellar
cortices, the brainstem , the spinal cord and the retina. Caspase 3 and 9 (the main effector molecules of
apoptosis)activities are increased, whereas expression of the anti-apoptotic protein BAX is unaltered.
This project consists of two phases carried out in sequence. The first phase will consist of:
- Pharmacological manipulation of organotypic cultures by addition to culture medium of Aβ peptide
fragments;
- Qualitative and quantitative evaluation of the effects the fragments after labeling of live/dead cells by
fluorescent probes (Syto10/Dead Red, Invitrogen, USA or iodidium propide);
- Immunocytochemical analysis of apoptosis with special reference to activation of caspases/PARP1 and
cell cycle pathways (retinoblastoma protein, chk1, cdc2)
- Analysis of relationship between proliferation and apoptosis following visualization of proliferating cells
with immunofluorescence (H3 histone, BrdU) and apoptotic cells with the TUNEL technique. In parallel
cleavage of apoptosis related proteins will be analyzed by Western blotting.
The second phase of the project will consist of:
- Evaluation of the effects of Aβ after biolistic transfection of cultures with the following vectors: ApoAlert
Caspase 3 sensor (Clontech, USA) to monitor the activation of caspase 3 in alive cells; pEYFP-N1 (Clontech,
USA) with insertion of the cDNA for human BCL-2; pHcRed-C1 (Clontech, USA) with insertion of the cDNa
for survivin (Obtained from Dr. Rachel Altura, Ohio, USA);
- Analysis of apoptosis and cell cycle kinetics in transfected cells transfection (as for control cultures
above);
- Analysis of cell calcium dynamics by real time calcium imaging.
Specific experimental procedures
Gene gun technology: We will employ a biolistic transfection procedure (gene gun) to artificially introduce
one or more genes of interest in tissue slices. The procedure involves shooting of slices with gold
microcarriers coated with cDNAs of different constructs encoding for molecule(s) under study and for a
reporter protein (GFP/EYFP/DsRed). Gold microcarriers will be accelerated into tissue by a helium blast
(180 psi) so that they will penetrate cell membranes and reach the cytoplasm of target cells. Transfection
will be performed after 2-5 days in vitro, by a commercial apparatus (Helios Gene Gun System, BioRad,
USA).
Confocal microscopy: Control and transfected cultures will be incubated inside a custom made microscope
stage incubator (Tokai Hit, Japan) and monitored at variable intervals of time under a Leica SP5 laser
confocal microscope. We have already verified that by this set up it will be possible to monitor the cultures
for 4 days or longer under the microscope, and it will be thus possible to study short to medium term
effects on apoptosis. In certain experiments a cell permeant calcium indicator will be used (Oregon Green
Bapta 1AM, Invitrogen, USA)to monitor the intracellular calcium concentration in individual neurons.
Visualization of proliferating cells: a technique based upon incorporation of bromodeoxyuridine (BrdU)
during S phase will be used by adding BrdU directly to culture medium. M and G2 phase cells will be
labeled by immunocytochemistry using a primary antibody directed against phosphorylated H3 histone.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Effects of over-expression of the anti-apoptotic proteins BCL-2 and
survivin on the endoplasmic reticulum stress induced by prion protein
and βamyloid: study of neuroprotection on animal models
MIUR PRIN 2007 (to Laura Lossi)
24 mesi
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Several neurodegenerative diseases are characterized by an initial
endoplasmic reticulum stress followed by the activation of an apoptotic
program leading to cell death and neuronal loss.
The two main goals of this project are:
- analysis of apoptosis induced by prion protein (PrP) e da β-amyloid (Aβ)
in an ex vivo model consisting of murine organotypic cerebellar cultures
obtained from P7-P20 mice. This developmental stage has been proved to
be particularly suitable to our purposes since it is characterized by a
massive apoptotic neuronal death.
- analysis of potentially neuroprotective effects of two anti-apoptotic
proteins (BCL-2 and survivin) onto neuronal death induced by PrP and Aβ.
The project has duration of 24 months, and is organized into two phases,
each corresponding to one of the two main objectives outlined above.
We will employ a multidisciplinary approach consisting of in vitro
culturing techniques, biolistic transfection (gene-gun), confocal
microscopy (medium to long term analysis of transfected cell, reat time
imaging of calcium influx), in situ immunocytochemistry and molecular
biology (immunofluorescence, labeling of proliferating/apoptotic cells),
biochemistry (Western blotting).
All these procedure will eventually aim to obtain a direct correlation
between morphological and functional data.
We expect to obtain a better characterization of the basic biology of PrP
and Aβ in relation to modulation of programmed cell death.
Development of an in vitro system to study the neuroprotective effects
of ghrelin and its derivatives on central neurons
MIUR PRIN 2008
24 mesi
Background and rationale: Ghrelin is a peptide hormone produced by the
endocrine cells of the gastric mucosa. Its most relevant biological action is
the regulation of feeding behavior in the presence of a negative energetic
balance. After discovery at periphery, the hypothalamus has been
identified as the main source of ghrelin in the central nervous system
(CNS). Recent studies show that the hormone has indeed a wide range of
biological effects in central neurons (see Ferrini et al. 2009, Current
Neuropharmacology. 7:37). Among these, we have recently characterized
the role of ghrelin in spinal cord pain neurotransmission (Vergnano et al.
2008. Endocrinology. 149:2306). However, data have been obtained in
support to the notion that ghrelin also plays a role in neuronal
proliferation ,differentiation, protection against apoptosis and
maintenance of a differentiated status (see Chung et al. 2007.
Endocrinology. 148:148). To understand whether or not the effects of
ghrelin in cultured cells are indeed of relevance in vivo it is necessary to
respond to the following question: is the hormone produced in the
stomach capable to cross the blood brain barrier? The importance of this
question appears immediately obvious if one considers that the
physiological levels of ghrelin expression in CNS are relatively low.
General layout and timetable of project: This is a two year project.
During the first year we will first test the antiapoptotic effects of ghrelin
and its derivatives on organotypic cerebellar slices. Then we will develop
an in vitro model based on our previous experience (postnatal cerebellar
slices cultured in vitro under static conditions - see Lossi et al. 2009. Prog
Neurobiol. 88:221) where the integrity of the blood brain barriers is
maintained. This model will be derived according to a recently developed
protocol (Bendfeldt et al. 2007. J Neurosci. 27:3260) where it was
demonstrated that the cultivation of cortical slices in the presence of
FGF2 (basic fibroblast growth factor 2) is ideal to the preservation of the
blood brain barrier. During the second year of the project we will assess
the effects of ghrelin on cell death in relation to integrity of the blood
brain barrier. Aims and methods: The project focuses on the study of the
antiapoptotic effects of ghrelin in a widely established model of
developmental cell death, the post-natal cerebellar cortex. The effects of
the hormone will be compared in the presence/absence of the blood
brain barrier to understand whether or not peripheral ghrelin is capable
to influence the survival of central neurons. To do so we will use a
multidisciplinary approach consisting of in vitro cultures, cell transfection
(gene-gun), confocal microscopy (mid-to-long term analysis of cell
calcium fluxes), immunocytochemistry, molecular biology in situ
(immunofluorescence, labeling of proliferating/dead cells), and
biochemistry (Western blotting). The concurrent employment of all these
procedures will allow obtaining a direct correlation between
morphological and functional data. Project phases: 1. Development of
FGF2 stimulated cerebellar organotypic cultures and assessment of
integrity of the blood brain barrier. 2. Pharmacological manipulation of
cultures with ghrelin, agonists and antagonists. 3. Analysis of cell death by
means of fluorescent probes (Syto 10/Dead Red, Invitrogen, USA). 4.
Immunocytochemical analysis of the molecules involved in the apoptotic
cascade (retinoblastoma protein, cell cycle-related kinases chk1, cdc2), 5.
Analysis of the correlation between apoptosis and cell proliferation (after
labeling with fluorescent probes and/or specific markers). 6. Western
blotting of relevant proteins to study their cleavage in the course of cell
death.
Nome
Contatti
Monica DiLuca
Universita’ degli Studi di Milano
Via Balzaretti, 9
02 50318374
[email protected]
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
CEND and Dept of Pharmacological Sciences
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Synaptic dysfunction in neurodegeneration: characterization of gene
products in functional synaptic networks under physiologicaò and
pathological conditions. This includes characterization of mechanisms
involved in subsynaptic targeting of proteins and protein complexes.
These studies will allow for identificationa nd validation of targets as
molecules responsible for synaptopathies using pharmacological
approaches, siRNA, mimetic peptides and small synthetic molecules. In
addition, these approaches will lead to model
stages of
neurodegenaration in vivo by obtaining innovative animal models for
Alzheimer diseases and other neurodegenerative disorders.
Role of calcium dependent gene expression and protein trafficking in
shaping post synaptic activity: implication for neurodegenerative
diseases.
Cariplo foundation
April 2009 – March 2011
the main goal of the project is to investigate the link between
dyshomeostasis of calcium signalling and early pathogenic events of AD –
both at cellular and molecular level - working on the hypothesis that this
neurodegenerative disorder is a synaptopathy. As an addendum to this,
we do not wish to focus on the role of calcium in neuronal loss but on the
role of initial small changes of calcium homeostasis in synaptic deficits, as
in vivo this correlates better with early neurological disturbances and
cognitive impairment.
Molecular mechanism and behavioural aspects in experimental models of
Parkinson’s disease.
Italian Minister of Health
November 2008 – November 2010
Despite abundant evidences support the role of NMDA receptor complex
in Parkinson’s Disease, only scattered if any information are available at
single subunit level, although this is a much-needed information in order
to identify possible new molecular pathways that can represent a
pharmacological target for the disease as well as strategies to tackle
dyskinesia. Thus, specific aim of the project will be to characterize in
striatal neurons the molecular events responsible for the correct synaptic
localization of NR2A and NR2B subunits of NMDA receptor and to study
the functional consequences of such an event in experimental
parkinsonism and in L-DOPA induced dyskinesia.
“REPLACES” - Restorative Plasticity At Corticostriatal Excitatory Synapses
EU VII Framework Programme
November 2008- October 2012
The main ideas underlying this proposal is that the long-term efficacy of
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
new treatments for Parkinson’s disease (PD) will be conditioned by their
ability to restore, both structurally and functionally, the “synaptic wiring”
of striatal neurons and physiological synaptic plasticity. Accordingly, the
proposal will address the potential restorative effects of either novel
pharmacological treatments or neuronal transplants on the corticostriatal
microcircuitry. Since chronic treatment with the DA precursor L-DOPA
induces in the majority of PD patients a maladapTative plasticity causing
involuntary movements (dyskinesia), innovative strategies should prevent
the development of this disabling condition. The project will characterize
corticostriatal synaptic plasticity from molecular aspects to clinical
neurophysiology involving also behavioural as well as morphological
analysis of the basal ganglia system.
“cPADs” - Cell permeable peptides as drug delivery system: a way
towards innovative therapeutic strategies for neurodegeneration.
EU VII Framework Programme
April 2008- March 2012
The project aims to foster a dynamic pathway between academic
research organizations and private commercial enterprises, based on a
long-term co-operation programme in the field of innovative therapeutic
strategies for neurodegenerative disorders. Through inter-sectoral
industry-academia collaboration, the proposal aims to define appropriate
technologies to target key protein-protein interactions and key
intracellular pathways mediating neurodegeneration and cell death,
addressing both the creation of innovative models and the design of
innovative drugs by means of exploiting the use of Cell-Permeable
Peptides.
Nome
Prof. Renata Bartesaghi
Contatti
Tel +39 051 2091727, +39 051 2091747
Fax +39 051 2091737
e-mail: [email protected]
Istituto/Dipartimento
Dipartimento di Fisiologia Umana e Generale, Università di Bologna
Proposta di ricerca
Study of early onset Alzheimer disease in a model for Down syndrome, the Ts65Dn mouse
Area di interesse identificata:
DEVELOPING COMPETITIVE
ANIMAL MODELS TO STUDY
ALZHEIMER’S DISEASE
Area di interesse identificata:
BASIC RESEARCH
Individuals with Down syndrome (DS) develop neuropathological
features similar to Alzheimer disease (AD) early in life, including
dementia, accumulation of beta-amyloid, and irregular
phosphorylation of tau proteins. Ts65Dn mice have an extra copy of
the distal aspect of mouse chromosome 16, a segment which is
homologous to human chromosome 21 and which contains much of
the genetic material responsible for the DS phenotype. Ts65Dn mice
show developmental delay during the postnatal period as well as
abnormal behaviors in both young and adult animals that may be
analogous to mental retardation. Though the Ts65Dn brain is
normal on gross examination, there is age-related degeneration of
septohippocampal cholinergic neurons and astrocytic hypertrophy,
markers of the AD pathology present in elderly DS individuals.
Moreover, Ts65Dn mice undergo cognitive deterioration beginning
at 6-8 months of age. All this evidence suggests that Ts65Dn mice
may provide a unique model which is suitable for following the
progression of AD.
While AD develops only in some individuals of the general
population, it is the invariable hallmark of DS, implying that trisomic
genes play a paramount role in development of the disease. Indeed,
recent evidence shows that the presence of an extra copy (due to
trisomy 21) of the dual-specificity tyrosine-phosphorylated and
regulated kinase 1A (Dyrk1A) gene results in overexpression of
Dyrk1A and elevated kinase activity in the DS brain. Dyrk1A
phosphorylates tau at several sites, and these sites are
hyperphosphorylated in adult DS brains. Phosphorylation of tau by
Dyrk1A primes its further phosphorylation by glycogen synthase
kinase-3beta (GSK-3beta). These findings strongly suggest a novel
mechanism by which the overexpression of Dyrk1A in DS brains
causes neurofibrillary degeneration via hyperphosphorylating tau.
The putative involvement of Dyrk1A in the etiology of AD implies
that de-regulation of Dyrk1A may be an age-related phenomenon
that ultimately leads to the pathology. In the Ts65Dn mouse brain,
an extra copy of the Dyrk1A gene causes increased expression and
activity of Dyrk1A and results in increased tau phosphorylation.
Consequently, Ts65Dn mice may represent a good model for the
elucidation of the role of Dyrk1A (and other trisomic genes) in the
development of AD.
Area di interesse identificata:
NEW TREATMENT
STRATEGIES
AD is histopathologically characterized by intraneuronal
neurofibrillary degeneration of the abnormally
hyperphosphorylated tau and extracellular β-amyloidosis. The
neurofibrillary degeneration is apparently required for the clinical
expression of AD, and in related tauopathies it leads to dementia in
the absence of amyloid plaques. While normal tau promotes
assembly and stabilizes microtubules, the abnormally
hyperphosphorylated tau disrupts microtubules. Tau is
phosphorylated by a number of protein kinases. Glycogen synthase
kinase-3 (GSK-3) is one of the key kinases required for AD-type
abnormal hyperphosphorylation of tau, which is believed to be a
critical event in neurofibrillary tangle formation. Thus, GSK-3
inhibition represents a very attractive drug target in AD and other
neurodegenerative disorders. The inhibition of abnormal
hyperphosphorylation of tau is one of the most promising
therapeutic targets for the development of disease modifying drugs.
By exploiting the Ts65Dn mouse it will be possible to establish
whether treatments interfering with tau phosphorylation
ameliorate the AD phenotype. More importantly, it will be possible
to establish whether pharmacotherapy during the early life stages
that precede the overt manifestations of AD is able to
prevent/delay the onset of the pathology
Finanziamenti ricevuti
Titolo progetto
PRIN 2008 - Effect of prenatal and early postnatal treatment with
fluoxetine on neurogenesis and dendritic maturation in the Ts65Dn
mouse model for Down's syndrome
Ente finanziatore
MIUR
Durata progetto
2 anni
Abstract del progetto
Cognitive decline in Down mice; pharmacological treatment to
prevent cognitive decline by stimulating neurogenesis
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Sonia Levi
+39 0226434755-53
[email protected]
San Raffaele Scientific Insitute/Division of Neuroscience
Neurodegeneration
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Functional and epidemiological study of neuroferritinopathies:
implications for the role of iron in neurodegenerative disorders.
Telethon
Three years
Iron participates in the oxidative damage common to various
neurodegenerative disorders, such as Alzheimer and Parkinson diseases
and Hallevorden−Spatz syndrome, and iron accumulations have been
frequently observed in the areas of the brain that degenerate. Genetic
abnormalities of some proteins strictly involved in the regulation of iron
metabolism are associated to neurodegeneration. Among them,
neuroferritinopathies are rare autosomal dominant disorders associated
with mutations of the FTL gene that lead to alterations of the C−terminus
of ferritin L−chain. They are characterized by abnormal iron deposition in
the basal ganglia of the brain and by the appearance of ferritin
aggregates. L−ferritin is dedicated to iron storage, and preliminary results
indicate that one of the mutations alters iron availability in cellular
models. The project is aimed at studying the structural and functional
properties of the ferritin mutants, by developing various cellular models,
including primary neurons and glial cells. By this approach, we expect to
provide indications on the molecular mechanisms that are responsible for
both intracellular iron accumulation and protein aggregate formation. In
parallel, we plan to produce knock−in mice models expressing the
mutated ferritins, to study the progress of neurodegeneration as well as
the formation of ferritin aggregates in basal ganglia. Possible therapeutic
approaches, including the use of RNA interference, will be evaluated on
both cellular and animal models. Since DNA variations in FTL gene are not
uncommon, we plan to analyse its mutations, as well as those of other
iron−related genes, in patients with neurological disorders. This study is
aimed to elucidate how iron acts as a cofactor in neurodegenerative
processes.
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Tiziana Antonelli
[email protected]
Dept. of Clinical and Experimental Medicine
Pharmacology Section, University of Ferrara, Italy
Mechanisms of Neurodegeneration : I. Animal models.
Neurochemical and morphological in vivo and in vitro models in
neurodegenerative diseases
BioPharmaNet: Laboratory of research and innovation for the Life
Sciences sector industries (Emilia Romagna, Italy)
2007-2010
In vitro models of neurodegenerative diseases have been developed and
optimized. Cortical cell cultures, mainly containing GABAergic and
glutamatergic neurons, and hippocampal cell cultures, especially
containing glutamatergic neurons, can be used to study the cell
degeneration following neurotoxic insults. The neuronal damage may be
3
3
evaluated by analysing neurochemical ([ H]GABA and [ H]glutamate
uptake and release, biochemical (MTT assay, LDH levels) and
morphological (Hoechst 33258 nuclear staining, MAP-2 immunoreactivity)
parameters. These in vitro experiments, performed in primary neuronal
cultures, cerebral tissue slices and/or synaptosomes, are complementary
to the in vivo microdialysis experiments and by combining them it is
possible to investigate on the efficacy as well as the neurochemical and
molecular mechanisms of new drugs against neurodegenerative
pathologies. In vivo microdialysis technique in unilaterally
6-hydroxydopamine (6-OHDA) lesioned rats allows to understand the
possible relevance of new pharmacological approaches in the treatment
of this pathology. The simultaneous evaluation of the release of several
neurotransmitters such as dopamine, acetylcholine, glutamate and GABA
in the striatum, globus pallidus and/or other close related brain nuclei of
the indirect and direct pathways of the basal ganglia, gives the
opportunity to evaluate the efficacy of new therapeutic and genetic
strategies in the control of the “motor” pathways both in physiological
and pathological conditions.
2. PHARMACOLOGY
A- Drug Design
Nome
Contatti
Istituto/Dipartimento
Nino Russo
[email protected]
Dipartimento di Chimica, Università della Calabria, via P. Bucci, cubo14c,
87036 Rende (CS), Italy
Proposta di ricerca
Alzheimer_s disease (AD) is characterized by the deposit of extracellular amyloid plaques, which consist
predominantly of the aggregated peptide amyloid-b (Ab). The aggregation mechanism is not very well
understood, but it is proposed that aggregated forms of Ab are neurotoxic because of the
production of reactive oxygen species (ROS). Copper and other metal ions (e.g. Zn) have been found to
accumulate in amyloid plaques. Moreover, Cu ions have been proposed to be linked to the aggregation of
the Ab peptide and/or to the increased generation of ROS. In this context, Cu II coordination to Ab
becomes a key feature, because it will directly affect the structure of the peptide and, hence, the
aggregation behaviour.
The catalytic ROS production of the Cu II ions coordinated by Ab will also be governed by the binding
mode. Therefore, this system has been extensively studied. However, the unambiguous identification of
the Cu II ligands has remained difficult and no real consensus has emerged in the literature yet. It
has been shown that all of the ligands of the high affinity Cu II site are contained in the Ab16 N-terminal
domain (DAEFRHDSGYEVHHQK); hence, this soluble fragment has been widely used as a model for the CuII
binding site in the native Ab40/42 peptides. Although a lot of experimental studies performed with a wide
range of methods, the analysis of the results difficult and impaired the unambiguous identification of the
Cu II ligands.
Modern theoretical and computational methods can give a significant contribution to the knowledge of
the elementary mechanisms at atomistic level on a series of open questions such as: i. unambiguous
identification of the metal coordination site; ii. the role of the metal coordination in the conformational
change of the metallated protein; iii. The effect of the environment on the complex formation; iv. Design
of new and specific drugs for the metal chelation and sequestration.
For the development of this research program, sophisticated computational protocols as well as
theoretical methods will be implemented and used.
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Basic research
Development of computational and theoretical methods based on the
Density Functional theory and their application on systems with biological
significance.
Università della Calabria
Due anni
New algorithms and computational protocols based on the density
functional theory should be proposed and tested. The applications should
be essentially devoted to the study of complex biological systems
Nome
Carlo Melchiorre
Contatti
e-mail: [email protected] ph: 051-2099706
Istituto/Dipartimento
Dipartimento di Scienze Farmaceutiche
Università di Bologna
Via Belmeloro, 6, 40126 Bologna
Proposta di ricerca
Innovative drug discovery strategies for Alzheimer’s disease
Area di interesse identificata
New treatment strategies
Alzheimer’s disease (AD) is a major neurodegenerative disease
affecting a substantial proportion of elderly population and
imposing a significant burden on public health. For AD there is still
no cure, and only few symptomatic treatment options are available
to the 30 million patients worldwide. Driven by the clear unmet
medical need and a better understanding of the AD biology and
pathophysiology, several lead candidates have progressed to preclinical testing in the last decade. However, to date none has been
successful in late-stages clinical trials. Currently, AD is best
characterized as a multidysfunctional condition, where interrelated
molecular events result in amyloid formation, tau abnormalities,
amyloid accumulations, and loss of acetylcholine neurotransmission. Currently available drugs and most of the drugs under
development target only one of these mechanisms. This may
explain why they exhibit limited effectiveness. Therefore,
therapeutic tools able to confront this complexity are urgently
needed. Another issue to take into account in AD drug discovery is
that AD patients are susceptible to a wide range of concomitant
medical conditions (co-morbidity), including hypertension, vascular
diseases, diabetes, which can often be associated. There is
therefore a need for therapeutic tools to be tailored to their specific
conditions.
In this respect, we have recently proposed that multitarget ligands,
i.e. single chemical entities able to modulate simultaneously
multiple targets, might represent the best pharmacological option
for both tackling the multifactorial nature of AD and simplifying AD
patient therapeutic regimen. Furthermore, from a chemical biology
perspective, we envisaged that rationally designed multitarget
chemical probes have promise for studying the complex
mechanisms underlying neurodegeneration.
Finanziamenti ricevuti
Titolo progetto
Principal Investigator: “Neuroreceptors machinery: design and
synthesis of molecules useful for their characterization and
modulation and for the treatment of related dysfunctions”
(20073EWPF9_003;)
Ente finanziatore
Ministero dell’Istruzione, dell’Università e della Ricerca (€ 490,000)
Durata progetto
22/09/2008-21/09/2010
Abstract del progetto
The present project regards the design, synthesis and preliminary
pharmacological evaluation of ligands and modulators of
neurotransmitter receptor systems. The knowledge of structure and
functions of neurotransmitter receptor systems is critical to design
and develop drugs useful to treat the numerous pathologies that
depend on central and peripheral nervous transmission
dysfunctions. As a matter of fact, it is well known that severe
pathologies such as pain, anxiety, mental conditions, and
neurodegeneration are due to receptor dysfunctions. Some of these
pathologies have a tremendous social impact as they involve large
population segments, in particular aged people.
Titolo progetto
Principal Investigator: “Development of Innovative Methodologies
for the Identification and Synthesis of New Bioactive Molecules:
application to the Alzheimer's disease” (FIRB 2003 RBNE03FH5Y;) €
Ente finanziatore
Ministero dell’Istruzione, dell’Università e della Ricerca (€
2,871,000)
Durata progetto
12/09/2005-11/09/2008
Abstract del progetto
The modern Medicinal Chemistry is nowadays requiring an actual
multidisciplinary approach and the development of new
technologies either to accelerate the whole discovery process, or to
provide better and safer candidate drug. In this context, the goals of
the present project are both to develop innovative technological
frameworks for molecular recognition and combinatorial chemistry,
and to validate the applicability of such techniques through the
production of new therapeutics in the field of the Alzheimer's
disease (AD)
Nome
Contatti
Istituto/Dipartimento
Prof. Federico Da Settimo, Prof. Claudia Martini
[email protected], [email protected]
Department of Pharmaceutical Science, Department of Psychiatry,
Neurobiology, Pharmacology, and Biotechnology - University of Pisa
Proposta di ricerca
Development of labelled ligands specifically targeting the Translocator Protein 18 kDa in Alzheimer
Disease.
The Translocator Protein (TSPO) is a mitochondrial transmembrane protein involved in a variety of
biological processes, such as steroidogenesis, apoptosis induction and inflammation, and it is differently
expressed under physio/pathological conditions, in brain cells (astrocytes and microglia). A dramatic
increase in TSPO levels occurs in glial cells in response to brain injury and inflammation, so that TSPO can
be considered as a biomarker of ongoing brain insults. The activation of microglial cells and the
consequent up-regulation of the TSPO sites have been demonstrated in both normal ageing and in brains
of patients affected by Alzheimer disease (AD), stroke and multiple sclerosis. Increased TSPO expression
has been observed In this view, the use of TSPO ligands could efficiently reveal microglia activation, so
representing a useful tool in brain imaging, both in vivo and in post-mortem analysis.
Our group has gained experience in the synthesis of heterocyclic compounds acting at the
mitochondrial translocator protein (TSPO). As well documented by several international publications, over
the course of several years the research group has also gained experience in the evaluation of the
biological activity of new synthesised compounds, through high yield methodologies (Primofiore, G., et al.
J. Med. Chem. 2004, 47, 1852; Chelli, B., et al Chembiochem. 2005, 6, 1082; Taliani, S., et al. J. Med. Chem.
2007, 50, 404; Da Settimo, F., et al. J. Med. Chem. 2008, 51, 5798; Chelli, B., et al. J. Cell. Biochem. 2008,
105, 712.)
The gained experience have generated significant efforts of the group to apply TSPO ligands as
marker of “active” brain pathology, in line with multiple research groups throughout the world.
In the present project we aim to design, synthesise and evaluate novel specific TSPO probes
featuring the indolglyoxylamide scaffold, suitable as useful tools in brain imaging. A number of
radiolabelled ligands targeting TSPO will be developed as powerful tools to image and measure the
expression levels of the protein in humans, opening up the opportunity for the discovery of specific PET
ligands for TSPO. PET studies of TSPO could offer quantitative follow-up of neuroinflammation, an
advantage for biomarker-type measurement during drug development and early clinical development.
Preliminarily, studies are in progress with monkeys.
Moreover, fluorescent ligands, that represent a safer, faster, and less expensive alternative to
radioligands, will be developed for usage in biomedical research. In particular we will investigate a series
of fluorescent probes characterized by the presence of a chemoreactive group able to bind the receptor
protein irreversibly and covalently, in order to overcome the fact that tissue experimental procedures
using a labelled ligand often cause the alteration of the chemical equilibrium and the subsequent loss of
the bound fluorescent ligand. The presence of these two devices (a chemoreactive group and a fluorescent
chromophore) on a single molecule may offer a multiplicity of advantages, both in protein
purification/characterization, and in protein cell visualization/density determination in both cell cultures
and tissue slices derived from post-mortem brains.
Pursuing of these goals will be realized in strict collaborations with the research groups of Prof.
Silvia Selleri (University of Florence).
Area di interesse identificata
Finanziamenti ricevuti
Drug development for AD Diagnosis, Monitoring and Brain imaging
Titolo progetto
New fluorescent tools targeting the translocator protein (TSPO) as
diagnostics for cancer and neurodegenerative diseases.
Ente finanziatore
Durata progetto
University of Pisa
24 months
Abstract del progetto
The proposed research is addressed to develop innovative fluorescent
probes targeting the 18 kDa Translocator Protein (TSPO) (previously
known as the peripheral-type benzodiazepine receptor) as promising
molecular imaging (MI) tools to be widely applied in studying TSPO
location, turn-over and interaction as well as in diagnosing TSPO related
pathologies. TSPO is a protein mainly expressed in mitochondria, which is
involved in cell proliferation, apoptosis, immunomodulation and
steroidogenesis. TSPO basal density is altered in pathologies in which a
perturbation of cell survival/death processes has been evidenced such as
cancer and neurodegenerative disorders and alterations in TSPO
subcellular distribution have been correlated with the malignancy grade
of cancer cells. Therefore, TSPO has been suggested as a promising drug
target as well as a valid diagnostic marker of state and progression of
such pathologies and has become an interesting target in MI. Currently,
MI has emerged as an important multidisciplinary area, which involves
radiology, biology, biochemistry, physics, engineering, medicine and
synthetic chemistry, essential in providing potent imaging agents. Based
on our previous experiences, the aim of this research program is to
design, synthesize and biologically characterize/validate new reversible
and irreversible fluorescent 2-phenylindol-3-ylglyoxylamide ligands with
high affinity and selectivity for TSPO. The development of these
innovative fluorescent TSPO ligands will offer a wide range of advantages
for their applications in molecular basic studies as well as in diagnosis.
Nome
Alberto Cassetta
Contatti
[email protected] - Tel: ++39 040 226881
Istituto/Dipartimento
CNR – Istituto di Cristallografia UOS Trieste (Dip. Progettazione
Molecolare)
Proposta di ricerca
Structural studies of acetylcholinesterases and their inhibitors: implications for the design of new anti-Alzheimer
drugs
Acetylcholine esterase (AChE) plays a crucial role in the regulation of signal transmission, by terminating the action of
acetylcholine at the cholinergic synapse and muscular junctions. AChE inhibitors are used in the symptomatic
treatment of pathologies such as Alzheimer's disease. The 3D crystal structures of a number of AChE complexes with
reversible and irreversible inhibitors have been determined. The structural information accompanied by kinetic
studies will allow a detailed understanding of the molecular determinants underlying the mechanism of action of this
enzyme. Furthermore these results are expected to offer a rational framework for the design and synthesis of
compounds with optimal pharmacokinetic and pharmacodynamic properties.
Structural studies of neurotrophins (NGF), proneurotrophins(proNGF) and their interactions with their receptors
TrkA, p75, sortilin) and a panel of neutralizing antibodies(MNAC13, αD11)
Neurotrophins such as nerve growth factor (NGF) are involved in the development and maintenance of neurons both
in the central and peripheral nervous system, and have been implicated as potential therapeutic target molecules in
the Alzheimer's disease. NGF exerts its biological function by binding to TrkA receptor promoting dimerization,
catalytic auto-phosphorylation of intracellular tyrosine residues and triggering of the signal transduction cascade.
Until recently, no clear biological role had been attributed to the proneurothrophin (proNGF) peptide, except helping
in folding and secretion of mature NGF. Moreover, proNGF was found to be the predominant form of NGF in brain
and to be increased in Alzheimer's disease and to induce p75NTR dependent apoptosis in cultured neurons. The
specific receptor for proNGF is sortilin. We are investigating the structural and functional bases of the biological
differences between NGF and proNGF. The monoclonal antibodies MNAC13 and αD11 are potent antagonists that
prevent the NGF-TrkA interactions in vivo systems. In order to gain deeper insight into the molecular basis of the
observed high affinity of NGF for TrkA, we are investigating the 3D structures of both neutralizing antibodies and of
the respective complexes. Detailed structural information on their antigenic recognition is expected to aid in the
development of analogs as antagonists or agonists of neurotrophins that may have greater affinity or specificity for
further experimental and therapeutic applications.
Structural studies of Hydroxysteroid Dehydrogenases and their interactions with phytoestrogens offlavonoid
nature.
17β-Hydroxysteroid dehydrogenases (17β-HSDs) are enzymes which play a crucial role in steroid hormones action by
acting on the final steps of estrogen or androgen biosynthesis. Given their ability to interconvert a hormonally
inactive (or less active) steroid to its active form, these enzymes operate a pre-receptorial regulation of steroid
action. Several distinct 17β-HSDs isozymes can be found in human, principally distinct by their tissue type and cellular
localization, homology sequence and metabolic pathway. In humans different 17β-HSDs have been linked to several
pathological forms: prostate and breast cancer (17β-HSD type 1) and Alzheimer’s disease (17β-HSD type 10) among
others. We have recently undertaken a structural study on a fungal 17β-HSD which share a remarkable structural
similarity with both type 1 and 10 of human 17β-HSD. Our studies were aimed to investigate the use of the fungal
enzyme as possible model for the understanding at the molecular level the interaction mechanism of 17β-HSDs with
specific and less-specific inhibitors. In particular we are investigating the inhibitory action of phytoestrogens,
principally of flavonoid nature, towards 17β-HSD type 1, which have been demonstrated to be inactivated at the
μmol level by certain flavonoids and coumestans. Moreover, given the lack of biochemical and structural data about
the effects of flavonoids on type 10 17β-HSD, we intend to further investigate this subject by a
biochemical/biophysical approach combined with crystallographic methods by studying the inhibition properties of
the above compounds toward the human 17β-HSD homologue. Because both mutations of the gene encoding this
enzyme and its overexpression give raise to a distinct type of mental retardation (17--β-Hydroxysteroid
Dehydrogenase X Deficiency; OMIM #300438) and it has been shown to be a molecular link between mitochondrial
dysfunction and Alzheimer’s disease by direct binding of the β-amyloid peptide, our approach will provide a valuable
understanding of the role played by this enzyme in the occurrence of these neuropathies.
LUSTBADER J, CIRILLI M., LIN C., XU H.W., CASPERSEN C., WANG N., TAKUMA K., CHEN X., CHANEY M. TRINCHESE
F., LIU S., KUPPUSAMY P., ZEWIER Z., ARANCIO O., STERN D., YAN S.D., WU H.: “ABAD Directly Links Aβ to
Mitochondrial Toxicity in Alzheimer’s Disease”. Science (2004) 304, 448-52
Paoletti F, Covaceuszach S, Konarev PV, Gonfloni S, Malerba F, Schwarz, E,Svergun DI, Cattaneo A, Lamba D. “Intrinsic
structural disorder of mouse proNGF”. Proteins. 2009 Jun;75(4):990-1009.
Covaceuszach S, Cassetta A, Konarev PV, Gonfloni S, Rudolph R, Svergun DI, Lamba D, Cattaneo A. “Dissecting NGF
interactions with TrkA and p75 receptors by structural and functional studies of an anti-NGF neutralizing antibody”.
J Mol Biol. 2008 Sep 12;381(4):881-96.
Bartolucci C, Haller LA, Jordis U, Fels G, Lamba D. “Probing Torpedo californica acetylcholinesterase catalytic gorge
with two novel bis-functional galanthamine derivatives”. J Med Chem. 2010 Jan 28;53(2):745-51.
Cattaneo, S. Covaceuszach, D. Lamba: “Method for the humanization of antibodies and
humanized
antibodies thereby obtained”. Patent WO/2005/061540
Area di interesse
identificata
Structural Neurobiology
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Not Applicable
2. PHARMACOLOGY
B-Drug screening
Nome
Laura Calzà
Contatti
[email protected]
Tel. +39 051 2097947
Cell. +39 335 310979
Istituto/Dipartimento
DIMORFIPA
Proposta di ricerca
Area di interesse identificata
Development of competitive
animal models for AD
GLP grade animal facility and behavioural testing for cognitive
performance in AD animal models using video tracking and
computer analysis; screening of new molecules after acute
administration
Basic research
Brain metabolism of thyroid hormone: a risk factor for AD? We are
interested in exploring possible connections between thyroid
hormone content, receptor and activating enzyme expression, APP
metabolism and Ab toxicity in the brain in normal and AD mice.
Recent advances in knowledge on thyroid hormone brain
metabolism and molecular biology of nuclear and membrane
thyroid hormone receptors and transporters offer new possible
targets to explore this topic.
New treatment strategies
Nome
Contatti
Iliana Ferrero Fortunati
[email protected]
Phone: +39-0521905600
Fax: +39-0521905604
Mobile : +39-3480030063
Istituto/Dipartimento
Dipartimento di Genetica, Biologia dei microrganismi, Antropologia,
Evoluzione – Università degli Studi di Parma
Parco Area delle Scienze 11/A - Campus
43100 Parma
Proposta di ricerca
“Yeast model to seek for molecules which can reduce mitochondrial mutability”
Mitochondrial dysfunction is a prominent feature of Alzheimer’s Disease (AD) and there is growing body
of evidence that mitochondrial dysfunction plays a crucial role in the pathogenesis or influences the risk of
AD.
The Yeast Genetics Group of University of Parma, coordinated by of Prof. Iliana Ferrero, has a long lasting
experience in the use of Saccharomyces cerevisiae as a model system to study mitochondrial mutability to
address the question of whether there is a molecular relationship between the pathology and the specific
mutation and to study the pathogenic mechanisms of human disease associated to mitochondrial DNA
mutations. The yeast group has a forty years experience on yeast genetics and a ten years experience in
the use of yeast model of mitochondrial diseases (in collaboration with Dr. Massimo Zeviani, Unit of
Molecular Neurogenetics, IRCCS Foundation Neurological Institute FC. Besta, and with Prof. Giacomo
Comi, Department of Neurological Sciences and Centre of Excellence on Neurodegenerative Diseases,
University of Milan). We have created yeast models of several mitochondrial pathologies due to mutations
in a relevant number of genes involved in mitochondrial DNA stability. Moreover, we have already used
the S. cerevisiae model system to study the effect of molecules on mitochondrial DNA mutability and we
have identified molecules able to reduce mitochondrial mutability.
Specific aims
The main scope of our proposal is the use of S. cerevisiae models already existing in our collection and,
when necessary, create new yeast models to test a pharmacological approach to therapy. To this aim we
propose to subject yeast model to chemicals (chemical library) screening to prospect new therapeutic
drugs able to reduce either extended or point mutability or both.
Publications of Iliana Ferrero on yeast model of mitochondrial disease
Dallabona, C., Marsano, R.M., Arzuffi, P., Ghezzi, D., Mancini, P., Zeviani, M., Ferrero, I., Donnini, C. (2010).
Sym1, the yeast ortholog of the MPV17 human disease protein, is a stress-induced bioenergetic and
morphogenetic mitochondrial modulator. Hum Mol Genet 2010 Jan 15. [Epub ahead of print]
Ghezzi D, Goffrini P, Uziel, G, Horvath R, Klopstock T, Lochmaller H, D'Adamo P, Gasparini P, Strom TM,
Prokisch H, Invernizzi F, Ferrero I, Zeviani M (2009). SDHAF1, encoding a LYR complex-II specific assembly
factor, is mutated in SDH-defective infantile leukoencephalopathy. Nature Genetics, 41, 654-656.
Di Fonzo, A., Ronchi, D., Lodi, T., Fassone, E., Tigano, M., Lamperti, C., Corti, S., Bordoni, A., Fortunato, F.,
Nizzardo, M., Napoli, L., Donadoni, C., Salani, S., Saladino, F., Moggio, M., Bresolin, N., Ferrero, I., Comi,
G.P. (2009). The mitochondrial disulfide relay system protein GFER is mutated in autosomal-recessive
myopathy with cataract and combined respiratory-chain deficiency. Am J Hum Genet 84 594-604.
Ghezzi, D., Goffrini, P., Uziel, G., Horvath, R., Klopstock, T., Lochmaller, H., D'Adamo, P., Gasparini, P.,
Strom, T.M., Prokisch, H., Invernizzi, F., Ferrero, I., Zeviani, M. (2009). SDHAF1, encoding a LYR complex-II
specific assembly factor, is mutated in SDH-defective infantile leukoencephalopathy. Nature Genetics,
41, 654-656
Goffrini, P., Ercolino, T., Panizza, E., Giachè, V., Cavone, L., Chiarugi, A., Dima, V., Ferrero, I., Mannelli, M.
(2009). Functional study in a yeast model of a novel succinate-dehydrogenase subunit B gene germline
missense mutation (C191Y) diagnosed in a patient affected by a glomus tumor. Hum Mol Genet 18,
1860-1868.
Spinazzola, A., Invernizzi, F., Carrara, F., Lamantea, E., Donati, A., Dirocco, M., Giordano, I., MeznaricPetrusa, M., Baruffini, E., Ferrero, I., Zeviani, M. (2009). Clinical and molecular features of mitochondrial
DNA depletion syndromes. J Inherit Metab Dis 32 143-158.
Galassi, G., Lamantea, E., Invernizzi, F., Tavani, F., Pisano, I., Ferrero, I., Palmieri, L., Zeviani, M. (2008).
Additive effects of POLG1 and ANT1 mutations in a complex encephalomyopathy. Neuromuscul Disord
18 465-70.
Massa V, Fernandez-Vizarra E, Alshahwan S, Bakhsh E, Goffrini P, Ferrero I, Mereghetti P, D'Adamo P,
Gasparini P, Zeviani M. (2008). Severe Infantile Encephalomyopathy Caused by a Mutation in COX6B1, a
Nucleus-Encoded Subunit of Cytochrome C Oxidase. Am J Hum Genet. 82 1281-1289.
Massa, V., Fernandez-Vizarra, E., Alshahwan, S., Bakhsh, E., Goffrini, P., Ferrero, I., Mereghetti, P.,
D'Adamo, P., Gasparini, P., Zeviani, M. (2008). Severe infantile encephalomyopathy caused by a
mutation in COX6B1, a nucleus-encoded subunit of cytochrome c oxidase. Am J Hum Genet 82 12811289.
Barberio, C., Bianchi, L., Pinzauti, F., Lodi, T., Ferrero, I., Polsinelli, M., Casalone, E. (2007). Induction and
characterization of morphologic mutants in a natural Saccharomyces cerevisiae strain. Can J Microbiol 53
223-230.
Baruffini, E., Ferrero, I., Foury, F. (2007). Mitochondrial DNA defects in Saccharomyces cerevisiae caused
by functional interactions between DNA polymerase gamma mutations associated with disease in
human. Biochim Biophys Acta 1772 1225-35.
Fernandez-Vizarra E, Bugiani M, Goffrini P, Carrara F, Farina L, Procopio E, Donati A, Uziel G, Ferrero I,
Zeviani M. (2007). Impaired complex III assembly associated with BCS1L gene mutations in isolated
mitochondrial encephalopathy. Hum Mol Genet. 16 1241-1252.
Valente, L., Tiranti, V., Marsano, R.M., Malfatti, E., Fernandez-Vizarra, E., Donnini, C., Mereghetti, P., De
Gioia, L., Burlina, A., Castellan, C., Comi, G.P., Savasta, S., Ferrero, I., Zeviani, M. (2007). Infantile
encephalopathy and defective mitochondrial DNA translation in patients with mutations of
mitochondrial elongation factors EFG1 and EFTu. Am J Hum Genet 80 44-58.
Baruffini, E., Lodi, T., Dallabona, C., Puglisi, A., Zeviani, M., Ferrero, I. (2006). Genetic and chemical rescue
of the Saccharomyces cerevisiae phenotype induced by mitochondrial DNA polymerase mutations
associated with progressive external ophthalmoplegia in humans. Hum Mol Genet 15 2846-55.
Spinazzola, A., Viscomi, C., Fernandez-Vizarra, E., Carrara, F., D'Adamo, P., Calvo, S., Marsano, R.M.,
Donnini, C., Weiher, H., Strisciuglio, P., Parini, R., Sarzi, E., Chan, A., DiMauro, S., Rötig, A., Gasparini, P.,
Ferrero, I., Mootha, V.K., Tiranti, V., Zeviani, M. (2006). MPV17 encodes an inner mitochondrial
membrane protein and is mutated in infantile hepatic mitochondrial DNA depletion. Nat Genet 38 570-5.
Fontanesi, F., Viola, A.M., Ferrero, I. (2006). Heterologous complementation of the Klaac null mutation of
Kluyveromyces lactis by the Saccharomyces cerevisiae AAC3 gene encoding the ADP/ATP carrier. FEMS
Yeast Res 6 414-20.
Lodi, T., Bove, C., Fontanesi, F., Viola, A.M., Ferrero, I. (2006). Mutation D104G in ANT1 gene:
Complementation study in Saccharomyces cerevisiae as a model system. Biochem Biophys Res Commun
341 810-5
Palmieri L, Alberio S, Pisano I, Lodi T, Meznaric-Petrusa M, Zidar J, Santoro A, Scarcia P, Fontanesi F,
Lamantea E, Ferrero I, Zeviani M (2005). Complete loss-of-function of the heart/muscle-specific adenine
nucleotide translocator is associated with mitochondrial myopathy and cardiomyopathy. Hum Mol Genet
14 3079-88.
Fontanesi F, Palmieri L, Scarcia P, Lodi T, Donnini C, Limongelli A, Tiranti V, Zeviani M, Ferrero I, Viola AM
(2004). Mutations in AAC2, equivalent to human adPEO-associated ANT1 mutations, lead to defective
oxidative phosphorylation in Saccharomyces cerevisiae and affect mitochondrial DNA stability. Hum Mol
Genet 13 923-34.
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Yeast model for human health: Neurodegenerative diseases;
Alzheimer’s disease
“Identification and characterization of nuclear genes responsible for human
mitochondrial disorders” Project coordinated by Massimo Zeviani PhD, Unit of
Molecular Neurogenetics, IRCCS Foundation Neurological Institute FC. Besta.
Telethon Italy – Tree years project – 2008-2010
Three years
Albeit still insufficiently acknowledged, mitochondrial disease encompasses a
substantial fraction of human inherited disorders. The inclusive term
"mitochondrial medicine" has recently been coined to account for the broad
functional implications of mitochondria in normal and disease conditions.
Mitochondrial disorders are very heterogeneous, can affect any organ, at any
age, and responsible genes are still missing in most cases. These features make
the diagnosis of mitochondrial disease difficult, and in many instances cause
these disorders to be overlooked. Effective therapy is missing as well.
Nevertheless, great progress has been made in this field in the past decade,
thanks to the discovery of new genes relevant to disease, and the investigation
of the physiopathology of these disorders through a multidisciplinary approach.
In recent years we have been finding several new disease genes, which are
responsible for well−defined mitochondrial syndromes. However, the function of
these genes is still completely or largely unknown.
We shall also create disease models user the friendly systems yeast recombinant
strains. The study of yeast model will help us understand the physiopathology of
the disorders caused by the defective genes, gain insight on their function and
related metabolic pathways, and devise sensible and specific therapeutic
strategies.
Nome
Luca Ferraro
Contatti
[email protected]
Istituto/Dipartimento
Dept. of Clinical and Experimental Medicine
Pharmacology Section, University of Ferrara, Italy
Proposta di ricerca
Area di interesse
identificata
Pharmacology: B. Drug screening .
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Innovative technologies for the therapy of the neurodegenerative
diseases
NeuroBioTech: Innovative technologies for the therapy of the
neurodegenerative diseases (Emilia Romagna, Italy)
2009-2012
The aim of NeuroBioTech is the development of a precompetitive
research activity to find new therapeutic strategies for the treatment
of Alzheimer disease (AD) and other correlated neurodegenerative
pathologies. The experimental activity of our research unit is aimed
to develop and optimize in vivo models (microdialysis, behavioural
tests) and ex vivo models (cerebral tissue), which will be applied in
preclinical testing activity for the screening of molecules with antiAlzheimer activity and for the identification of new therapeutic
targets in AD. The eventual alterations of
endogenous
neurotransmitter levels (glutamate, GABA, noradrenaline,
dopamine) will be evaluated in the above reported in vivo and ex
vivo experimental models. In particular, the eventual modifications
of neurochemical signal will be analysed in mice with mutations of
APP and/or PS and/or tau. Furthermore, it will be studied if the
CHF5074 (new modulator of gamma secretases) administration will
be able to correct these neurochemical alterations. Then, the effect
of CHF5074 on the APP metabolism and on APP molecular
interactions will be studied.
2. PHARMACOLOGY
C- Drug Delivery
Country: Italy
Contact person: Prof. MA Vandelli, Dr. G Tosi ([email protected])
Università di Modena e Reggio
Date: 14-01-2009
I)
Strategic Issues (Justification of the importance of the issue)
NANOTECHNOLOGY FOR HEALTH: THE APPLICATION IN THE BRAIN DISEASES
The application of nanotechnology to health raises high expectations for a more efficient and affordable
healthcare and has the potential of delivering promising solutions to many illnesses. Even if several areas of
medical care could benefit from the advantages that nanotechnology can offer, a selective CNS drug
delivery and targeting could improve the therapy of brain diseases which have a tremendous negative
impact not only on the patient himself but also on the whole society and linked social and insurance
systems.
The project could be also the answer to the fragmentation in nanomedicine research in Europe establishing
a clear strategic vision in the area.
In order for CNS drugs to penetrate to the brain tissue, they must pass through the blood brain barrier
(BBB). Many of the compounds that otherwise would be effective in treating CNS diseases are excluded
from reaching a sufficient concentration in the brain tissue and producing the desired therapeutic effect. It
has been estimated that only 2% of the possible CNS therapeutic compounds can cross the BBB.
Thus, drug delivery to the brain is a challenge, because this tissue benefits from a very efficient protective
barrier. Even if several disorders and diseases that affect the brain can lead to some loss of BBB integrity,
this barrier represents an insurmountable obstacle for many drugs able to treat pathologies in which the
BBB integrity is preserved.
The same mechanisms that protect the brain from foreign substances also restrict the entry of many
potentially therapeutic agents. The blood brain barrier is the major barrier to the passage of active
molecules from the blood compartment to the brain. It is located at the level of the brain capillaries, where
there is a convergence of different cell types: endothelial cells, pericytes, astrocytes and microglias. The
brain microvessel endothelial cells that form the BBB, display important morphological characteristics such
as the presence of tight junctions between the cells, the absence of fenestrations and a diminished
pinocytic activity, that together help to restrict the passage of compounds from the blood into the
extracellular environment of the brain.
A promising strategy for the delivery of therapeutics to CNS could be to associate drugs without any
modification (prodrugs, for example) to colloidal carriers. These vehicles could deliver numerous drug
molecules at specific sites by coupling ligands to the surface of the colloids which can be administered
intravenously for chronic treatment. Moreover, owing to their poor stability in biological fluids, rapid
enzymatic degradation, unfavourable pharmacokinetic properties and lack of diffusion towards the CNS,
biotech drugs (peptides, proteins, genes and antisense drugs) may be advantageously formulated in brain
targeted protective nanocontainers.
Colloidal drug carriers include micelles, emulsions, liposomes and nanoparticles (nanospheres and
nanocapsules). So far, only liposomes and nanoparticles have been largely exploited for brain drug delivery.
Recently, the first liposomal preparation of a drug reached the market: it is a formulation of the antitumor
drug Doxorubicin loaded into PEGyilated liposomes for the i.v. treatment of brain tumors (commercial
formulation Caelyx). In contrast to liposomes, and despite the abudance of experimental works and
achievements in the field of polymeric nanoparticle (Np) technology that show the ability of properly
functionalized nanoparticles to cross the BBB, no nanoparticle-based drug formulations have been
marketed so far. Drugs encapsulated into poly(butylcyanoacrylate) (PBCA) Np covered by the surface active
agent polysorbate 80 (Doxorubicin. Loperamide, Dalargin, the anticonvulsivant MRZ 2/576), on the surface
of HSA Np (the model drug loperamide), unable to cross BBB when administered alone, were able to exert
an appropriate pharmacological effect. Despite interesting results, PBCA Np have limitations that may
preclude or at least limit their potential clinical applications; its use for humans is at the moment not
approved by FDA. Thus there is a need for the development of nanoparticles based for example on the
polymer poly(D,L-lactide-co-glycolide (PLGA), which is approved by FDA for human use, and very promising
results were found in the literature.
So far, only a low amount of injected nanoparticle dose is able to reach the brain (in the order of 1%); thus
active targeting of the BBB represents a promising non invasive strategy for improving drug delivery to
brain. In fact, the use of ligands specific for receptors present in high percentage on the BBB could
represent the way to realize a selective delivery of nanoparticles, and thus of the embedded drugs, to CNS.
With this aim, positive results were obtained when nanocarriers have been conjugated to ligands (OX26
mAb; transferrin, apolipoproteins or surface-active agents (polysorbate-80) able to bind some of the
apolipoproteins present in blood that confer the ability to cross the BBB, insulin, folic acid) for active
targeting, but other ligands have to be discovered and tested for their ability to carry into CNS drug-loaded
nanoparticles.
The application of nanotechnology to health raises high expectations for a more efficient and affordable
healthcare and has the potential of delivering promising solutions to many illnesses. Even if several areas of
medical care could benefit from the advantages that nanotechnology can offer, a selective CNS drug
delivery and targeting could improve the therapy of brain diseases which have a tremendous negative
impact not only on the patient himself but also on the whole society and linked social and insurance
systems.
The project could be also the answer to the fragmentation in nanomedicine research in Europe establishing
a clear strategic vision in the area.
II)
Priority areas
Health; Neurodegenerative Diseases
Innovative strategies for the treatment of brain pathologies
Nanotechnology applied to Health, Nanomedicine
III)
Impact
The pharmaceutical treatment of central nervous system (CNS) disorders is the second largest area of
therapy, following cardiovascular disease. U.S. sales for CNS drugs exceeded $ 53 billion in 2002 to 2003.
CNS disorders are five of the top 10 causes of disability. Stroke is the third leading cause of death and costs
the economy $ 40 billion annually. Fifteen million people suffer from Alzheimer’s disease, which is the
second most expensive disease to the economy at $ 100 billion annually. Many brain diseases do not have
satisfactory treatments. Clearly, CNS disorders are an important current and future priority for the
pharmaceutical industry.
Moreover, the development of new drug molecules is expensive and time-consuming. The average cost and
time for the development of a new chemical entity are much higher (approximately $ 500 million and 10-12
years, respectively) than those required to develop a novel drug delivery system. Hence, there is a need for
evolving an existing drug molecule from a conventional form to a novel delivery system that can
significantly improve its performance in terms of patient compliance, safety and efficacy by targeting to the
specific site.
Partner involved:
•
Italy (ACADEMY) (PROPONENT) (Prof. Vandelli, Department of Pharmaceutical Sciences, Dean of
Faculty of Pharmacy, University of Modena and Reggio Emilia, T.e.Far.T.I.)
o Nanotechnology, nanoparticles, liposome, surface modification, derivatization of biodegradable
polymers, in vivo experience on animals
o In vivo pharmacology expertise on CNS drug efficacy
o Preparation and loading of engineered Np with selected drugs or active substance
•
Switzerland (ACADEMY)
o Nanotechnology, Coupling methodology with preformed Np , in vitro tests on BBB culture
•
England (ACADEMY)
o Nanomedicines based on novel nanomaterials of synthetic and biological nature
•
France A (SME)
o In vivo models of CNS disease (Parkinson’s Disease, Brain Tumor Glioblastoma, Stroke models)
o In vivo toxicity
o PK, Biodistribution studies
•
France B (ACADEMY)
o in vitro BBB (astrocyte/endothelial cell co-cultures), models of pathological BBB (inflamed BBB)
o HTS platform for the development of peptide-vectors for Np, drug and imaging agent targeting to
the CNS across the BBB via receptor mediated trancytosis (RMT).
•
France C (ACADEMY)
o Gene therapy
Delivery of siRNA targeted against the prion protein (in vitro and in vivo)
Delivery of growth factors (bdnf etc..).
in vivo and in vitro experimental models.
•
France D (SME)
o Imaging techniques
o Optical Imaging - Scintigraphy - Autoradiography
•
Portugal (SME)
o Synthesis of peptides and antibodies
o Gene therapy
•
Czech Republic (ACADEMY)
o Toxicity of Nanoparticles
o Interaction with drug metabolising enzymes
•
Denmark (ACADEMY)
o In vivo experiments on olivocerebellar degeneration and Purkinje cell death and excitotoxic model
of epileptic seizures
o Novel peptides for the engineering of the Np (15 amino acids (aa) derived from the (61 aa)
endogenous protein metallothionein (MT).
•
Israel (SME)
o Imaging
o Amyotrophic Lateral Sclerosis
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Angelo Montenero
[email protected]
0521905553
Dipartimento di Chimica G.I.A.F. – Università degli Studi di Parma
Our proposal in this study is the use of sol–gel technique to synthesize
mesoporous and nanostructured reservoirs which could be implanted
giving a sustained release of the drug molecules. The devices will be
constituted by a matrix that contains the drug dispersed or suspended in
a fine state. The mechanism, as well as the kinetics and rate of drug
release will depend upon the physicochemical properties of the drug, the
materials, and the device construction. They will be basically made of
oxides derived from Titanium, Silicon (or other metals) alkoxides. The
inner structure of the inorganic matrix will be tailored (playing with the
conditions of reaction and the relative amounts of the alkoxides, for
example) to the specific peculiarity of the substance and to the length of
the administration period.
The use of alkyl substituted alkoxides together with non-substituted
alkoxides could be useful to modify the internal network structure, which
depends on the respective rates of hydrolysis and condensation, and
therefore on steric hindrance, inductive effects (of alkyl substituents) and
functionality (number of alkoxy groups) on the central atom as well as on
catalysis and solvent.
In cooperation with some European and Mexican research centers we are
working with sol-gel devices used to release anticonvulsant drugs, such as
phenytoin and valproic acid; similar reservoirs could be employed with
drugs for Alzheimer’s treatment.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Basic Research in Advanced Innovative Nanomaterials: Applications to
the Solution of Brain Disorders
(Identifier: FP7-NMP-2007-LARGE-1)
Università Autonoma Metropolitana di Città del Messico - CEE
Two years
The aim of the group is to study the treatment of three serious
neurological disorders: Epilepsy, Parkinson’s and Alzheimer’s, using an
interbrain nanodevice to deliver the drug in front of damaged membrane
neurons, in order to obtain a potential effect and avoid the
decomposition of the molecule.
Nanostructured materials biocompatible with brain tissue will be
developed. The solids will include ordered silica (SiO2) and ordered titania
(TiO2), nanostructured manganese oxides (MnOx) cryptomelane type. Also
the sol-gel nanomaterials: SiO2, TiO2, ZrO2, HfO2, hybrid nanostructured
materials and monoliths will be prepared using a variety of alkoxides and
by thoughtfully varying synthesis conditions. All the inert matrices will be
designed using soft chemistry and will require functionalization in order
to be biocompatible with the surrounding neural membranes. The drugs
to be encapsulated within the devices include for Epilepsy, valproic acid,
sodium phenytoine, leviracetam and topiramate; for Parkinson, L-Dopa,
and dopamine. For Alzheimer the aim is to quantify the change in
concentration of Filaments tangles Helicoidally (PHFs) and
glyceraldehyde-3-phosphate dehydrogenase (GAPDH), in presence of
selective serotonine adsorbent nanomaterial.
Nome
Contatti
Istituto/Dipartimento
MASSERINI MASSIMO
0264488203
[email protected]
Dip. Medicina Sperimentale
Via Cadore 48
20052 Monza
Università Milano Bicocca
Proposta di ricerca
New nanodevices overcoming the blood-brain barrier, for the therapy of Alzheimer Disease
Nanodevices offer an attractive mean of performing therapy and diagnosis with their high potential for
multi-task functionalization. Theoretically, multi-task functionalization may confer on nanodevices a wide
array of properties, such as stealth characteristics once they enter the bloodstream (ability to avoid the
reticulo-endothelial system, RES), ability to cross the blood-brain barrier (BBB) and, of course, ability to
reach a target. In addition, since it is possible to confer on them features of biocompatibility, lack of
toxicity and immunogenicity, biodegradability, simple preparation and high physical stability, the
nanotechnology -based systems are intelligent tools for diagnostics, prognostics, and controlled and
sustained delivery of therapeutic agents to specific targets .
However, the BBB represents a formidable challenge to the delivery of therapeutic agents to the central
nervous system, and in particular for the treatment of Alzheimer disease, due to its complex, high
selectivity, to the passage of desired molecules into the brain parenchyma. This fact represents an
handicap also for testing new and future drugs. These issues has not yet been solved with the existing
nanotechnologies, and need strong investments in the future years. For this, completely new approaches
are necessary.
The proposal is to realize new nano-devices, for overcoming the BBB, for the therapy of the Alzheimer
disease. For this purpose, new approaches should be sought,, such as the possibility of nanodevices to
carry molecular factories able to perform small tasks; example are the ability to enzymatically modify the
architecture of the neuronal membrane modifying the activity of resident molecules involved in the
mechanisms of the disease.
Area di interesse identificata
Finanziamenti ricevuti
2.930.387,00€
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Nanomedicine
Nanoparticles for therapy and diagnosis of Alazheimer Disease
EC
5 y (2008-2013)
A hallmark of the disease in the AD brain is extracellular aggregates
(plaques) of the cytotoxic peptide β-amyloid (Aβ). This project intends to
use nanoparticles (NPs) for therapy and diagnosis, singly or combined
(theranostics), focusing on brain Aβ as the target. Brain and blood Aβ are
in equilibrium across the blood-brain barrier (BBB), so the project also
considers blood Aβ as a target.
Different NPs will be multiple-functionalized with: i) molecules interacting
with Aβ, ii) molecules stimulating BBB crossing, ii) PET or MRI contrast
agents. Artificial and cellular models will be used ; the efficacy of NPs will
eventually be evaluated in rodent models of AD. Different routes of
administration (i.v., oral, nasal) or different protocols (two-step, NP
cocktails, aerosols) will be employed to boost NP brain delivery. The
prediction is that NPs will range, detect, disaggregate, and remove brain
Aβ. Besides this action, NPs will interact with blood Aβ, drawing out the
excess of brain peptide by a “sink” effect.
2. PHARMACOLOGY
D-Novel Drugs
Nome
Contatti
Lamberto Maffei
[email protected]
Istituto/Dipartimento
Istituto di Neuroscienze del CNR, Pisa
Proposta di ricerca
In human subjects, building on the results obtained in subjects with Mild Cognitive Impairment (MCI) or in
the very initial stages of Alzheimer’s Disease (AD) with the ongoing project “Train the Brain”, we plan to:
extend the intervention to a second group of subjects with MCI
assess the efficacy of the training employed in “Train the Brain” in subjects in more advanced stages of AD
determine the extent of the training beneficial effects by means of extensive follow up of the subjects
included in “Train the Brain”
determine the most effective components of the training
In murine AD models, we plan to:
characterize the cellular and molecular mechanisms underlying the action of Intranasal administration (IA)
of Neurotrophins and of Environmental Enrichment (EE) in preventing the onset or in ameliorating the
progression of the disease.
develop new non invasive strategies aimed at preventing or rescuing cognitive deficits and at preventing
the onset or reducing the progression of the neurodegeneration
We shall also evaluate whether exposure to an “impoverished” environment accelerates the cognitive
decline in animal models of AD.
We have already exploited IA of NGF (De Rosa et al., 2005) to rescue visual memory deficits in a mouse
model of NGF deprivation which exhibits all hallmarks of human AD and we have recently shown in the
same model that EE prevents the onset of memory deficits and ameliorates AD hallmarks (Berardi et al.,
2007). We have also recently shown that EE and fluoxetine treatment enhances adult visual cortical
plasticity to the point of promoting recovery from defective visual development (Sale et al., 2007; Maya
Vetencourt et al., 2008; Sale et al., 2009); cortical plasticity enhancement seems highly effective in
rescuing cognitive deficits in models of neurodegeneration even in advanced stages of neuronal loss
(Fischer et al, 2007).
Area di interesse identificata
- New treatment strategies
- Translational research
Finanziamenti ricevuti
Titolo progetto
Train the brain: clinical and experimental
study of the efficacy of cognitive training and
physical exercise in dementia
Ente finanziatore
Fondazione Cassa di Risparmio di Pisa
Durata progetto
May 2009-May 2012
Abstract del progetto
Age-dependent cognitive decline is destined to become a highly
impacting problem at the clinical, economical and assistance level. Age,
indeed, is the major risk factor for dementia. In Italy, there are around
700.000 patients diagnosed with dementia and around 100.000 new
cases every year.
Among the pathologies which may lead to dementia, Alzheimer's Disease
(AD) and vascular dementia (VaD) are by far the most frequent.
At present, there are no effective therapeutic strategies for AD or VaD,
which are still pathologies orphan of treatment. It is therefore more and
more apparent the need of testing, validating and implementing new
strategies aimed at preventing and/or slowing down cognitive decline,
starting from the early stages of the disease. We propose to assess the
efficacy of a combination of physical exercise and cognitive training in
slowing down or arresting the progression of symptoms in subjects at risk
of or at the very initial stages of AD and VaD. The scientific rationale for
this proposal is described below.
a): several studies in humans have demonstrated that the exposure to a
cognitively and socially stimulating environment and physical exercise
have beneficial effects on brain functioning, especially in the elderly, and
reduce the risk of developing dementia (e.g. Laurin et. al, 2001; Fratiglioni
et al., 2004; Podewils et al., 2005; Marx, 2005; Kramer and Erickson,
2007). The estimated reduction in the risk of developing dementia varies
from study to study but generally ranges between 20 and 50%.
b) In parallel, a large number of studies performed in animal models has
demonstrated that physical exercise and expossure to a cognitive and
socially stimulating environment (a combination known as “enriched
environment”, EE) ameliorates cognitive performance, slow down the
decline associated with aging, are neuroprotective, and enhance neural
plasticity (Cotman and Berchtoldt, 2002; Nithianatharajah and Hannan,
2006). EE ameliorates cognitive deficits in animal models of AD (Adlard et
al., 2005; Jankowsky et al., 2005) and is also capable to revert cognitive
deficits when they are already well evident. These results show the EE
potential as non pharmacological therapeutic strategy not only to prevent
the onset of cognitive deficits but also to rescue them.
Studies on the efficacy of a combined physical and cognitive intervention
in subjects with dementia are very few and present methodological limits.
AIMS
Primary: To assess the efficacy of protocols of physical exercise and
cognitive stimulation on the progression of the disease in subjects at risk
of, or with AD and VaD in the initial stages of the disease, identified
through an advanced battery of diagnostic tests.
Secondary: To develop a nonpharmacological preventive/therapeutic
strategy easily applicable to humans and exploitable by the National and
Regional Health Service and to study the underlying cellular and
molecular mechanism of action in animal models.
Nome
Prof. Aldo Andreani
Prof. Aldo Roda
Contatti
Istituto/Dipartimento
[email protected]
2099734
Tel. +39 051 2099714 Fax: +39 051
[email protected]
343398
Tel. +39 051 343398
Fax: +39 051
Dipartimento di Scienze Farmaceutiche - Università di Bologna
Via Belmeloro 6 40126 Bologna
Proposta di ricerca
Synthesis and screening of new AChE/BuChE inhibitors
The cholinergic transmission in the central nervous system is important for the regulation of memory
and learning process. Numerous studies have highlighted that the cholinergic neurotransmission
system is profoundly compromised in the brain affected by Alzheimer’s disease (AD), with losses of
cholinergic neurons and synapses occurring in forebrain, cortex, and hippocampus. The efficacy of
cholinergic therapies in AD supports the cholinergic hypothesis and validates this neurotransmitter
system as a therapeutic target. Donepezil inaugurated a new class of acetylcholinesterase (AChE)
inhibitors with longer and more selective action with manageable adverse effects. It binds both the
active-site gorge and the peripheral site through benzylpiperidine and indanone moieties.
In an attempt to obtain analogues of donepezil that could maintain its main spatial and
physicochemical characteristics while being more compact and less flexible, we applied a strategy to
synthesize and rapidly evaluate a series of molecules to be eventually developed as AChE inhibitors
for use in AD. We designed a small library of benzylpiperidinone derivatives, where the indanone
group was replaced by a bioisosteric indole or pyrrole ring bearing different substituents. The
biochemical evaluation of the newly synthesized series was performed in collaboration with the
research group coordinated by Prof. Aldo Roda by using a chemiluminescent (CL) method suitable for
high-throughput screening (HTS) of AChE inhibitors [1]. The new derivatives showed a rather low
degree of potency against the enzyme compared to donepezil, but the molecular skeleton allows for
structural modifications which could be done to explore the potential of the series toward new
cholinergic agents useful in the treatment of AD [2].
To give a further insight into cholinesterase inhibitors, we also prepared new imidazo[2,1-b]thiazole
derivatives [3,4], which were also subjected to CL screening for the evaluation of AChE and
butyrylcholinesterase (BuChE) inhibitory activity. These studies allowed the identification of some
analogues displaying a peculiar AChE-selective inhibitory activity in the micromolar range [4].
Both AChE and BuChE hydrolyze ACh, albeit with slightly different kinetics, and coexist ubiquitously in
humans. BuChE is widely distributed in the body of vertebrates and is implicated in neurogenesis and
regulation of cell proliferation and differentiation. AChE and BuChE inhibitors could reverse the
cognitive deficits associated with damaging of cholinergic neurons. These enzymes are encoded by
different genes and clearly differ in substrate specificity and sensitivity to inhibitors, likely due mainly
to structural differences in the active site gorge. In brain affected by severe AD, the reduction of
AChE is accompanied by an up to 2-fold increase in BuChE levels. Moreover both enzymes are
involved in aggregation of beta-amyloid and in formation/maturation of senile plaques, therefore the
use of reversible cholinesterase inhibitors is considered an attractive therapeutic approach. Unlike
AChE, the physiologic function of BuChE in normal and diseased humans is not completely clear yet,
although a role in neurodegenerative disorders has been suggested. Ongoing elucidation of the
properties and behavior of BuChE in normal and AD brains supports a role for BuChE in AD pathology
as well as normal cognition. In AD clinical studies the cognitive improvement was correlated with
rivastigmine (Exelon) treatment. This drug is a well known AChE/BuChE inhibitor and these studies
support a role for central BuChE in addition to AChE inhibition in modulating cholinergic function.
Observation made in these studies indicates that a good balance of inhibition of both AChE and
BuChE may be beneficial in treating the cognitive decline in AD.
The research we propose concerns the design and synthesis of small libraries of molecules, whose
AChE/BuChE inhibitory activity will be assayed using the HTS CL method for AChE/BuChE inhibitors
described above. The availability of a simple, rapid, and sensitive HTS CL method will facilitate the
study of the biological activity of the designed compounds and the identification of the most
promising molecules. In addition, the effect of AChE inhibitors could be evaluated “ex vivo” in
experiment animals by ultrasensitive CL microscope imaging. In fact, the CL system can be also
employed for the localization and quantification of AChE in brain tissue sections [5].
Overall, these studies could furnish new molecular tools useful in elucidating structural and
functional differences between AChE and BuChE and may contribute to the development of new
potential drugs for the treatment of AD.
[1] Guardigli M., Pasini P., Mirasoli A., Leoni A., Andreani A., Roda A.: Chemiluminescent highthroughput microassay for evaluation of acetylcholinesterase inhibitors. Anal. Chim. Acta, 535, 139144, 2005.
[2] Andreani A., Cavalli A., Granaiola M., Guardigli M., Leoni A., Locatelli A., Morigi R., Rambaldi M.,
Recanatini M., Roda A.: Synthesis and screening for antiacetylcholinesterase activity of (1-benzil-4oxopiperidin-3-ylidene)methylindoles and –pyrroles related to donezepil. J. Med. Chem. 44, 40114014, 2001.
[3] Andreani A., Granaiola M., Guardigli M., Leoni A., Locatelli A., Morigi R., Rambaldi M., Roda A.:
Synthesis and chemiluminescent high throughput screening for inhibition of acetylcholinesterase
activity by imidazo[2,1-b]thiazole derivatives. Eur. J. Med. Chem., 40, 1331-1334, 2005.
[4] Andreani A., Burnelli S., Granaiola M., Guardigli M., Leoni A., Locatelli A., Morigi R., Rambaldi M.,
Rizzoli M., Varoli L., Roda A.: Chemiluminescent high-throughput microassay applied to imidazo[2,1b]thiazole derivatives as potential acetylcholinesterase and butyrylcholinesterase inhibitors. Eur. J.
Med. Chem., 43, 657–661, 2008.
[5] Pasini P., Musiani M., Russo C., Valenti P., Aicardi G., Crabtree J.E., Baraldini M., Roda A.:
Chemiluminescence imaging in bioanalysis, J. Pharm. Biomed. Anal., 18, 555-564, 1998.
Area di interesse identificata
Multidisciplinary projects
Nome
Contatti
Istituto/Dipartimento
Olga Bruno
[email protected]
Tel: +39 010 353 8367
Dipartimento di Scienze Farmaceutiche – Università degli Studi di
Genova
Viale Benedetto XV,3 16132 Genova
Proposta di ricerca
In the past few years, it has been suggested that dysfunctions of the cAMP/PKA/CREB pathway could be a
major cause of the cognitive deficits characterizing Alzheimer Disease (AD). Indeed, in vitro experiments
have shown that Aβ42 is able to reduce the function of this pathway in hippocampal neurons, thus leading
to impairment of long term potentiation (LTP) and both effects can be reversed by rolipram, a well known
PDE4 inhibitor. More interestingly, rolipram has been shown to restore LTP and to ameliorate cognitive
deficits in a double transgenic (APP/PS1) murine model of AD. Recently, sub-chronic rolipram treatment
was reported to improve long-term memory performances also in normal rodents.
Despite PDE4 inhibitors have been indicated as promising therapeutical agents for AD, rolipram cannot be
clinically used due to its adverse effects, particularly sedation and nausea. Among the different PDE4s,
PDE4D is highly expressed in hippocampal regions and some isoforms seem to be augmented in the
hippocampus of AD patients.
Recently we have synthesized a series of innovative selective PDE4D2, PDE4D3 inhibitors,1 which increase
cAMP levels in the hippocampus and improve cognitive functions in preliminary in vivo pre-clinical studies.
Surprisingly, in vitro experiments showed that our compounds, as well as rolipram, augment Aβ levels in
neuronal cultured cells, a result in agreement with a recent study showing that dimebolin, a drug known to
ameliorate cognitive deficits in aged rodents and in people suffering from mild AD, causes an acute
increase of Aβ brain levels in experimental animals.
Our project is aimed at studying new PDE4D selective inhibitors as novel therapeutic agents in AD. These
compounds could be also useful pharmacological tools to increase the knowledge on the role of different
PDE4 isoforms in CNS under physiologic as well as pathologic conditions, and to verify the relationship
between PDE4D inhibition, A levels and memory improvement.
Our research unit skilful in medicinal chemistry will work to modify the chemical structure of previously
synthesized compounds in order to increase its selectivity, potency and in vivo activity. Our first goal is to
obtain a beneficial effect on memory performances without emesis.
The molecular modifications will be performed basing on a preliminary computational study aimed to
show what structural features could discriminate between different PDE4D splicing variants. Thus docking
studies and 3D-QSAR analysis will be used to filter the large number of new molecular entities deriving
from various chemical modifications on our previously synthesized compounds.
1) Bruno O, Romussi A, Spallarossa A, Brullo C, Schenone S, Bondavalli F, Vanthuyne N, Roussel C. New
selective phosphodiesterase 4D inhibitors differently acting on long, short, and supershort isoforms. J Med
Chem. 2009 52(21), 6546-57.
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Drug Design and synthesis of therapeutic agents for neurodegenerative
disorders, particularly for Alzheimes Disease.
Heterocycle compounds as neuroprotective agents
(MIUR, Italian Minister of University Research) 2005032713_005
Ronsisvalle Giuseppe (Principal Investigator)
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Bruno Olga Role: Co-investigator
1/01/06-31/12/07
This study was aimed at synthesizing new molecules acting by different
pathways in neurodegenerative pathologies.
Heterocycle compounds as neuroprotective agents
(MIUR, Italian Minister of University Research) 2007E8CRF3_003
Ronsisvalle Giuseppe (Principal Investigator)
Bruno Olga Role: Genoa Unit PI
22/09/08-22/09/10
This grant is a renewal of previously financial support for a large study
aimed to synthesize new molecules acting by different pathways in
neurodegenerative pathologies.
Nome
Contatti
Istituto/Dipartimento
Fabrizio Tagliavini
Tel +39 02 2394 2260; Fax +39 02 2394 2101
Email: [email protected]
Sito web: http://www.istituto-besta.it
UO Neuropatologia – Neurologia 5
Fondazione IRCCS Istituto Neurologico “Carlo Besta”
Via Celoria ,11 - 20133 Milano- ITALY
Proposta di ricerca
The research group: The Division comprises (i) a Clinical Unit (Dementia Center) devoted to the
diagnosis and treatment of patients with degenerative dementias (over 2000 out-patients and 150 inpatients per year) and (ii) a Laboratory Unit (certified ISO 9001:2000 - Registration Number 26080,
dedicated to the analysis of genes and biomarkers associated with degenerative dementias, postmortem characterization of the disease process and disease-specific protein, and experimental
studies on disease pathogenesis and the development of therapeutic strategies. The course of action
of this activity can be expressed as a “Bed to Bench to Bed” cycle, with the ultimate goal to identify
disease-modifying drugs in preclinical settings and apply them to patients. In this regard, the Clinical
Unit is currently coordinating a multicentre phase II clinical trial supported by AIFA, to test the
effectiveness of an anti-amyloidogenic molecule identified in experimental models. This
comprehensive approach has also the great advantage to collect biological samples (plasma, DNA, CSF
and brain tissue) from fully characterized patients. In this regard the Laboratory Unit is part of the
Network of Excellence “BrainNet” Europe devoted to biobanking, harmonization of assessment tools
and standardization of diagnostic criteria of neurodegenerative diseases.
The permanent staff of the Division is composed by 8 MD and two technicians. In addition, the staff
comprises 13 post-doctoral fellows (7 PhD, 3 MDV, 1 MD, 1 psychologist), 2 PhD students and 1
technician who are committed to the research activities on degenerative dementias with different
expertise and roles. On the overall, the team has large experience in a variety of techniques spanning
from neuropathology (including immunohistochemistry, morphometry, electron microscopy and
atomic force microscopy), to molecular genetics, biochemistry (purification and characterization of
disease-specific proteins), cellular and molecular biology, and animal models. The Division of
Neuropathology is provided with fully equipped laboratories for histology, immunohistochemistry,
electron microscopy, atomic force microscopy, biochemistry, and molecular and cellular biology, and
a 60 sqm BL3 facility. Moreover an animal facility composed by a 140 sqm environmental controlled
area, provided with a HVAC system and heap-filtered ventilated racks, with separate rooms for
maintaining and breeding mouse lines, and carrying out surgical procedures, collection of samples
and post-mortem examination is available.
B. Preclinical research
• Recessive A673V APP mutation: molecular mechanisms and development of a new therapeutic
strategy for sporadic AD
We have recently identified an APP mutation (A673V) that causes early-onset AD only in the
homozygous state while the heterozygous carriers are not affected. This mutation strongly boosts the
production and amyloidogenic properties of Aβ. However, the interaction of A673V-mutated and
wild-type peptides inhibits amyloidogenesis and Aβ-mediated neurotoxicity (Di Fede et al., Science
2009, 323:1473-7). These findings are consistent with the observation that the A673V heterozygous
carriers do not develop disease and offer grounds for a novel therapeutic strategy based on modified
Aβ peptides. A research priority of our lab is to unravel the molecular mechanisms of the opposite
effects of the A673V APP mutation in homo- or heterozygous state on amyloidogenesis and to develop
a lead compound for AD therapy based on A673V-modified Aβ peptides or peptido-mimetic
molecules. To accomplish these objectives we have generated a panel of transfected cells and
transgenic C. elegans expressing human APP or Aβ with the A673V mutation, respectively, and
transgenic mouse lines expressing A673V-mutated APP in the homozygous or heterozygous state.
Furthermore, we have identified a prototypic lead compound corresponding to a six-mer Aβ peptide
with the A673V substitution and are currently working on brain delivery systems.
Area di interesse
identificata
CLINICA/PRECLINICA
-
Genetic susceptibility to Alzheimer’s disease (AD)
Early diagnosis
Biomarkers
Developing competitive animal models to study AD
Studying early onset forms of AD to follow-up disease progression
Cinical trials with or without involvement of pharmaceutical companies
Basic research
Biobanking (blood sample, CSF, brain repository…)
Standardization of diagnostic criteria
New treatment strategies
- Translational research
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Simone OTTONELLO
+39 0521 905646 (office)
+39 0521 905148 (lab)
[email protected]
Department of Biochemistry and Molecular Biology
Viale G.P. Usberti 23/A
University of Parma
43100 Parma (Italy)
-“Biomarkers” / “Early diagnosis”
Implementation of high-resolution techniques for Aoligomer detection
and quantification in brain samples from Tg-mouse models of Alzheimer
disease and in body fluids, including human CSF.
-“Basic research”
Yeast mutant strains (selected from a genome-wide mutant collection)
conditionally expressing Alzheimer-related pathogenic polypeptides as
tools to unravel new fundamental aspects of AD pathogenesis (e.g.,
mitochondrial function impairment) and potential treatments thereof.
-“New treatment strategies”
Second generation, recombinant A peptide immunogens as prototype
vaccines for AD treatment/prevention (active immunization approach)
and as primary antigens for the production of conformation-sensitive
therapeutic monoclonal antibodies (passive immunization approach).
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
“Novel anti-A immunization approaches for the treatment of Alzheimer
disease”
Pharmaceutical Company
Durata progetto
2 years
Titolo progetto
Ente finanziatore
“Further studies on the Trx-A antigen and its potential as an anti-AD
vaccine”
Pharmaceutical Company
Durata progetto
year
Titolo progetto
Ente finanziatore
“Target selectivity of NSAID -secretase modulators evaluated by oligomacroarray analysis”
Pharmaceutical Company
Durata progetto
year
Titolo progetto
"Large-scale production of recombinant TrxA for a preclinical study in
Ente finanziatore
Durata progetto
Titolo progetto
Ente finanziatore
Durata progetto
Tg-AD mice”
Pharmaceutical Company
months
"Immunoelectrophoretic analysis of A oligomers in brain samples from
Tg-AD mice treated with a NSAID -secretase modulator”
Pharmaceutical Company
6 months (ongoing)
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
MAURIZIO SCARPA
Area di interesse
identificata
Finanziamenti ricevuti
Titolo progetto
PEDIATRIC NEURODEGENERATIVE DISEASES
[email protected]
DEPARTMENT OF PEDIATRICS UNIVERSITY OF PADOVA ITALY
Ente finanziatore
BRAINS4BRAIN – Treating Pediatric Neurodegenerative Diseases:
From Laboratory Bench to Bedside
EUROPEAN SCIENCE FOUNDATION
Durata progetto
Exploratory Workshop March 3-5 2010 Frankfurt
Abstract del progetto
Neurodegenerative diseases are most prevalent in the elderly, but can in rare cases also
affect individuals early in life. In children, neurodegeneration leads to severe mental
retardation and premature death with devastating consequences on their immediate
environment and relatively high costs for society. Within the EU pediatric
neurodegenerative disorders are considered “Rare”. Because of the low-prevalence of
these disorders, there is often a striking lack of information, research, treatment and
expert availability. There is also often a significant delay before a definitive diagnosis is
achieved. BRAINS4BRAIN calls for a collaborative research effort to focus the available
multidisciplinary European brainpower with relevance to tackling pediatric
neurodegenerative diseases. BRAINS4BRAIN (www.brains4brain.eu) is a European task
force aimed at developing and implementing new therapies for pediatric
neurodegenerative diseases for which, at the moment, no effective therapy is available.
The task force wishes to create a coordinated European effort towards i) the
comprehension of pathophysiology processes of these pediatric neurological disorders, ii)
the implementation of knowledge on the BBB, iii) the development of new molecular
and/or biochemical strategies to overcome the BBB for brain-targeted drug delivery, and
iv) the development and implementation of diagnostics and clinical therapies to timely
detect and treat these devastating CNS disorders. Importantly, it is foreseen that such
knowledge and insights will also be valuable for understanding and treating other
important neurological diseases such as Alzheimer’s, Parkinson’s Disease, and epilepsy.
LSDs are metabolic disorders, caused by the lack of certain (lysosomal) enzymes or
lysosome components, thus preventing the complete degradation of macromolecules and
the recycling of their components.
The accumulation of intermediate degradation products affects the appropriate
functioning of lysosomes and other cellular organelles.
Accumulation starts immediately after birth and progressively worsens, often affecting
several organs, including the Central Nervous System (CNS). CNS pathology causes mental
retardation and progressive neurodegeneration that ultimately ends in early death of
these young patients. LSDs are the only group of pediatric neurodegenerative diseases
for which therapy that can reverse the natural history of the disease in peripheral organs
is available (Enzyme Replacement Therapy, ERT).
Unfortunately, ERT is currently unable to effectively reach the CNS to stop the lethal
progression of the neurodegeneration. Nonetheless, ERT in combination with i) the
advanced knowledge of the (genetic- and biochemical) causes for the development of
neurodegeneration in LSDs and ii) the availability within Europe of well established in
vitro and animal models, now provides the unique opportunity to decipher the cascade of
events leading to loss of brain plasticity and mental retardation, as well as its possible
reversal. For instance, suppressing the primary cause of neurodegeneration (e.g. by ERT)
in a young brain –that at this stage of development retains considerable plasticity–
maximizes the potential for neurological repair. Important to note is that common
secondary events in both pediatric and adult neurodegenerative diseases lead to
neurodegeneration. Studies on LSDs therefore have the ability to provide unique insights
into the pathophysiology and restorative capacities of neurodegenerative diseases in
general. Therapies based on ERT can modify the natural history of some LSDs in the
peripheral organs (liver, spleen, joint mobility etc.). However, the BBB prevents the
therapeutic enzymes used from reaching the CNS and modifying the course of
neurodegeneration 1. This limitation can be circumvented by non-pharmaceutical therapy
in which sources of enzyme (e.g. a viral vector) are created by surgical injection directly
into the CNS. It might eventually also be achieved to some extend by pharmaceutical
therapies based on BBB traversing small molecules aimed at reducing the accumulation of
molecules requiring degradation (e.g. Substrate Reduction Therapy (SRT):inhibiting early
enzymes in the biosynthetic pathway, or Chemical Chaperones: assisting the affected
enzyme to attain its correct, active conformation2).3,4 However, ERT is likely to be the
most efficient strategy with the least side effects. As neurodegeneration in LSDs affects
the entire brain, the technological challenge is to safely deliver ERT efficiently to all
affected areas. This might be achieved only by developing strategies enabling the
therapeutic enzyme to cross the BBB. Tools to enable the entry of curative molecules
across the BBB into the CNS will represent a major breakthrough for the long-term
therapy of many (if not all) CNS diseases. Several cutting edge initiatives using different
approaches to specifically deliver molecules across the BBB (e.g. exploiting certain
transporters or manipulating BBB permeability) exist within Europe.
Nome
Contatti
Paola Tirassa
[email protected]; INMM- CNR Istituto di Neurobiologia e Medicina
Molecolare Via del Fosso di Fiorano, 64 00143 ROMA ITALY Phone: + 39
06 501 703 236 Fax: + 39 06 501 703 313
Istituto/Dipartimento
Proposta di ricerca
Institute of Neurobiology and Molecular Medicine CNR
2) Basic research : New treatment strategies based on Neurotrophins
f) Ocular NGF administration as a novel non invasive approach to protect brain-NGF target neurons that
degenerate in Alzheimer’s disease (L. Aloe, P. Tirassa). Nerve growth factor and (NGF) is a soluble protein
that plays a protective role on brain neurons that degenerate in age-related brain disorders, including
Alzheimer’s disease (AD). We have shown that conjunctively applied NGF can protect injured brain
neurons and damaged retinal ganglion neurons suggesting that topical eye NGF application might be a
novel alternative for delivering NGF into the brain and for protecting brain neurons and reducing memory
deficits occurring during early events of AD and glaucoma. This hypothesis is currently actively under
investigation in collaboration with other research groups.
Lambiase A, Pagani L, Di Fausto V, Sposato V, Coassin M, Bonini S, Aloe L. Nerve growth factor eye drop
administrated on the ocular surface of rodents affects the nucleus basalis and septum: biochemical and
structural evidence. Brain Res. 2007;1127:45-51.
Di Fausto V, Fiore M, Tirassa P, Lambiase A, Aloe L. Eye drop NGF administration promotes the recovery of
chemically injured cholinergic neurons of adult mouse forebrain. Eye drop NGF administration promotes
the recovery of chemically injured cholinergic neurons of adult mouse forebrain. Eur J Neurosci.
2007;26:2473-80.
Lambiase A, Aloe L, Centofanti M, Parisi V, Mantelli F, Colafrancesco V,
Manni GL, Bucci MG, Bonini S, Levi-Montalcini R. Experimental and clinical evidence of neuroprotection by
nerve growth factor eye drops: Implications for glaucoma. Proc Natl Acad Sci U S A, 2009; 109: 1346913474,.
g) Electro-acupuncture and polyphenols as novel approaches to reduce neurodegeneration ( L Manni, M
Fiore). Preliminary data indicate that diabetes induced in adult rats by injection with streptozotocin causes
a decrease of brain NGF and a concomitant increase of tau phosphorylation, that were all counteracted by
low-frequency electro-acupuncture (EA). Using molecular and behavioural approaches the research
project will be aimed at: i) Investigate the possible link between diabetes, brain NGF presence/activity and
the development of AD-associated pathological features; ii) Study the effect of EA on the above mentioned
diabetes-associated brain dysfunction. Alcoholism is related to neurodegeneration and diabetes. The use
of polyphenols extracted by olive oil, olive leaves and olive seeds or by grapes (Fiore et al 2009) to prevent
or limit mouse brain neurodegeneration due to aging or ethanol consumption will be evaluated.
Manni L, Aloe L, Fiore M. Changes in cognition induced by social isolation in the mouse are restored by
electro-acupuncture. Physiol Behav. 2009;98(5):537-42
Fiore M, Laviola G, Aloe L, di Fausto V, Mancinelli R, Ceccanti M. Early exposure to ethanol but not red wine
at the same alcohol concentration induces behavioral and brain neurotrophin alterations in young and
adult mice. Neurotoxicology. 2009;30(1):59-71.
h) Human neural stem cells lentivirally transduced with human BDNF (C. Cenciarelli, P. Casalbore). A
strong body of evidences show that BDNF result critically involved in Alzheimer’s and Huntington’s
diseases. Recently, it has been shown that BDNF knockdown within NSC abolishes the cognitive benefits of
NSC delivery. The aim is to use propagating human neural stem cells (hNSC) lentivirally transduced with
human BDNF as cellular therapy for replacing degenerating neurons in disease, trauma and toxic insults.
Cenciarelli C, Budoni M, Mercanti D, Fernandez E, Pallini R, Aloe L, Cimino V, Maira G, Casalbore P. In vitro
analysis of mouse neural stem cells genetically modified to stably express human NGF by a novel multigenic
viral expression system. Neurol Res. 2006, 28:505-12.
Casalbore P, Barone I, Felsani A, D’Agnano I, Michetti F, Maira G and Cenciarelli C. Neural stem cells
modified to express BDNF antagonize trimethyltin-induced neurotoxicity through PI3K/Akt and MAP kinase
pathways . Accepted on J. Cell. Physiol. 2010.
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Micaela Morelli
Department of Toxicology, University of Cagliari
New symptomatic and neuroprotective therapies for Parkinson’s
disease
1) Development and dissemination of innovative methodologies
for planning, synthesis and determination of biological activity
of new ligands for membrane receptors (FIRB:RBNE03YA3L_004)
2) Evaluation of behavioural and biochemical changes in
'Engrailed' mutant mice (PRIN 2006)
3) : Evaluation of neuroprotective effects of ST1535 in animal
model of Parkinson ‘s disease (SIGMA-TAU industry)
1-2) MURST
3 ) SIGMA-TAU Industry
1) 3 years
2) 2 years
3) 1 year
1) To determine the in vivo and ex vivo biological activity of
adenosine A2A antagonists compounds of new synthesis. Three
compounds symptomatically active on models of Parkinson’s
disease have been selected on the basis of the results obtained.
Moreover their potential neuroprotective effects has been
recently investigated in models of Parkinson’s disease.
2) The engrailed strain of mice has shown behavioural deficits and
biochemical changes superimposible of Parkinson’s disease
models. This strain of mice is now used as a model of this disease.
3) Evaluation of neuroprotective and anti-inflammatory effects of
new adenosine A2A receptor antagonist ST1535 in animal model
of Parkinson ‘s disease. ST1535 has shown positive symptomatic
improvement in motor functions and also neuroprotective and
anti-inflammatory effects in the MPTP mouse model of
Parkinson’s disease.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Proposta di ricerca
Maria P. Abbracchio
Dept Pharmacol Sci, Univ Milan, Italy
+39-02-50318310/355
[email protected]
Area di interesse identificata
Innovative neuro-reparative strategies via the implementation of
endogenous neurogenesis and gliogenesis: focus on the new P2Y-like
GPR17 receptor
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
PURINOCEPTORS AND NEUROPROTECTION: FOCUS ON THE NEW
PURINERGIC RECEPTOR GPR17
COFIN-MIUR 2008;
Durata progetto
2 anni
Titolo progetto
Ente finanziatore
IMPLEMENTATION AND GLIOGENESIS VIA THE PURINERGIC SYSTEM: A
NEW STRATEGY TO REPAIR ACUTE NEURODEGENERATIVE DISEASE
Ministero della Salute 2007
Durata progetto
2 anni
Abstract delle ricerche in Deciphering the molecular mechanisms controlling the behaviour of
corso
endogenous neural stem cells is of paramount importance to understand
the processes at the basis of brain recovery and to foster the brain’s
ability to repair itself in acute and chronic neurodegenerative diseases.
We have recently identified a new P2Y-like G-protein-coupled
purinoceptor GPR17, activated by both uracil nucleotides and cysteinylleukotrienes (cysLTs) (Ciana P et al., The orphan receptor GPR17 identified
as a new dual uracil nucleotides/cysteinyl-leukotrienes receptor. EMBO J
25:4615-27, 2006). We have established a national consortium (the
GPR17 research team) that specifically studies the role of this receptor in
cerebral diseases. Initial data in rodent models of focal brain ischemia
have shown that GPR17 participates to both the propagation of brain
damage and to the subsequent post-injury remodelling and repair (Lecca
D et al., The recently identified P2Y-like receptor GPR17 is a sensor of
brain damage and a new target for brain repair. PLoS ONE 3:e3579, 1-15,
2008). Specifically, GPR17 was found to activate the quiescent
oligodendrocyte precursor cells that are still present in the adult brain,
inducing them to resume myelination, thus re-establing neuron-toneuron communication. GPR17 was also found to be expressed by the
ependymal cells lining spinal cord’s central canal (Ceruti S et al., The P2Ylike receptor GPR17 as a sensor of damage and a new potential target in
spinal cord injury. Brain 132:2206-18, 2009) that are believed to
represent the true adult stem cells in this part of the central nervous
system. Upon spinal cord mechanical injury, these cells started
proliferating and re-expressing markers of multipotent stem-like cells
(ibidem). Interestingly, in the adult brain, GPR17 specifically decorates a
precursor cell (NG2-positive neural precursors) that normally
differentiates to oligodendrocytes, but that, under appropriate
conditions, can be addressed to generate functional new neurons and
new astrocytes. These data make GPR17 an interesting new target not
only for stroke but also for chronic human neurodegenerative diseases
characterized by demyelination or neuronal pathology, such as multiple
sclerosis (MS) and Alzheimer’s disease (AD). Preliminary data in animal
models indeed support this hypothesis: activated GPR17-expressing adult
precursor cells were found to accumulate, respectively, around
demyelinated wounds in an Experimental Autoimmune Encephalomyelitis
(EAE) MS animal model, and around brain plaques in a transgenic murine
model of AD (APPPS1 mice).
Our current efforts aim at:
1. We want to develop an in silico tridimensional pharmacophore model
of GPR17 to screen large virtual chemical libraries and to design new
specific ligands. The newly synthesized agonists/antagonists (including
new “dual” ligands acting at both the nucleotide and the cysLT binding
sites) will be tested in our established in vitro assays and then addressed
to their in vivo validation
2. “Old” and new GPR17 ligands will be used to unveil the exact role of
the receptor during oligodendrocyte differentiation and myelination, in
both in vitro and in vivo models, and to identify the factors regulating its
transcription in precursor cells.
3. To set the basis for a possible wider exploitment of GPR17 in adult
neurogenesis/gliogenesis, by using both isolated precursor cells and
neurospheres from control and diseased brains, we shall also develop
pharmacological protocols to address GPR17-positive precursor cells to
neurons or astrocytes.
4. The role of GPR17 in the EAE MS model will be studied in detail. Initial
information on the changes of GPR17 expression in these models will be
obtained via an in silico analysis of microarray data; then, the regional
localization, cell types, and kinetics of GPR17 expression during disease
evolution will be assessed. Finally, we shall determine the effects of a
“preventive” or “therapeutic” in vivo administration of GPR17 ligands on
animals’ weight, neurological score and disease course.
Globally, results will contribute to finding new GPR17-based regenerative
medicine approaches for human neurodegenerative diseases, allowing for
substantial improvement of “endogenous neural stem cell”-based in vivo
therapy.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Mariagrazia Grilli/Pier Luigi Canonico
Mariagrazia Grilli
0321375828 3280587577
[email protected]
DiSCAFF, University of Piemonte Orientale, Novara Italy
Proposta di ricerca
Area di interesse
identificata
Novel drugs (D).
“Pharmacological modulation of adult neurogenesis”
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Pharmacological modulation of endogenous neural stem cells for
amelioration of cognitive impairment in neuropsychiatric
disorders
Cariplo, Regione Piemonte, PRIN, Fondazione Comunità del Novarese
Durata progetto
3 anni
Abstract del progetto
As of today, the pharmacological treatment of cognitive deficits associated with
several neuropsychiatric disorders is still very limited. The essential requirement
for an ideal anti-dementia drug would be the ability to re-organize the neuronal
network in the injured brain so as to prevent/slow down the neurodegenerative
process and possibly to partially recover brain function. Structural and functional
plasticity, often referred to as “neuroplasticity”, a fundamental property of the
brain, comprises chemical, electrical, molecular and cellular responses by which
connections within a brain region and/or between brain regions are reorganized. It
is maximal during brain development but it persists to a relevant extent in
adulthood, conferring to neuronal networks a higher degree of circuit adaptation
to the novel experiences we are exposed to for our entire life. Neuroplasticity in
adulthood is in fact the neurobiological substrate of learning and memory. A
better understanding of molecular events underlying neuroplasticity may lead to
innovative therapeutic approaches in a variety of neuropsychiatric disorders
characterized by learning and memory deficits.
Different forms of neuroplasticity are known to exist in the adult brain,
including neurogenesis. A large body of experimental work has recently
established that newborn neurons continue to be added to discrete regions of the
adult nervous system including the hippocampus, an area implicated in learning
and memory which form the basis for complex behaviour patterns in vertebrates.
Several studies support the hypothesis that experience- or activity-dependent
integration of new neurons into hippocampal pre-existing neural networks
represents a functional mechanism of brain plasticity subserving specific types of
learning and memory functions. Several human disorders affecting memory and
cognition, including neurodegenerative and psychiatric disorders, are
characterized by abnormalities in synaptic structure/number as well as in
neurogenesis, including Alzheimer’s disease (AD). The present project aims to
investigate whether the pharmacological manipulation of neuroplasticity in
pathological conditions may be effective in modulating different memory stages
and systems. This would be a starting point for developing and testing novel
pharmacotherapies for cognitive disorders. In particular, the contribution of NFkappaB proteins to this process and in cognitive disorders is under investigation.
Indeed a vast body of information points at the role of the NF-kappaB family in
synaptic plasticity, learning and memory and at neuropychiatric disorders arising
when it is deranged (Grilli & Meneghini, in press, 2010). Recently, we showed
that NF-kappaB p50KO mice displayed a very selective defect in hippocampal dependent short-term memory which correlated with the disruption of
hippocampal adult neurogenesis (Denis-Donini et al., 2008). More specifically, in
the absence of p50 protein, defective neurogenesis resulting in memory defects
appeared to possibly occur at the transition between distinct maturation steps in
newly generated neurons. Intriguingly, this is the stage characterized by high
synaptic plasticity when in the adult newborn neurons with extended axons and
dendrites have to be selected so as to become integrated into the preexisting
functional network. Morover, we were also able to show that several drugs
promoting in vitro and in vivo neurogenesis actually involve the NF-kappa
signalling pathway. Altogether, these data identify NF-κB signaling components
as critical mediators of adult neurogenesis and suggest that that they may
represent previously undescribed therapeutical targets for neuropsychiatric
disorders, including AD. The overall goal of this research proposal is evaluate
whether a disregulated NF-kappaB signalling may affect mechanisms regulating
neuronal maturation and differentiation of adult generated neurons and, as a
consequence, contribute to cognitive impairment associated with relevant CNS
disorders. To this aim, we will use a relevant animal model of AD such as the
TgCRND8 mouse line as well as the p50KO mice, both characterized by cognitive
impairment. Experimental strategies will be applied to allow to prove the concept
that this signaling pathway can be pharmacologically modulated to promote adult
neurogenesis and, in turn, ameliorate cognitive impairment. Our project has the
potential to increase our understanding of the molecular participants in the
modulation of adult neurogenesis in physiology and pathology and it also holds
the potential for identifying novel strategies aimed at ameliorating cognitive
impairment associated with CNS disorders of high medical need but with no or
unsatisfactory therapy.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Fabrizio Chiti
Dip. di Scienze Biochimiche, Univ. di Firenze, V.le Morgagni 50, Firenze.
Tel: +39-055-4598319
e-mail: [email protected]
Dip. di Scienze Biochimiche, Università di Firenze, Viale Morgagni 50,
Firenze
We intend to focus on the relationship structure-function of oligomers of
the β amiloid peptide and its analogues. We will use a number of
methodologies to gain structural information on the oligomers at the
molecular level, including site-directed mutagenesis, site-directed
labelling with molecular probes to be followed in fluorescence, NMR and
EPR. We will also use cultured cells and animal models, evaluating the
effect of structural changes of the oligomers on their toxicity, of inhibitors
of toxicity such as chaperones and small molecules, of changes of the
membrane lipid content, of the presence of glycosaminoglycans and
other components of the extracellular matrix, and of protective agents
such as extrogens, IGF-1 and others that mediate neuroprotection via the
activation of the seladin-1 gene. We will use common cellular cultures but
also cultures of primary neurons of rat and human ippocampi to evaluate,
for example, interactions between oligomers and synapses or oxidative
stress of the membrane. We will also use animal models such as
transgenic mice. The following research groups will participate:
Prof. Fabrizio Chiti
Dipartimento di Scienze Biochimiche
Prof. Massimo Stefani
Dipartimento di Scienze Biochimiche
D.ssa Cristina Cecchi
Dipartimento di Scienze Biochimiche
Prof. Fiorella Casamenti
Dip. di Farmacologia Precl. e Clinica
Prof. Alessandro Peri
Dipartimento di Fisiopatologia Clinica
The role of cholesterol in the pathogenesis of Alzheimer's disease
Fondazione Cassa di Risparmio di Pistoia e Pescia and Centro
Internazionale delle Malattie neurodegenerative (CIMN)
12 months (January 2010 - December 2010)
We will investigate the relationship between membrane cholesterol
content and cytotoxicity induced by Aβ oligomers on human
neuroblastoma cells and primary fibroblasts from familial AD patients
bearing genetic mutations.
Cytotoxicity of amyloid aggregates in neuronal cells and in Alzheimer
fibroblasts: effect of putative anticytotoxic molecules.
MIUR and Università di Firenze (cofinanziamento PRIN 2008)
24 months (March 2010 – March 2012)
We will study the ability of antioxidant compounds, such as glutathione
thioesters (S-acyl-GSH) derivatives, to prevent the degenerative effects
triggered by Aβ accumulation on primary cortical neurons, human
neuroblastoma cells and primary skin fibroblasts from familial AD
patients.
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Effect of the extracellular matrix on the formation, stability and
cytotoxicity of amyloid fibrils and their oligomeric precursors
MIUR and Università di Firenze (cofinanziamento PRIN 2008)
24 months (March 2010 – March 2012)
We will study the effect of components of the extracellular matrix where
amyloid accumulate (chaperones, glycosaminoglycans, collagen fibres,
etc.) on the formation, stability and toxicity of amyloid fibrils and their
precursors
Titolo progetto
Study of the structure-toxicity relationship of protein oligomers
Ente finanziatore
Durata progetto
Abstract del progetto
EMBO Young Investigator Programme
From 2005 to end of funding (still available to date)
We will investigate the relationship between the structure of protein
oligomers formed by a model proteins at the molecular level and their
biological effect on cultured cells, primary neurons and animal models
Titolo progetto
Structural and function aspects of lipid bilayers in molecular medicine:
involvement of lipid rafts in human pathology
Cassa di Risparmio Firenze
36 months (2008-2010, but estende to one more year)
The project will investigate the role of natural or synthetic llipid surfaces
with different lipid composition and physicochemical features as
modulators of peptide/protein misfolding and aggregation as well as of
amyloid toxicity.
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Integrative approach to the interplay between Aß, oxidative stress, the
proteasome complex and the autophagic pathway. Effect of
neuroprotective molecules
MIUR and Università di Firenze (cofinanziamento PRIN 2008)
24 months (March 2010 – March 2012)
To characterize in vivo in the TgCRND8 mouse model of ß-amyloid
deposition the molecular bases correlating oxidative damage and
autophagy and ubiquitin-proteasome dysfunctions to the gradual
deposition of Aß and to evaluate the effects of neuroprotective drugs.
Design of small molecule therapeutics for treatment of Alzheimer's
disease based on the discovery of innovative drug targets.
European commission (6FP, ADIT) (project LSHB-CT-2005-511977)
5 years (June 2005-November 2010)
To analyze in vivo the expression and phosphorylation state of new
protein targets in selected brain regions of mouse models of Aβ
overproduction using standard immunohistochemical methodologies. To
analyze phenotype of target over-expression in transgenic mice, generate
inside the project, with respect to neurodegenerative and cognitive
parameters
Oleuropein aglycon and hydroxythyrosol as anti-aggregating molecules
potentially exploitable in Alzheimer’s disease prevention and therapy
Ente finanziatore
Durata progetto
Abstract del progetto
Regione Toscana
4 months (November 2010 – October 2012)
The research will study both in vitro and in vivo the anti-aggregating
effect on Abeta42 of oleuropein aglycone and its derivative
hydroxytyrosol, to assess the possible use of these molecules for
prevention and treatment of Alzheimer’s disease.
2. PHARMACOLOGY
E- Non-Pharmacological Therapies
Country: Italy
Contact person: Pio E. Ricci Bitti, M.D. (Dept. of Psychology, University of Bologna)
[email protected]
Date: January 29, 2010
I)
Strategic Issues ( Justification of the importance of the issue )
2.2.1 Brain and brain-related diseases
An increasing number of Parkinson disease (PD) patients are undergoing deep brain stimulation DBS).
It is now important to evaluate not only the benefits concerning the main symptoms related to movement,
but also the psychological (cognitive, affective and behavioural) impact of DBS.
II)
Priority areas
In relation to our experience with PD patients, the most important area to be evaluated by a psychological
point of view is the possible influence of DBS on cognitive processes (such as memory, spatial orientation,
time perception and perspective…).
III)
Impact of the research area
The results of this kind of studies could be used in order to propose specific
psychoeducational interventions to improve the cognitive efficacy of PD patients that underwent DBS
Nome
Contatti
Istituto/Dipartimento
Maurizio Taglialatela, MD PhD – Professor of Pharmacology
Claudio Russo, PhD – Associate Professor of Pharmacoloy
Alfonso Di Costanzo MD – Associate Professor of Neurology
Giovanni Scapagnini MD PhD – Associate Professor of Clinical
Biochemistry
Email: [email protected]
Tel. +39-0874-404894/4851/4891
Dept. of Health Science, University of Molise
Via De Sanctis, 8610 Campobasso - ITALY
Proposta di ricerca
Area di interesse
identificata
Physical exercise and cognitive decline. Epidemiological studies have shown that
physical exercise can delay the occurrence and the progression of dementia, but also to
improve physical, cognitive and psychological performances, with a positive impact on
the quality of life. The mechanisms involved and the identification of potential
biomarkers predictive of a positive response to physical activity are areas of intense
investigation. In our group, we are currently investigating the potential preventive role of
physical exercise programs on cognitive decline in subjects at risk to develop dementia.
Our focus will be on subjects presenting with a subjective memory disturbance or
affected by mild cognitive impairment (MCI).
• Tedeschi G., et al. Brain atrophy and lesion load in a large population of patients with
multiple sclerosis. Neurology. 2005 Jul 26;65(2):280-5.
• Tedeschi G., et al. Correlation between fatigue and brain atrophy and lesion load in multiple
sclerosis patients independent of disability. J Neurol Sci. 2007 Dec 15;263(1-2):15-9.
Finanziamenti ricevuti
Titolo
progetto
Regulation of K+ channels by APP-interacting proteins: functional characterization and
implications for AD-related neurodegeneration
Ente
finanziatore
Regione Campania (2000-2003)
Durata
progetto
3 years
Abstract del
progetto
Recent experimental evidence suggest that K+ channels play a major pathogenetic role in
Alzheimer disease. In the CNS, phosphorylation of voltage-gated K+ channels by tyrosine
kinases (PTK) is a fundamental process regulating their functional activity; its inhibition exerts a
neuroprotective action in several experimental models of neuronal damage. On the other hand,
the expression of several subclasses of K+ channels can reduce cellular tyrosine kinase activity,
thus suggesting the existence of a reciprocal interaction between PTKs/PTPases and K+
channels. Given that the cytosolic domain of β-APP influences, through its adaptor proteins
Dab (disabled), X11 and Fe65, the tyrosine kinase activity of Abl, in the present proposal we
will:
Titolo
E. Evaluate the potential modulation of several classes of K+ channels (Kv1.2, Kv1.3, Kv1.5,
Kv2.1, ERG, KCNQ2+3) by adaptor proteins able to bind to β-APP (X11, Fe65, Dab).
F. Verify the participation of tyrosine kinase activity (and in particular of Abl), in this
modulation.
G. Investigate the potential involvement of these channels in the cellular damage induced by
the hyperexpression of β-APP adaptor proteins.
H. Study the participation of K+ channels and their regulation by oxidative stress in the
neuronal death triggered by various cellular models of AD-related neurodegeneration.
European Community contract N° LSHM-CT-2003-503330/Apopis : Abnormal proteins in the
progetto
pathogenesis of neurodegenerative disorders
Study on tyrosine phosphorylation of APP and its interaction with intracellular adaptors: role in
cell signaling and in the generation of amyloidogenic fragments.
Ente
finanziatore
European Community
Durata
progetto
3 years (2004-2007)
Abstract del
progetto
Within the WP 1 Generation and Turnover, our proposal is aimed to the definition of the role
that posttrasductional modifications of APP, in particular its phosphorylation, may exert on its
pathophysiologycal function. We will investigate both the role of phosphorylated APP/CTFs in
cell signaling, and/or in the generation of amyloidogenic fragments and plaques through the
interaction with intracellular adaptors, as well as the effect that site-specific tyr-phosphorylation
would have on the direct amyloidogenic pathway of APP. These studies, which are focused in a
completely new aspect of APP activity, would lead to the identification of early markers of the
neurodegenerative process which occurs in AD, and to the definition of a potentially innovative
therapeutical approach based on the findings obtained. Our proposal is aimed at:
Aim 1) To study directly in human brain the tyr-phosphorylation of APP and its CTFs and their
possible coupling with intracellular adaptors such as ShcA, and to correlate these data with the
presence of neuropathologycal hallmarks of the disease. The comparative analysis in non-AD
control, AD and/or Down’s syndrome subjects at different ages (from fetal to adult life) will
allow a deeper characterization of the molecular determinants during the progression of the
disease.
Aim 2) To study in transgenic mice carrying APP and PS1 mutations the tyr-phosphorylation
of APP and CTFs, their interaction with ShcA-Grb2 adaptors and the influence that such
interaction could have on plaque formation, astroglial response, neuronal death. The use of a
Tg model will allow at a deeper comprehension of the possible correlation between APP
phosphorylation, its cleavage, signaling activity and progressive presence of the typical
hallmarks of the disease. Moreover, in this model is possible also to determine the effect that
APP and PS1 mutations, which are linked with a human familial phenotype, would cause in the
above mentioned parameters, and, in a second step, would allow the application of targeted
therapeutical pharmacological approaches following also the indications raised from the in vitro
studies (aim 3).
Aim 3) To study in neuronal and/or glial cell cultures the molecular mechanisms which regulate
APP phosphorylation, influence on cleavage by secretases, coupling with intracellular adaptors,
and signaling activity. Using APP mutants, ShcA mutants, and APP KO cells we will define the
influence that phosphorylation and interaction with ShcA would cause in signaling activity, cell
proliferation and death, and in the generation of amyloidogenic fragments. Moreover, the study
of the effect that proliferative and apoptotic stimuli may cause on those parameters and/or with
the study of the kinases involved in the activation of APP cleavage and signaling, this would
also lead to the identification of pharmacological target for a therapeutical intervention.
Nome
Dr. Andrea Stracciari, dr.ssa Maria Guarino, prof. Fabio
Cirignotta
Contatti
Istituto/Dipartimento
Unità Operativa di Neurologia
Azienda Ospedaliero-Universitaria S. Orsola-Malpighi, Bologna
Dipartimento Scienze Neurologiche - Università di Bologna
Proposta di ricerca
Area di interesse identificata
New treatment strategies (non pharmacologic therapeutic
strategies in neurodegenerative diseases)
Finanziamenti ricevuti
Titolo progetto
Cognitive training in Parkinson’s disease
Ente finanziatore
Fondazione del Monte di Bologna e Ravenna
Durata progetto
24 mesi
Abstract del progetto
Background. Patients with Parkinson’s disease (PD) frequently
present cognitive disturbances, which impair their autonomy and
quality of life and are correlated with a significantly increased death
rate.
Objectives. The primary objective is to assess improvements in
quality of life in PD patients undergoing cognitive training.
Secondary end-points will be changes in cognitive capacity,
autonomy, mood, clinical judgement and burden to caregivers.
Methods. This is a prospective randomized, 8-week, single blind, 2arm, multicenter, parallel-group study to evaluate the effect of task
specific cognitive training on quality of life in patients with cognitive
impairment, but without dementia (MMSE> 24), due to idiopathic
PD. Fifty patients per arm will be randomized to detect a 10 point
difference in the PDQ-39 overall score.
Expected results. An improvement in quality of life obtained by
enhancing cognitive functions, namely executive functions, is
expected a the main result, being considered a clinically relevant
outcome.
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Orazio Zanetti
[email protected]
IRCCS Centro San Giovanni di Dio Fatebenefratelli- Brescia
Non invasive brain stimulation an integrated approach to neurorehabilitation in Alzheimer Disease
Alzheimer’s Disease (AD) is a progressive disorder that impacts memory, language and several other
cognitive functions. Given the limited effectiveness of pharmacological treatments, non-pharmacological
interventions in AD have gained attention in recent years, and there are currently many different
approaches under study, ranging from multi-strategy approaches to cognitive training (Cotelli et al 2006).
Despite the potential therapeutic impact of the non-pharmacological approaches, the neural mechanisms
underlying the beneficial effects of behavioral interventions remain largely unknown. Functional
neuroimaging studies have shown that rehabilitation in patients with developmental and acquired
cerebral damage may lead to functional cortical reorganization, a process mediated by activity-dependent
plasticity mechanisms (Warburton et al 1999; Buckner et al 2004). These “plastic” mechanisms may also
play a role in the aging brain and in Alzheimer Disease (Cotelli et al 2006). An increase in the activation of
areas involved in memory, together with the recruitment of new areas during memory intensive tasks, has
been confirmed using neuroimaging techniques in AD patients who underwent behavioral interventions
(Cotelli et al 2008; Manenti et al 2008; Cotelli et al 2010 in press).
Recent studies have reported enhanced performance on specific cognitive tasks in patients with several
types of neurological disease, after receiving non invasive brain stimulation (BS), i.e., repetitive
Transcranial Magnetic Stimulation (rTMS) or transcranial Current Stimulation (tCS) to specific cortical areas
(Miniussi et al., 2008).
Area di interesse identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
New treatment strategies
Nome
Contatti
Istituto/Dipartimento
PATRIZIA BISIACCHI
049 8276587
DIPARTIMENTO DI PSICOLOGIA GENERALE
Proposta di ricerca
Neurodegeneration occurring during normal ageing affects several cognitive processes, particularly
memory, attention, executive functions. Within these functions there are specific aspects that are
particularly susceptible to impairment due to unhealthy ageing, such as working memory, control
mechanisms and sustained attention. These are every day cognitive functions that are a result of a
sequence of complex neurological processes. The bio-signals (i.e. EEG, NIRS etc) recorded from a set of
specific cognitive tasks have important roles in the assessment of these processes. Discovering features in
signals of speech and visual processing, which are modalities that are must less open to subjective
interpretation, would remove the subjective nature of some existing assessments. These modalities
provide signals that can be collected remotely without significant amounts of resources and enable large
research corpora of data, of great clinical significance, to be gathered effortlessly
Specific focus of our researches is on the assessment of cognitive decline in older persons through the
use of multimodal signal processing (i.e. behavioural, EEG, ERPs, fNIRS, Eye movements etc.) .
Current clinical measures of cognitive status and general wellness of the elderly have several limitations.
Most must be administered by trained professionals, the results of these measures can be affected by
many external factors and the ratings can be affected by subjective scoring. Discharging elderly patients
from hospital can often rely on a clinician’s subjective opinion about the patient’s ability to function in
their own home. The possibility to utilize a remote multimodal assessment will bypass such difficulties.
Area di interesse
identificata
-
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Early diagnosis
Imaging techniques and protocols
Standardization of diagnostic criteria and diagnostic
instruments, harmonization and assessment tools
New treatment strategies
Ambient Assisted Living, Home automation, smart
homes and domotics
Cognitive Ergonomics, ICT for elderly/Gerontechnology
ELDERGAMES (Development of High Therapeutic Value IST-based
Games for Monitoring and Improving the Quality of Life of Elderly
People)
PRINCIPAL INVESTIGATOR: PROF L. GAMBERINI
EU FP6
€ 100.373,00
01/09/06 - 28/02/09
Motivating and amusing environments could help elderly
people to preserve their cognitive functions and to improve their
social life. Based on these premises, ELDERGAMES project
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
developed a mixed reality table-top solution on which users have to
exploit and train their cognitive abilities through mind-challenging
games in both collaborative and competitive settings
SENIORCHANNEL (ICT based solutions for the advancement of
social interaction of elderly people)
PRINCIPAL INVESTIGATOR: PROF L. GAMBERINI
EU- AAL/ MIUR
€ 303.000,00
2010-2012
Successful aging requires the involvement of elderly people
in activities such as interacting and sharing their knowledge,
opinions and aspirations with the wider community.
SENIORCHANNEL project aims at supporting these activities
through the development of a low-cost interactive IPTV and TV
studio, through which elderly people will be able to create, interact
and enjoy contents exploiting their creativity and knowledge.
EARLY COGNITIVE AND ELECTROENCEPHALOGRAPHIC MARKERS OF
NORMAL AND PATHOLOGICAL AGEING
PRINCIPAL INVESTIGATOR: PROF P. BISIACCHI
CARIPARO (Cassa di Risparmio di Padova e Rovigo)
€ 45.000,00
January 2009 – December 2011
The main goal of the project is to investigate cognitive and
electrophysiological alterations in healthy old individuals and in
MCI patients, in order to early identify markers of pathological
ageing.
LIFESPAN CHANGES IN ELECTROPHYSIOLOGICAL PATTERNS
ASSOCIATED WITH TEMPORAL DISCRIMINATION
PRINCIPAL INVESTIGATOR: PROF P. BISIACCHI
Bial Foundation
€ 45.000,00
January 2009 – December 2011
The present project is aimed at clarifying the specific contribution
of frontal and parietal brain regions in time processing, and their
development and decline with age by means of new EEG and ERPs
methodologies of signal analyses. Results might further have
clinical application in diagnosis and rehabilitation of Parkinson’s
disease and dementia.
AN INTEGRATED APPROACH TO NEUROCOGNITIVE ASSESSMENT OF
OLD PEOPLE
PRINCIPAL INVESTIGATOR: PROF P. BISIACCHI
FSE (Fondo Sociale Europeo) Regione Veneto
€ 55.372,00
Durata progetto
March 2009 – February 2010
Abstract del progetto
The present project, thanks to the integration of psychological,
medical and bioengineering competencies, was aimed at
developing systems of analysis that optimize the use of ERP/EEG
methodology in the screening of executive functions in old people.
Titolo progetto
IMPLEMENTATION OF NEW EEG SIGNAL ANALYSES FOR THE
DETECTION OF SUBCLINICAL COGNITIVE IMPAIRMENT
PRINCIPAL INVESTIGATOR: PROF P. BISIACCHI
FSE REGIONE VENETO
€ 49.418,18
01/04/10 - 31/03/11
Ente finanziatore
Durata progetto
Abstract del progetto
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
The goal of the project is to create a new integrate approach to
detect subclinical cognitive alterations in many pathologies.
Cross Ages (Inter-generational learning. From diagnostic to impact
evaluation)
PRINCIPAL INVESTIGATOR: PROF R. DE BENI
EU
€ 126.649,00
01/12/07 - 30/11/09
Promoting successful active ageing and social inclusion of older
people in the local communities through the elaboration of an
integrated lifelong learning model based on the intergenerational
exchange.
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
Vania Broccoli
02 26434616
[email protected]
Neuroscience Division
iPS technology offers also an unprecedented opportunity to recapitulate
both normal and pathologic human tissue formation in vitro, thereby
enabling disease investigation and drug development.
PD might greatly benefit from an approach with iPS, since it is completely
missing a valuable in vitro model of this pathology. In fact, up to now,
primary skin fibroblasts and lymphocytes have been the only
human cells available for investigating the pathogenetic mechanisms of
PD. Although, they provided some insights on the disease, their
completely different nature and metabolism prevents gaining information
on the neuronal-specific degeneration processes. To overcome this
limitation, disease affected neuronal cells
would provide a completely new experimental system where to
investigate the molecular roots of PD.
This is even more important in PD, since also the animal models used for
many years for this pathology have shown some important limitations
failing to recapitulate the normal evolution of the pathological
process observed in the patients. In fact, most of the studies have relied
on mouse models of PD, which might be divided in genetic or
symptomatic. Mutant animals for the PD-genes have permitted the
investigation of specific processes associated to neuronal death such as
oxidative stress and mitochondrial dysfunctions, but usually lack key
features of the human disease as for instance selective degeneration of
dopaminergic neurons.
On the other hand, neurotoxins (6-hydroxydopamine, Rotenone, MPTP,
Paraquate) have been widely used to efficiently induce dopaminergic
neurodegeneration in animals, but again, some of the specific
pathological signs of the human disease are not recapitulated as, for
instance, the formation of Lewy bodies, the dynamics of neuronal death
and a comparable development of the neurobehavioral deficits.
For these reasons, the establishment of an iPS model system for PD might
result particular significant.
Indeed, there are some important features, which might enable this
technology to be particular informative in PD research. First of all, some
different procedures have been proved rather efficient in
generating dopaminergic neurons starting from human ES and iPS cells. In
particular, a treatment with FGF8 and Sonic Hedgehog (Shh) acts a strong
enhancer of the in vitro dopaminergic commitment. These
factors could be associated with different in vitro conditions such as coculture systems with stromal cells or SMAD inhibitors that strongly induce
neuronal specification (Barberi et al., 2003; Chambers et al.,
2009). Moreover, the identification of genes whose mutations cause
dopaminergic neuronal loss in a mainly cell-autonomous fashion allows to
generate lines of iPS cells with a known genetic alteration and
compare them to those derived from PD sporadic cases.
Furthermore, we plan to use iPS-derived dopaminergic neurons as a
source of transplantable cells in 6-hydroxidopamine-treated rats and
mice. Neurons will be grafted in either the striatum or the substantia
nigra and their survival, integration and functional activity will be
analyzed. Behavioral studies will assess whether transplanted cells will be
able to rescue the neurological impairment in these animals.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Modeling Parkinson’s disease by iPS technology: generation of human
affected
dopaminergic neurons and gene disease correction by site-specific
integration
ERANET-NEUON- EU
2010-2013
Induced pluripotent stem (iPS) cells derived from somatic cells of patients
represent an innovative tool for in vitro modeling of diseases and may
provide a source for replacement therapies. In Parkinson disease (PD),
the identification of the pathophysiological mechanisms behind this
common neurodegenerative pathology has been particularly hampered
by the lack of genuine in vitro models and animal models faithful to the
human pathology.
Exploiting new generations of viral vectors expressing the reprogramming
(Oct4, Sox2 and Klf4) genes in fibroblasts from skin biopsies of the
patients, we aim to establish faithful pluripotent iPS cell
ines from sporadic and various monogenic forms of PD carrying
mutations in either SNCA, PINK1, Parkin, LRRK2, DJ-1 or UCH-L1. In order
to create an unprecedented in vitro culture system for PD affected human
dopaminergic (DA) neurons, iPS cells will be differentiated in vitro
through the Shh/Fgf8 stepwise induction protocol. Such patient-specific
neurons will provide a system where we will investigate the mechanisms
of genetic PD such as neuronal morphology, Lewy body formation,
mitochondrial dysfunction, cell viability after oxidative and other
stressors, interactions between different disease proteins, as well as
electrophysiological deficits. For those experiments requiring a pure
population of DA neurons, Pitx3-GFP iPS cell lines will be generated by
means of BACtransgenesis enabling specific labelling of nigral DA
neurons. Thus, pure populations of GFP+ dopaminergic neurons will be
utilized for microarray based transcriptome profiling comparing PD versus
control neuronal populations. These results promise to add precious
information on both cellular and molecular pathological mechanisms
leading ultimately to understand PD progression.
Further, the establishment of human PD affected neurons will offer a
remarkable cell platform for future systematic drug screenings. Finally,
we will exploit an innovative strategy for site-specific integration of the
therapeutic gene based on zinc finger nucleases mediated site-specific
integration. This procedure will complement Parkin mutated iPS cells by
targeting the therapeutic gene in a safe and transcriptionally competent
genomic harbour.
Overall, this project has the ambitious task to pioneer new approaches
for iPS cell generation and combining them to develop safe gene
correction strategies. These new procedures represent a big advantage
for a broad functional characterization of iPS-derived dopaminergic
neurons affected by PD. The successful achievements of our goals will
disclose new opportunities for disease modelling, drug-screenings and
cell replacements strategies in PD.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Neural regeneration in the cerebellum: development of cell replacement
strategies for the management of spinocerebellar ataxias.
Gian Giacomo Consalez, Ferdinando Rossi
02 2643 4838
[email protected], [email protected]
Istituto Scientifico San Raffaele, Università degli Studi di Torino
Proposta di ricerca
Optical And Biomolecular Methods For The Functional Investigation Of Neural Circuits In Vivo.
Area di interesse
rigenerazione in modelli di malattia neurodegenerativa
identificata
Finanziamenti ricevuti
Neural regeneration in the cerebellum: development of cell replacement
strategies for the management of spinocerebellar ataxias.
Ataxia UK
Ente finanziatore
2 anni
Durata progetto
The project will use the postnatal and adult mouse cerebellum as a model system to set up cell
replacement protocols to treat degenerative disorders of the central nervous system and, namely,
spinocerebellar ataxias.
Titolo progetto
In this project, we plan to:
1. determine the ability of PC progenitors isolated from the embryonic cerebellum to proliferate in the
host tissue and integrate into the wildtype and mutant cerebellar cortex
2. analyze the spatio-temporal distribution of cues guiding PCs in their migration into the developing and
adult cerebellar cortex
3. manipulate the system to restore developmentally regulated cues guiding PC progenitor homing and
connectivity
The cerebellum is a highly plastic system whose development is completed after birth. Numerous
observations show that exogenous PCs grafted to the cerebellum may integrate productively and establish
proper synaptic connections with their afferents and post-synaptic targets. Several Mendelian cerebellar
disorders, metabolic or neurodegenerative, feature a selective loss of PCs, fully warranting an attempt to
develop cell replacement strategies.
We plan to analyze the ontogenetic mechanisms that direct the integration of PC progenitors in the
cerebellar cortex of wild type and spinocerebellar ataxia type 2 mice, and determine whether they are still
available (or they can be re-activated) in the adult to promote repair. Our strategy for cerebellar
regeneration will be driven by this analysis, in an attempt to exploit existing cues and to enhance those
that are lost in time, eventually favoring the integration of exogenous precursors in the mature
cerebellum.
Protocols suited to promote the anatomical and functional integration of early PC progenitors into the wt
or mutant mouse cerebellum.
Nome
Federico Schena
Contatti
[email protected]
Istituto/Dipartimento
Dipartimento di Neuroscienze-Sezione Scienze Motorie Università di
Verona & Centro di ricerca in Bioingegneria e Scienze Motorie, Rovereto
Proposta di ricerca
Area di interesse
identificata
Physical activity programs for elderly
subjects with different grades of cognitive
impairment.
B.
Finanziamenti ricevuti
Titolo progetto
Walking program for Alzheimer patients.
Ente finanziatore
Comune di Rovereto – Assessorato alle attività sociali
Fondazione Mazzali - Mantova
3 years
Durata progetto
Abstract del progetto
Today, one of the main purposes of institutional long-term care is to offer
well-organized therapy that also enhances the patients’ functional
abilities. New integrated clinical approaches concerning physical exercise
and Alzheimer’s disease (AD) have received attention in the scientific
literature. Apart from individual rehabilitation, in the special care unit,
patients rarely do physical activity alone; this phenomenon has been seen
to cause a decline in the patients’ functional progress.
Motor inactivity and asthenia become evident in the later stages of AD,
including the ability to step, walk, and sit up independently. There is
evidence, however, that specific physical activities can slow down this
inexorable decline. For example, Fiatarone et al. demonstrated that in
very old subjects high intensity resistance training and nutritional
supplementation increased gait velocity, stair climbing power,
spontaneous activity level, cross-sectional thigh area and maximal leg
strength. Moreover, reports in the literature and recent guidelines
suggest that the oldest-old can enhance cardiovascular function,
flexibility, balance and strength through systematic exercise training.
Most researchers in this field agree that also patients with Alzheimer’s
disease can benefit from a targeted exercise program, because exercise
has additional benefits for elderly dementia patients. [8-10] This notion
has been proven in studies on AD patients; which have reported
numerous positive effects of physical exercise and walking programs:
improvement in walking endurance, better urinary continence, enhanced
communication, reduced depression and an increase in activities of daily
life (ADLs).
However in this institutional scenario, caregiver intervention and
interaction with the medical staff and patients are rather difficult. In a
nursing home, however, these important positive relationships were
easier to manage.
The aim of this program is to determinate whether an institution-based
walking program carried out together with caregivers would reduce the
functional, cognitive and physical decline of patients in the later stages of
Alzheimer’s disease. The walking program will be compared with routine
care in a randomized controlled clinical trial. We hypothesized that
patients in the walking program would show improvements in physical
capacity, cognitive performance and biomedical outcomes compared
with those in the control care group.
Nome
Contatti
Istituto/Dipartimento
Prof. Enrico Granieri
[email protected]
[email protected]
[email protected]
Sezione di Neurologia, Dipartimento di Discipline Medico-chirurgiche della
Comunicazione e del Comportamento, Università di Ferrara
Proposta di ricerca
Area di interesse
identificata
E. Non-Pharmacological Therapies
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Promotion of Adapted Motor Activity among patients with Parkinson’s
disease
Fondazione Cassa di Risparmio di Cento,
University of Ferrara
3 years
An adapted motor activity could improve the pattern of body
accelerometer and quality of life of patients with Parkinson’s disease. A
pleasant, playful motor activity associated with music can give a motor
performance improvement in terms of velocity and acceleration of the
gait, stability in posture and better and rapid execution of movements.
Objective of the study is to demonstrate with different technique and
measures whether or not objective differences are detectable in subjects
with Parkinson’s disease after an adapted motor activity based on
emotional involvement through pleasant exercises, such as music and
dance. For this study we developed a triaxial accelerometer that can be
located in three different locations of the body that allow to measure the
average accelerations in the body trunk.
At least 100 patients suffering from Parkinson’s disease will be evaluated.
The adapted motor activity session will be run twice a week, each of
which lasted 60 minutes. They will be tested at entry of the study, after 4
months of continuative motor activity and after 3 months following the
end of the sessions by accelerometer test, disability scale (UPDRS III),
quality of life questionnaire (SF-36), MMSE, tapping test. Measures of
different domains with tests and objective measures with accelerometric
techniques could improve our capability for the clinical monitoring of
patients with movement disorders as Parkinson disease. In the same
contest we aim to demonstrate the efficacy, in clinical terms, of an
adapted motor activity based on emotional involvement through
pleasant exercises with the support of music as a powerful cognitive and
affective stimuli.
(preliminary results of pilot studies has been shown at the Congresses of
Italian Society of Neurology and LIMPE in 2008, 2009 and 2010, at the
Congresses of Neurology in Moskow [Russia] and Belgrade [Serbia] in
2010. Papers are submitted for publication in neurological journals).
Nome
Dott.ssa Mariachiara Sensi
Contatti
[email protected]
Istituto/Dipartimento
U.O.Neurologia ,Dpartimento di Neuroscienze Riabilitazione,
Ospedale S.Anna Ferrara
Proposta di ricerca
Area di interesse
identificata
E. Non-Pharmacological Therapies
Finanziamenti ricevuti
Ente finanziatore
New treatment strategies (non pharmacologic
therapeutic strategies in neurodegenerative diseases) with Deep brain
Stimulation in dystonic and Parkinson Disease patients.
None
Durata progetto
3 years
Abstract del progetto
To evaluate the long term response of Deep Brain Stimulations on motor
and non motor symptoms in Parkinson disease and Dystonic Patients.
The hypothesis is to test axial symptoms, dysartria, dysfagia, pain, both
clinically and with non invasive imaging techniques .
Titolo progetto
2. PHARMACOLOGY
F. Drug Pharmacogenetics
Nome
Alberto Pilotto
Contatti
Phone: +39 0882 410271
Fax:
+39 0882 410271
E-mail: [email protected]
Unità Operativa di Geriatria & Laboratorio di Gerontologia-Geriatria
Dipartimento di Scienze Mediche
IRCCS Casa Sollievo della Sofferenza
Viale Cappuccini, 1 – 71013 San Giovanni Rotondo (FG)
Istituto/Dipartimento
Proposta di ricerca
Sporadic Alzheimer’s disease (AD) is a progressive neurodegenerative disorder occurring predominantly in
older age. The prevalence of AD rise from 20% after 75 years to 30% after 85 years, and about 5% of
people aged 65 years or older have AD. With about 3 to 4 million people affected in the United States and
about 350,000 new cases per year, AD is the most frequent cause of dementia in U.S. and in Western
countries. Among the main cause leading to AD, the deficit of cholinergic system plays a major role.
Accordingly, one of the most common therapy for the symptomatic treatment of AD is the block of
acetylcholinesterase (AChE) by means of acetylcholinesterase inhibitors. The inhibition of AChE increases
the concentration of acetylcholine (ACh) in the synaptic cleft, thus restoring the physiological effects of
ACh within the central nervous system. Recent studies reported a significant benefits of
acetylcholinesterase inhibitors vs placebo on cognitive function, activities of daily living, and behavior.
These improvements, however, are not always detectable in clinical practice. Thus, it has been recently
suggested that the detection of improvements on cognitive function, activities of daily living, and behavior
may be obtained by the concomitant use of drugs acting on other pathogenetic AD mechanism. In
particular the European Medical Agency (EMEA) recommend the use of the AChE inhibitors donepezil and
rivastigmine in mild-to-moderate AD, and the addition of memantine in moderate-to-severe AD.
Memantine block the glutamate receptor, limiting the uncontrolled entrance of Ca2+ ion in the
postsynaptic neuron, thus delaying the Ca2+-excess neurodegeneration. Most studies reported that
interindividual differences in response to these drugs may be due to variability in drug metabolism related
to behavioral, clinical, and genetic factors, mainly hereditary polymorphisms of drug-metabolizing and
drug-transporting enzymes. Final objective of this proposal is to identify the genetic component
underlying the response/non response to the most common drugs currently used in the treatment of AD,
i.e. donepezil, rivastigmine and memantine, throughout the following specific objectives: 1) identification
of significant difference in the distribution of the genotypes of the CYP2D6 gene polymorphisms among
patients responders/non-responders to donepezil treatment; 2) identification of the specific DNA
alteration producing modifications in the CYP2D6 enzyme activity; 3) confirmation of the modification of
the enzyme activity by means of donepezil dosage in vivo; 4) identification of significant difference in the
distribution of the genotypes of the acetylcholinesterase gene polymorphisms among patients
responders/non-responders to rivastigmine treatment; 5) identification of the specific DNA alteration
producing modifications in the acetylcholinesterase-mediated hydrolysis of rivastigmine; 6) confirmation
of the modification of the enzyme activity by means of rivastigmine dosage in vivo; 7) identification of
significant difference in the distribution of the genotypes of the OCT2 gene polymorphisms among
patients responders/non-responders to memantine treatment; 8) identification of the specific DNA
alteration producing modifications in the OCT2 transporter of memantine; 9) confirmation of the
modification of the enzyme activity by means of memantine dosage in vivo. We expected that the
identification of functional polymorphisms in the CYP2D6, AChE and OCT2 genes may influence the clinical
efficacy of donepezil, rivastigmine and memantine in AD patients, and may be useful in identifying
subgroups of AD patients with different clinical response to treatment. Criteria and indicators to verify
results are: 1) the number of patients enrolled in the study in the first 6 months; 2) the number of
genotypes investigated at the end of the 1th year of the project; 3) the number of specific gene alteration
influencing the enzyme activity at the 2nd year of the project; 4) the quantitative data obtained from
plasma drug dosage at the end of the 2nd year of the project. This project is coherent with the ministerial
guidelines for the I.R.C.C.S. and well integrate the clinical practice in neurology.
Area di interesse
identificata
Pharmacogenetics of drugs for Alzheimer’s Disease
Finanziamenti ricevuti
Ente finanziatore
Effect of weight loss on metabolic, functional, cognitive status and
biological markers of longevity in obese frail elderly
Ministero della Salute
Durata progetto
Two years
Abstract del progetto
Obesity, and in particular abdominal obesity causes serious metabolic and
medical complications and impairs quality of life. Moreover, in elderly
persons, obesity can lead to frailty by promoting chronic inflammation
and by exacerbating the decline in strength, endurance, balance and
mobility associated with aging and physical inactivity. Weight loss,
induced by calorie restriction and/or physical exercise, simultaneously
improves multiple metabolic risk factors for cardiovascular disease and
other medical abnormalities associated with obesity, and reduce
morbidity and mortality. However, the appropriate treatment for obesity
in elderly persons is controversial because of the potential harmful
effects of weight loss on bone and muscle mass. It has been reported that
weight loss can also modify biological pathways at the cellular level.
Insulin and of insulin-like growth factor-1 (IGF-1) have been involved in
cellular caloric equilibrium and other mechanisms, i.e. oxidative stress
and inflammation. Thus genetic polymorphisms in genes involved in these
metabolic pathways such as apolipoprotein E (APOE), sterol regulatory
element-binding protein cleavage-activating protein (SCAP), peroxisome
proliferators-activated receptor gamma-2 (PPRγ), α-2B-adrenergic
receptor (ADRA2B), acyl-CoA synthetase 5 (ACSL5) and interleukin-6 (IL-6)
may influence the effect of weight loss on biological mechanisms and
possibly on functional and cognitive status in the frail elderly. It is possible
that weight loss in obese elderly persons can be beneficial by improving
metabolic, functional and cognitive status, or harmful by causing a
decrease in muscle and bone mass. However, the clinical and
physiological effects of weight loss in obese elderly subjects have never
been carefully evaluated. Thus, appropriate treatment for this rapidly
increasing segment of the elderly population remains controversial. The
purpose of this project is to advance our scientific knowledge by
determining the effects of weight loss on metabolic, functional and
cognitive status in obese elderly subjects. Moreover, we will evaluate the
role of weight loss on biological markers of metabolic health and
longevity, i.e. serum level of Insulin, insulin-like growth factor-1 (IGF-1),
transforming growth factor-β (TGF-β), tumor-necrosis factor-α (TNF-α),
interleukin-1 (IL-1) and C-Reactive Protein (CRP), and their relationships
with genetic polymorphisms in APOE, SCAP, PPR-γ, ADRA2B, ACSL5 and IL6 genes. As biomarker of longevity we will also evaluate the accumulation
of mitochondrial (mt) DNA mutation through the analysis of the D310
mononucleotide repeat of mtDNA.
Titolo progetto
Titolo progetto
Smart Home for Elderly People (HOPE)
Ente finanziatore
Ministero della salute/Ministero dell’Università e della Ricerca Scientifica
Durata progetto
Two years
Abstract del progetto
Since Europe’s and worldwide population grows older, the need for care
is growing as well as people spend more money to get it. The solution
Smart Home for Elderly People (HOPE) promises to reduce the need for
carers, improve the life of older people and cut the cost of assistance. The
main objective of the “Hope” project is to produce an Integrated
Computer Technology (ICT) solution that will help the elderly people,
specifically those that suffer with the Alzheimer s disease (AD), achieve a
richer and more independent lifestyle. Dementia causes long and
oppressive suffering to patients and their relatives and imposes
enormous costs on society. About 25 million people suffered from
dementia in recent years. As a 4-fold increase of this number is expected
by 2050, dementia is one main health issue of the next decades. AD
covers 50-70% of all dementia cases, no cure exists, and effective and
reliable early diagnostic techniques are lacking. Early diagnosis and
progress monitoring of AD is a central part of treatment until future drugs
and prevention strategies become available. Elderly people with AD
spend most of their time at home. The “HOPE” solution consists of an
integrated, smart platform that will enable the elderly people with AD to
use innovative technology for a more independent life, easy access to
information, monitor their health, and serve as a source of inspiration for
users as well as for people working with assistive devices. Moreover, it
will enable them to perform by themselves activities they were not able
to do before and which are important for their daily personal life. The
main advantage of the proposed system is that it provides a basis for
integrating services for the elderly population while they are at home.
HOPE is a budgeted solution that will be installed at the elderly people’s
homes, and will provide services for (a) life-long, self organized,
appropriate educational environment and access to information, (b) care
management and health support, (c) self monitoring and decision making.
Optionally, the user will be able to activate software which automatically
establishes the necessary interactive, triple-play connection for receiving
tele-help and tele-assistance from specialized service providers, doctors
or other medical personnel. The proposed application will be intelligent
enough to offer safety in terms of controlling efficiently the home
environment, economy in terms of controlling and decreasing the need
for external help, and convenience in terms of adjusting the operation of
connected sub-systems to achieve the best possible user experience.
2. PHARMACOLOGY
G. Clinical studies
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Prof. Carlo Caltagirone
Via Ardeatina, 306 – 00179 Rome
06 51501409
[email protected]
IRCCS Fondazione Santa Lucia – Laboratory of Clinical and Behavioral
Neurology.
Proposta di ricerca
Area di interesse
identificata
Neurocognition in Alzheimer Disease
Finanziamenti ricevuti
Ente finanziatore
Mortality of elderly using conventional and non conventional
antipsychotic drugs for the behavioural and psychological symptoms of
dementia and for other psychical disorders.
AIFA
Durata progetto
2007-2010
Titolo progetto
Abstract del progetto
Background. Antipsychotic drugs are used to control the
behavioral and psychological disturbances that often occur in
elderly, particularly in those who suffer from dementia. Nonconventional drugs are perceived by clinicians to be better tolerated
than conventional ones and for this reason are more frequently
adopted for frail and elderly patients. However, reports from
ongoing randomized clinical trials on risperidone and olanzapine
(two non conventional antipsychotic drugs) in elderly with dementia,
signaled an increased mortality and incidence of cerebrovascular
events in patients assigned to active drug with respect to placebo.
After this initial report, a formal meta-analysis of fifteen randomized
placebo-controlled trials, (3,353 patients randomized to
nonconventional antipsychotic drugs and 1,757 randomized to
placebo) confirmed the existence of a small increase in mortality in
those patients who had been assigned to active drugs. For this
reason, in European and extra European countries, the health
authorities introduced some limitations to the use of these non
conventional drugs. When a drug treatment for behavioral
disturbances becomes necessary, these limitative measures favor
the use of conventional drugs but it is not known whether
conventional antipsychotics are safer than conventional ones.
Objectives. To assess, in a large population of elderly
residents in different Italian Regions (Piemonte, Emilia,
Marche) and in the metropolitan area of Milan the overall
and cause specific mortality associated with the use of
conventional and non conventional antipsychotic drugs.
Methods. Using the informative systems of the regional health
authorities and of the health district of Milan we will compose a
population cohort formed by all the persons of 60 years or older
who were prescribed an antipsychotic drug for the first time from
January 2001. It is expected that this cohort might be of over 30000
elderly persons. For each included person the last date of existence
in life or the date of death will be retrieved. Mortality will be
evaluated from first assumption of antipsychotic drug by type of
drug used (conventional, non conventional) and by great groups of
causes of death (tumors, cardio vascular diseases and other
causes). Mortality will be separately assessed for elderly users of
antipsychotic and antidementia drugs.
Expected results. This retrospective analysis of survival on an
extraordinarily large cohort will give further evidences about the
potential role of treatment with antipsychotic drugs in survival of
elderly with and without dementia.
3. HUMAN/CLINICAL RESEARCH
A . Human –epidemiology/risk factors
Country: Italy
Contact person: Gabbianelli Rosita or Cinzia Nasuti
School of Pharmacy, University of Camerino,
Date: 29/01/2010
I)
Strategic Issues
The main focus of our research is the development of a progressive neurodegenerative PD rat model. The
research is based upon a multidisciplinary and integrated consortium bringing together field based
researchers (genetic, molecular biology, biochemistry, toxicology and chemistry), from 1 Asian and 7
European countries, and 1 SME (University Medical Center Groningen Netherlands; Institut für Organische
Chemie der RWTH, Germany; Biological Research Center of the Hungarian Academy of Sciences, Hungary;
Instituto de Tecnologia Química e Biológica, Portugal; Departamento de Engenharia Química ISEP, Portugal;
Technological Educational Institute of Athens, Greece; Alagappa University, India; SME, Chema Diagnostica
di Marco Fiore, Italy).
Environmental contaminants accumulated through the food chain can perturb normal physiologic
processes leading to the initiation and progression of the Parkinson’s disease (PD). The consequences
deriving from early life exposure to these environmental factors can influence variability of life span and
the health profile of the elderly in the human population.
PD neuropathology is characterized by the degeneration of the nigrostriatal dopaminergic pathway. It is the
second most common neurodegenerative disorder, affecting 1-2 % of the population over the age of 50. PD
prevalence studies show that the number of individuals with PD over age 50 (4.1-4.6 millions) will double
by 2030 in Western European nations (8.7-9.3 million). Moreover the burden of PD will also shift from more
industrialized nations to developing Asian nations (India, China etc.) where survival of individuals is likely
expected to grow with improving economic conditions and health care.
Environmental factors such as lifestyle, diet, pesticides, metals and solvents, seem to be involved in the
development of neurodegeneration by causing epigenetic modification in people with PD over 50 years old.
Pesticides are one of the most important risk factors of epigenetic changes that affect the development of
PD. Permethrin (PERM) is a pesticide mainly used in agriculture, in healthcare, in industrial and domestic
settings, and in public health services. It is employed in tropical areas to prevent mosquito-borne disease.
Several studies find that pyrethroids persist in house dust in significant concentrations for months after
they are applied for pest control operation and specifically, their metabolites were found in greater than 50
% in the urine of the subjects tested (Riederer et al., 2008). The potential consequences of pesticide
exposure are greater for children than for adults. Children have higher risk of pesticide exposure due to
behaviours such as mouthing items and playing on floors. Nevertheless, the primary exposure pathway of
pyrethroids in humans is thought to be through dietary intake of vegetables, fruits and cereals. It is a
consequence of pesticide residues that has accumulated in soil and groundwater that is re-cycled for
irrigation of crops.
Given the complexity of the many factors to which the populations described in the PD epidemiological
studies have been exposed, the criteria to identify candidate substances as causative factors of PD have to
follow these points: 1) effects on the striatal dopaminergic system (decrease in dopamine level/increase of
its metabolites, decreased in dopamine transporter and enzyme tyrosine hydroxylase activity; 2) damages
on substantia nigra and striatum; 3) mechanistic effects (e.g. on oxidative stress, mitochondrial
dysfunction/complex I inhibition, and alpha-synuclein aggregation). Our previous studies show that
permethrin insecticide, belonging to the pyrethroids family, could be the ideal candidate to satisfy the main
criteria suggested above. Firstly, mitochondria complex I activity was inhibited in striatum cells incubated
with PERM (paper submitted to Neuroscience). Secondly, in vivo studies on pup rats treated with PERM
during brain development (from 6th to 15th day of life) showed an imbalance in the dopaminergic system
(Nasuti et al., 2007). High homovanillic acid levels and reduced dopamine concentrations were measured in
the striatum of 35 day old rats treated. The period of treatment corresponding to neurodevelopment
events (synaptogenesis) in rat brain, occurs in the third trimester of pregnancy in humans. Increased
oxidative stress in striatum (glutathione depletion, increase of protein oxidation), blood cells (decrease of
glutathione peroxidase, increase of lipid peroxidation, unbalance of monocytes and neuthrophils
respiratory burst), and behavioural modifications were measured (Nasuti et al., 2007; Gabbianelli et al.,
2009, a; Gabbianelli et al., 2009, b). Thirdly, markers related to oxidative stress, such as DNA damage,
changes in plasma membrane fluidity and antioxidant enzyme activities were observed in both striatum
and blood cells of adult rats treated with PERM for two months (Nasuti et al.,2003, 2008; Gabbianelli et al.,
2002, 2004; paper submitted to Neuroscience).
Based on assumptions described above we suggest the followed strategic issue: Pathogenic mechanisms,
early diagnosis and prevention of Parkinson’s disease (PD) following pesticide intake.
The research aims to capitalize on knowledge acquired in the field of pesticide effects on
neurodegenerative disease starting from developmental to old age. Studies of the role of epigenetic
mechanisms are a research priority. The studies take into account new approaches for early diagnosis of PD
and strategies able to prevent the development of the neurodegeneration.
Five major objectives to reach:
1) Define the role of the early-life pesticide intake as a causative environmental agent for the development
of PD;
2) Define the correlation between pesticide and epigenetic mechanisms in regulating transcription of
genes;
3) Identify the specific peripheral markers related to early diagnosis of PD;
4) Evaluate the risk following pesticide food intake and translate the outcome to human population;
5) Prevention of PD onset and/or progression by three strategies: reduction of pesticide residues in the
environment through bioremediation, screening of compounds for neuroprotective therapy and
development of diagnostic kits to monitor peripheral markers related to the early PD diagnosis.
Country: Italy
Contact person: Micaela Caserta (CNR)
Date: January 28th, 2010
I)
Strategic Issues
Increase in longevity does not correlate with an increase in disease-free life expectancy. The molecular
mechanisms at the basis of this lack of correlation are still matter of debate and require research
investment.
Cellular senescence has long been used as a cellular model for understanding mechanisms underlying the
ageing process. Compelling evidence obtained in recent years demonstrate that DNA damage is a common
mediator for both replicative senescence, which is triggered by telomere shortening, and premature
cellular senescence induced by various stressors such as oncogenic stress and oxidative stress. Extensive
observations suggest that DNA damage accumulates with age and that this may be due to an increase in
production of reactive oxygen species (ROS) and a decline in DNA repair capacity. Genomic instability is
therefore thought to play a causative role in the ageing process.
Our working hypothesis is based on the possibility that chronic exposure to environmental factors causing
oxidative stress during life, including pollutants but also noxious nutrients such as alcohol, could actually
reduce the ability of relevant genes, implicated in the process of protecting the cell from protein and DNA
damage, to respond properly. If, at the same time, inhibition of programmed cell death (apoptosis) occurs,
cellular damage starts to accumulate.
We will develop experimental strategies to analyse the mechanisms regulating the delicate balance
between rescue and apoptotic pathways in two model systems, human monocytic cells and yeast S.
cerevisiae, with the specific aim to define molecular targets of pharmacological intervention.
II)
Priority areas
Human cells
The major goal is to understand the molecular mechanisms by which environmental insults, causing
oxidative stress in several cell types and tissues, alter the expression of genes involved in counterbalancing
the damaging effects, like those coding for inducible heat shock proteins, detoxifying enzymes, proteins
involved in DNA repair processes as well as factors controlling programmed cell death.
One of the experimental systems that will be employed consist of human monocytic cell lines, that will be
subjected to various stress agents (mutagenic stress, ethanol, heat shock, and oxidative stress) either as
single treatment or as repeated cycles of treatment. The parameters that will be evaluated are:
- mRNA level of selected genes (Hsp70, Hsp90, TRAP1, ERCC1, TDG, MCL1, DAD1);
- chromatin remodelling and histone modifications occurring at their promoters;
- induction of apoptosis/necrosis;
- epigenetic modifications and activity of PARP-1 and SIRT1, two key regulators of the balance between life
and death of stressed cells.
The subsequent step will be to evaluate some of these parameters (those requiring a limited number of
cells) on primary blood monocytic cells.
S. cerevisiae
Saccharomyces cerevisiae has played an important role as a model system to understand the biochemistry
and molecular biology of mammalian cells. The genetic tools available and the short life span have also
made S. cerevisiae a powerful system to study aging. The yeast chronological life span (CLS) is a measure of
the survival of a non-dividing population of cells, and thus can model aging of mammalian non-dividing cells
but also of higher eukaryotic organisms. The parallel description of the pro-aging role of homologs of Akt,
S6 kinase, adenylate cyclase, and Tor in yeast and in higher eukaryotes, suggests that findings in S.
cerevisiae will be valuable to understand human aging and diseases. Moreover, the similarities between
mitochondria and age-dependent mitochondrial damage in yeast and mammalian cells indicate that S.
cerevisiae is a valuable model to study mitochondrial dysfunction and diseases involving this organelle.
Here, we propose to study the relevance of epigenetic modifications due to the yeast sirtuins (Sir2, HST1-4)
at relevant aging loci such as telomeres and ribosomal DNA genes. Analyses will be carried out in condition
of oxidative stress and at different times of chronological life. The results obtained will be then compared
with parallel treatments in human cells were the sirtuin set (seven members in human) could act in a very
similar way.
III)
Impact
The comprehension of the basic mechanisms underlying cell response to environmental cues, alongside
with the increase of knowledge on human biological variation in health and disease, will allow to identify
specific pathways and genes whose activity needs to be strengthened or reduced by pharmacological
intervention.
Nome
Valerio Carelli, MD, PhD
Contatti
[email protected]
Istituto/Dipartimento
Department of Neurological Sciences, University of Bologna
Proposta di ricerca
MELANOPSIN RETINAL GANGLION CELLS AND CIRCADIAN RHYTHMS IN PARKINSON AND
ALZHEIMER DISEASE
Area di interesse
identificata
Biomarkers (chronobiology and neurodegeneration)
Finanziamenti ricevuti
Titolo progetto
MELANOPSIN RETINAL GANGLION CELLS AND CIRCADIAN RHYTHMS IN
PARKINSON AND ALZHEIMER DISEASE
Ente finanziatore
none
Durata progetto
2 years
Abstract del progetto
Background:
Circadian rhythm abnormalities have been described in aging and
neurodegenerative disorders such as Parkinson and Alzheimer disease
(1,2). In particular, the “sundowning” (i.e. the appearance of
psychomotor agitation during the evening-night) described in Alzheimer
patients, has been related to an abnormal circadian rhythm of body
temperature (3). In the last years many retinal abnormalities (mainly
contrast sensitivity but also colour vision defects and others) and the
presence of optic neuropathy have been documented in both Alzheimer
and Parkinson disease (4-6). Moreover, in both diseases bright light
therapy has been reported to be effective (7,8). Nevertheless, no data are
available on the possible relationship between visual dysfunction and
circadian rhythm abnormalities in these disorders. Their possible link
comes from the recent discovery of a new class of photoreceptors, the
melanopsin RGCs (mRGCs) (9). Melanopsin retinal ganglion cells (mRGCs)
are a new class of photoreceptors playing a crucial role in non-image
forming visual functions such as entrainment of circadian rhythms to light
and dark cycle, pupillary light reflex and regulation of synthesis of
melatonin during the night (10). These cells represent about 1% of RGCs
and originate the retinohypothalamic tract (RHT) (11). Furthermore, In
the retina of patients with Parkinson disease a depletion of dopamine
levels has been documented. The amount of this reduction is strictly
influenced by the time of last levodopa assumption (12). It is of note that
dopaminergic amacrine cells interact with mRGCs and regulate
melanopsin gene expression (13-15).
In collaboration with Professor Alfredo Sadun (Doheny Eye Institute, USC,
Los Angeles, California) we recently demonstrated that mRGCs are
partially spared in mitochondrial optic neuropathies, characterized by
selective loss of RGCs (i.e. LHON and DOA). Our study also showed that
mRGCs are lost after age eighty in normal subjects (16,17).
Rationale:
Based on the current knowledge we hypothesize that the abnormalities
of circadian rhythms documented in Alzheimer and Parkinson disease
may be related to the presence of optic neuropathy and involvement of
the mRGCs system.
Aims:
The aims of this project are:
- to verify the presence of rest-activity rhythms by means of actigraphic
recordings in Parkinson and Alzheimer patients
- to verify the presence of optic neuropathy by measurement of nerve
fiber layer thickness, as evaluated with Optical Coherence Tomography
- to verify and quantify the presence of optic neuropathy and the
involvement of mRGCs in post-mortem retinas of Alzheimer and
Parkinson patients
- to verify the presence of dopamine depletion in Parkinson retina
- to correlate the presence of optic neuropathy and circadian rhythm
abnormalities in patients with Alzheimer and Parkinson disease.
References:
1. Wu YH, Swaab DF. Disturbance and strategies for reactivation of
the circadian rhythm system in aging and Alzheimer's disease.
Sleep Med. 2007 Sep;8(6):623-36.
2. Willis GL. Parkinson's disease as a neuroendocrine disorder of
circadian function: dopamine-melatonin imbalance and the visual
system in the genesis and progression of the degenerative
process. Rev Neurosci. 2008;19(4-5):245-316.
3. Volicer L, et al. Sundowning and circadian rhythms in Alzheimer's
disease. Am J Psychiatry. 2001 May;158(5):704-11.
4. Hinton DR, et al. Optic-nerve degeneration in Alzheimer's
disease. N Engl J Med. 1986 Aug 21;315(8):485-7.
5. Sadun AA, Bassi CJ. Optic nerve damage in Alzheimer's disease.
Ophthalmology. 1990 Jan;97(1):9-17.
6. Archibald NK, et al. The retina in Parkinson's disease. Brain. 2009
May;132(Pt 5):1128-45.
7. Paus S, et al. Bright light therapy in Parkinson's disease: a pilot
study. Mov Disord. 2007; 22(10):1495-8.
8. Dowling GA, et al. Effect of morning bright light treatment for
rest-activity disruption in institutionalized patients with severe
Alzheimer's disease. Int Psychogeriatr. 2005; 17(2):221-36.
9. Berson DM, et al. Phototransduction by retinal ganglion cells that
set the circadian clock. Science. 2002 Feb 8;295(5557):1070-3.
10. Hattar S, et al. Melanopsin-containing retinal ganglion cells:
architecture, projections, and intrinsic photosensitivity. Science.
2002 Feb 8;295(5557):1065-70.
11. Hannibal J, Fahrenkrug J. Neuronal input pathways to the brain's
biological clock and their functional significance. Adv Anat
Embryol Cell Biol. 2006;182:1-71.
12. Harnois C, Di Paolo T. Decreased Dopamine in the Retinas of
Patients with Parkinson's Disease. Investigative Ophthalmology &
Visual Science, 1990; 3 1(11)
13. Vugler AA, et al. Dopamine neurones form a discrete plexus with
melanopsin cells in normal and degenerating retina. Exp Neurol.
2007 May; 205(1):26-35.
14. Sakamoto K, et al. Dopamine regulates melanopsin mRNA
expression in intrinsically photosensitive retinal ganglion cells. Eur
J Neurosci. 2005 Dec;22(12):3129-36.
15. Zhang DQ, et al. Wong KY, Sollars PJ, Berson DM, Pickard GE,
McMahon DG. Intraretinal signaling by ganglion cell
photoreceptors to dopaminergic amacrine neurons. Proc Natl
Acad Sci U S A. 2008 Sep 16;105(37):14181-6.
16. La Morgia C et al. Melanopsin-containting retinal ganglion cells
and
circadian
phototransduction
are
spared
by
Neurodegeneration in Mitochondrial Optic Neuropathies.
Neurology March 17, 2009 Suppl 3 A183 (P04.075)
17. La Morgia C et al. Light-induced melatonin suppression in
Mitochondrial Optic Neuropathies. Arvo Annual Meeting, May 3-7
2009 Fort Lauderdale, Florida N. 5663
Nome
Prof. Roberto Lucchini
Contatti
Tel +39030 3996604 Fax +39030 3996080 Email: [email protected]
Istituto/Dipartimento
Dipartimento di Medicina Sperimentale e Applicata, Sezione di Medicina del
Lavoro, Università di Brescia, P.le Spedali Civili 1, 25123 Brescia
Proposta di ricerca
Area di interesse
identificata
Gene-metal interaction as a determinant of neurodegenerative diseases
Finanziamenti ricevuti
1. Titolo progetto
PHIME - Public health impact of long-term, low-level mixed
element exposure in susceptible population strata
Ente finanziatore
EU 6th Frame Program (www.phime.org)
Durata progetto
2006-2011
Abstract del progetto
2.Titolo progetto
PHIME: Gene-environment interaction on manganese induced
parkinsonism are studied in children and elderly resident in areas with
environmental exposure to heavy metals
Epigenomics Studies of Toxic Hazards in Environmental health Research
Ente finanziatore
CARIPLO Foundation, Italy
Durata progetto
2008-2011
Abstract del progetto
Ente finanziatore
This is an interdisciplinary program to conduct in-vitro and human
investigations to evaluate the role of DNA methylation and histone
modifications in mediating the health effects of metals and other
environmental exposures
PROFILO BIOLOGICO E GENETICO DELLA DISFUNZIONE DEI METALLI
NELLA MALATTIA DI ALZHEIMER E NEL ‘MILD COGNITIVE IMPAIRMENT’
Ministero del Lavoro della Salute e delle Politiche Sociali
Durata progetto
2009-2011
Abstract del progetto
Several cross-sectional as well as cohort studies corroborate the concept
of metal implication in Alzheimer’s disease (AD), including clinical
observations suggesting that higher-than normal levels of copper (Squitti
et al, Neurol 2002) and in particular ‘free’ copper – the quantity of serum
copper which is not bound to ceruloplasmin (Squitti et al, Neurol 2005,
2006) – can distinguish AD from healthy controls. Here we tested the
hypothesis that this is the case also for the classification of healthy elderly
subjects vs. Mild Cognitive Impairment (MCI) which represents an
intermediate cognitive state between ‘normalcy’ and full dementia.
3.Titolo progetto
ABSTRACT
Exposure to heavy metals can induce neurodegenerative effects in humans (Zatta et al., 2003). Various
metals like aluminium, manganese, iron, copper, zinc are likely to play important roles in the pathogenesis
of neurodegenerative illnesses such as Alzheimer’s Dementia (AD) and Parkinson’s Disease (PD). In
particular for AD - the most common form of dementia in the elderly -a consensus has been recently
established in the Metal Hypothesis of AD (Bush and Tanzi 2008). This hypothesis suggests that the
interaction of Abeta – the major component of the senile plaques - with specific metals - iron, zinc and
especially Cu - can drive Abeta pathogenicity and AD development and progression. Numerous clinical
studies, including some from our group (Squitti et al, 2002, 2006, 2009) support this hypothesis. In
particular, we have identified in AD patients a serum-level increase in the Cu quantity that does not bind to
ceruloplasmin (free Cu) (Squitti et al, 2005), which correlates with the typical deficits, cerebrospinal
markers (Squitti et al, 2006) and with a worse prognosis of the disease (Squitti et 2009a). The interaction
between environmental exposure with genetic susceptibility is suspected as an important determinant of
these chronic diseases as well (Wright and Baccarelli 2010; Baccarelli e Bollati, 2009). In fact, if on one hand
metabolic dysfunction could have an effect on metal disbalance – especially at the liver lever, being the
liver the controlling organ of metal homeostasis, on the other hand epigenetic factors have been
highlighted in conjunction with oxidative stress mechanism to be triggered by exposure to neurotoxicants
like heavy metals (Zawia et al., 2009). In particular, the effects of metal exposures in early life and
throughout the developmental stages could account for clinical manifestations in older age. Our group has
produced seminal contributions showing that exposure to metals (Wright and Baccarelli 2010; Tarantini
and Baccarelli 2009) modifies global and gene-specific methylation states and that these changes can be
detected in blood DNA. Epidemiological assessment needs accurate reconstruction of long-term and
lifetime exposure, possibly with exposure biomarkers able to reflect long-term cumulative mechanism of
neurotoxicity. Our group has developed various research projects in these areas, covering different aspects
of metal exposure as determinant of AD (Papaleo et al., 2004) and PD (Lucchini et al., 2007; Squitti et al.,
2009b). The understanding of exposure related determinants have important implication in terms of
possible reduction of the epidemiological impacts of neurodegenerative diseases, through preventive
intervention. The line of research we are interested in focuses on an integration of all the aspects –
exposure-related as well as the metabolic ones - of the metal implication in neuro-degenerative disorders.
In particular, we will explore the genetic and epigenetic influence on AD and PD patients exposed to heavy
metals in the highly industrialized province of Brescia, and the hypothesis that gene controlling metal
metabolism – such as the Wilson’s disease ATP7B gene - which controls the free copper levels in the body,
the hemochromatosis HFE gene and Transferrin gene - involved in the regulation of body iron - arbour
susceptibility loci for late-onset AD as well . Moreover, to provide proof-of-concept that a decoppering
approach has a beneficial effect on AD progression, we are currently starting a large randomized controlled
clinical trial based on the concept that metal complexing or ligand agents properly tune the redistribution
of metals in the body. The influence of genes like parkin and PARK9 will also be assessed in relation to
exposure to neurotoxic metals like manganese. Assessment of cumulative exposure (Sinha and Mark, 2005)
to metals will be based on geospatial approach and will be applied to AD and PD patients and age-sex
matched controls from geographical areas with different environmental exposures to heavy metals.
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L, High Prevalence Of Parkinsonian Disorders Associated To Manganese Exposure In The Vicinities Of
Ferroalloy Industries. Am J Ind Med 2007; 50: 11: 788-800
Papaleo B, Alessio, Apostoli P, Battista G, Benedetti F, Lucchini R, Pasqualetti P. Alzheimer disease: study of
occupational risk factors [in Italian]. G Ital Med Lav Ergon 2004;26(4):310-312
Bush AI, Tanzi RE. Therapeutics for Alzheimer's disease based on the metal hypothesis. Neurotherapeutics.
2008 Jul;5(3):421-32. Review
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PM. Elevation of serum copper levels in Alzheimer's disease. Neurology. 2002 Oct 22;59(8):1153-61..
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PM. Excess of serum copper not related to ceruloplasmin in Alzheimer disease. Neurology. 2005 Mar
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P, Calabrese L, Lupoi D, Rossini PM. Excess of nonceruloplasmin serum copper in AD correlates with
MMSE, CSF [beta]-amyloid, and h-tau. Neurology. 2006 Jul 11;67(1):76-82.
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F, Rossini PM. Longitudinal prognostic value of serum "free" copper in patients with Alzheimer disease.
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Nome
Stefano Mattioli
Contatti
Occupational Medicine,
S.Orsola-Malpighi University Hospital
via Pelagio Palagi 9,
I-40138 Bologna, Italy.
Tel: +39-051 636 2761
Fax: +39-051 636 2609
E-mail: [email protected]
Istituto/Dipartimento
Section of Occupational Medicine, Department of Internal
Medicine, Geriatrics and Nephrology, University of Bologna,
Italy
Proposta di ricerca
Area di interesse identificata
Using prevention to reduce the burden of Alzheimer’s
disease (and other ageing diseases)
Finanziamenti ricevuti
Titolo progetto
Occupational exposure to Valsalva manoeuvre and risk of Alzheimer
disease
Ente finanziatore
None
Durata progetto
2-3 Years (6 months for Study design and questionnaire preparation;
at least 1 year for data collection; 6 month for data cleaning, quality
controls and statistical analysis; 6 month for writing the manuscript
and internal revision process)
Abstract del progetto
Introduction
Chronic increased intracranial pressure or exposure to repetitive
intermittent intracranial pressure elevations might contribute to
Alzheimer’s disease (AD) via damage to the choroid plexus resulting
from cumulative effects of repetitive intermittent intracranial
pressure elevations, and leading to a reduction in cerebrospinal
fluid (CSF) production, and hence to diminished CSF clearance of
neurotoxins such as β-amyloid [Wostyn, Can chronic increased
intracranial pressure or exposure to repetitive intermittent
intracranial pressure elevations raise your risk for Alzheimer’s
disease? Med Hypotheses 2004;62:925–30, 2004].
Weightlifters, glassblowers and wind instrument musicians can
generate very high intracranial pressures due to repeated Valsalva
manoeuvres (elevated subglottic pressures), which impair venous
return from the head [Greenfield et al., Transient changes in
cerebral vascular resistance during the Valsalva maneuver in man.
Stroke 1984;15:76–9]. Herein, occupational exposure to Valsalva
manoeuvres might be a risk factor for Alzheimer disease.
Our aim will be the conduction of an etiological study to test the
hypothesis of causal link between occupational history of frequent
Valsalva manoeuvres and the onset of Alzheimer diseases in later
ages.
Methods
We think that a case-control study design would be feasible to our
purpose. Firstly, our hypothesis implies a long latency between
exposure and disease onset. Moreover, regular lifting is a common
task among manual workers. In a previous study conducted in
Bologna, we estimated a lifetime prevalence of manual lifting works
around 32% [Mattioli et al., Physical exertion (lifting) and retinal
detachment among people with myopia. Epidemiology. 2008
Nov;19(6):868-71].
Considering the frequency of Alzheimer disease (10,000 new cases
registered in Emilia-Romagna every year) we can recruit only
incident cases, in order to minimize recall bias.
Selection of cases will be conducted in cooperation with
Neurological Clinic of Bologna; collaboration of relatives will be
asked in order to properly collect information.
Selection of controls (2 for each case) will be based on random
sampling from the national health service registries.
For data collection, we will use a standardized questionnaire
designed for assessment of a series of potential occupational and
non-occupational risk factors, based on those proposed in the
literature. In particular, weight lifting will be assessed both by
analysis of occupational history and by asking specific questions.
Multivariate logistic regression analyses will be conducted to study
the relationship between past occupational exposure to Valsalva
manoeuvres and Alzheimer disease.
Conclusions
Chronic repetitive Valsalva manoeuvres have been proposed has a
possible cause of Alzheimer disease. Weight lifting, a very common
task for manual workers, might play an important role in Alzheimer
diseases aetiology. If confirmed, this hypothesis could have
important impact on the global burden of the disease. Indeed these
findings could have significant implications in prevention, with
important consequences on health and socio-economic fields.
Epidemiological studies are needed to test this hypothesis and to
asses other possible etiologic factors that could be implicated in the
pathologic mechanism [Wostyn, 2004].
Nome
Contatti
Istituto/Dipartimento
Giuseppe Mele (Professore Aggregato di Chimica)
[email protected]
Studio: 0832-297281
Cellulare: 333-9593228
Dipartimento di Ingegneria dell’Innovazione-Università del Salento
Proposta di ricerca
Area di interesse identificata
Costruzione di un “Alzheimer Web-GIS” che consenta di georeferenziare casi di morbo di Alzheimer su aree campione
associando ad ogni caso in esame una “base di dati” contenente
parametri geografici e ambientali allo scopo di capire “se possibile” le
correlazioni tra i parametri (geografici, scientifici, medici, sociali) ed i
casi oggetto di studio.
Nella costruzione del “Web_GIS”, sebbene in ambiti diversi, si potrebbe
trasferire il know-how acquisito nei due progetti di seguito riportati di cui
il proponente è stato Responsabile Scientifico.
Finanziamenti ricevuti
Titolo progetto (1)
Acronimo ECODO-NET
"Development of a model Web based virtual observatory of Acherontas,
Kalamas and Torre Guaceto ecosystems and its application as a mobile
exhibit and permanent environmental kiosk towards public awareness
and sustainable development of coastal ecosystems (EcoDo-net)"
focuses on the improvement of the management, the protection and the
sustainable development of the Greek and Italian coastal ecosystems in
the border area (Acherontas, Kalamas, their intermediary coastal zone
and -Torre Guaceto) through the observation of the ecosystems' quality
and the increase of public awareness about environmental issues.
Programma Interreg IIIA Italia-Grecia 2000-2006
Asse III. Ambiente e Patrimonio Culturale
Misura 3.1 Miglioramento della gestione degli ecosistemi
Ente finanziatore
FESR-Stato-Regione Puglia
Durata progetto (1)
2 anni gennaio /2007-Dicembre/2008
Abstract del progetto (1)
The program "Development of a model Web based virtual observatory of
Acherontas, Kalamas and Torre Guaceto ecosystems and its application as
a mobile exhibit and permanent environmental kiosk towards public
awareness and sustainable development of coastal ecosystems (EcoDonet)" focuses on the improvement of the management, the protection
and the sustainable development of the Greek and Italian coastal
ecosystems in the border area (Acherontas, Kalamas, their intermediary
coastal zone and -Torre Guaceto) through the observation of the
ecosystems' quality and the increase of public awareness about
environmental issues.
Titolo progetto (2)
Linee di intervento per la valorizzazione delle colture agricole locali della
Valle della Cupa
-Web GIS (Geographic Information System) as support for sustainable
development local economy in the fields of agro-industry and fine
chemicals
Ente finanziatore
Consorzio Universitario Interprovinciale Salentino
Consorzio di Comuni della Valle della Cupa
1 anno 2006
Durata progetto(2)
Abstract del progetto (2)
There is nowadays an increasing interest in the development of natural
products and new fine chemicals based on renewable organic materials
using sustainable processes. At the same time innovative developments
associated to main local productions (e.g. olive oil and wine industries) as
well as to the extraction fine chemicals from less important local cultivar
(e.g. mulberry, black-berry, bilberry etc.), are of topical importance
especially in view of a sustainable “green chemistry”.
Emerging technologies for simulating, controlling, and optimizing
complex systems will be considered, crossing a long list of disciplines such
as, botanical, chemical, computational, material science and process
simulation.
In this context we report our studies concerning a planned sampling of
local agri-food and non-food species (traditional and/or minor cultivar),
which could be interesting for this purpose. With this sampling we are
able to trace the real distribution of local cultivations and to map these
on a Geographic Information System (G.I.S.). Then, each sample will be
listed depending on its own biological properties and physical condition
of the selected cultivar.
Nome
Prof. Calogero Caruso MED04
Contatti
Immunopatology (preclinical): Calogero Caruso, email:
[email protected]
Neurology: Roberto Monastero, email: [email protected]
Oral Medicine: Giuseppina Campisi, email: [email protected]
Istituto/Dipartimento
Department of Pathobiology and Medical and Forensic Biotechnologies
Proposta di ricerca
Area di interesse identificata
Experimental (preclinical) research
The preclinical group (Immunopathology Unit) is evaluating the
role of inflammaging in the expression/progression of the disease. So far
the alterations of the inflammatory response and the pro-inflammatory
profile of aged people have been studied as risk factors for AD
development. This aspect has to be further characterized in terms of the
comparison of the genes and proteins involved in determining the proinflammatory profile in healthy old and very old people and in demented
(both Alzheimer and non Alzheimer). On the other hand, persistent
peripheral multibacteria infection such as periodontitis (PD), associated
with gram-anaerobic bacteria capable of exhibiting localized and systemic
infections in the host, has been proposed as possible aggravating cofactor
in subjects with vascular disease. In PD the balance between bacteria and
host response is altered, resulting in a production of high levels of proinflammatory mediators and low levels of anti-inflammatory ones. Data
regarding the PD association with neurodegenerative disorders are
lacking; however, it seems plausible that, during PD, cytokines increase in
AD brain due to stimulation of trigeminal nerve fibres, or directly from
bacterial products. So, the Immunopathology group in association with
the Oral Medicine group is going to gain insight into this relationship,
using biological samples collected from the Neurological group.
Many studies have reported different alterations of the immune
system in AD, and the involvement of the acquired branches of the
immune system. To investigate the systemic signs of immune responses
in AD, the Immunopathology group is also analyzing the expression of
activation markers on peripheral blood mononuclear cells from AD
patients and age-matched healthy controls activated in vitro by
recombinant amyloid peptide (rAβ42) and their cytokine production. This
kind of study is also useful to obtain biomarkers of AD for monitoring the
effectiveness of therapeutic interventions.
Clinical research
Risk factors for Mild Cognitive Impairment (MCI) - which may precede
the clinical detection of AD and dementia - and predictors for the
conversion from MCI to dementia are currently evaluated by the
Neurological group. For this purpose, hospital-based data and crosssectional as well as prospective data from a population-based cohort are
used. In particular, modifiable (non-genetic) and non-modifiable (genetic)
risk factors are under evaluation.
• Concerning modifiable risk factors, three etiologically-driven
hypotheses are currently carried out:
1. The role of modifiable vascular risk factors (eg, diabetes,
hypertension, dyslipidemia, obesity), alone or clustered, in the
concept of metabolic syndrome are under investigation;
2. The role of behavioural symptoms, particularly depression, in
determining the risk of MCI and its progression to dementia/AD
are under evaluation;
3. The effects of oxidative stress and vitamin E on the development
of MCI and dementia/AD, and their effect in the progression from
MCI to dementia/AD are in assessment.
• Concerning non-modifiable risk factors, we are going to evaluate
the role of recently proposed and novel putative genes involved
in AD. We hypothesize to include a large sample sample of AD
and control subjects (case:control of 1,000:1,000); accordingly,
this study would benefit from a valid statistical power in
detecting novel potential genetic markers for AD. These genes
will be evaluated alone, in clusters and in conjunction with the
APOE4 allele. This study will be performed in association with
Immunopathology Unit. Drugs currently in use to treat AD are
effective only in 20% of patients; their therapeutic effect is
predominantly under genetic control. Thus, these data would
encourage new research in pharmacogenomics.
Finanziamenti ricevuti Principal Investigator: Dr. Roberto Monastero ; Amount of fund: 590,000 euros
Titolo progetto
Ente finanziatore
Epidemiology and risk factors for Mild Cognitive Impairment, Alzheimer's
disease and dementia. The Zabut Aging Project: 5-year follow-up study on
a rural Sicilian population
Ministero della Salute, Italia
Durata progetto
3 years
Abstract del progetto
We would extend recent research on risk factors for Mild Cognitive
Impairment (MCI) and Alzheimer's disease (AD) and the progression of
MCI to dementia in the elderly with a special focus regarding modifiable
(non-genetic) and non-modifiable (genetic) risk factors. Baseline (n=1930
subjects) and 5-year population-based data (we are expecting to enrol
approximately 1500 subjects) will, therefore, be used. Firstly, the role of
vascular, modifiable vascular risk factors, alone or clustered in the
concept of metabolic syndrome would be investigated. Secondly, the role
of behavioural symptoms, particularly depression, in determining the risk
of MCI and its progression to dementia/AD would be evaluated. Thirdly,
the effects of oxidative stress and vitamin E on the development of MCI
and dementia/AD, and their effect in the progression from MCI to
dementia/AD would be assessed. Lastly, this project would evaluate the
role of recently proposed and novel putative genes involved in AD.
Roberto Monastero, MD, PhD
Lab of Epidemiology and Psychology of Aging and Dementia
Dept of Clinical Neuroscience
University of Palermo
Via La Loggia 1
90129 - Palermo, Italy
Tel. +39-091-6555185
Fax. +39-091-6555113
email. [email protected]
[email protected]
Type: Progetto Giovani Ricercatori Ministero Salute, 2007
Title: Epidemiology and risk factors for Mild Cognitive Impairment, Alzheimer's disease and dementia. The
Zabut Aging Project: 5-year follow-up study on a rural Sicilian population
Principal Investigator: Dr. Roberto Monastero
Amount of fund: 590,000 euros
Lenght: 3-year project
Abstract
We would extend recent research on risk factors for Mild Cognitive Impairment (MCI) and Alzheimer's
disease (AD) and the progression of MCI to dementia in the elderly with a special focus regarding
modifiable (non-genetic) and non-modifiable (genetic) risk factors. Baseline (n=1930 subjects) and 5-year
population-based data (we are expecting to enrol approximately 1500 subjects) will, therefore, be used.
Firstly, the role of modifiable vascular risk factors, alone or clustered, in the concept of metabolic syndrome
would be investigated. Secondly, the role of behavioural symptoms, particularly depression, in determining
the risk of MCI and its progression to dementia/AD would be evaluated. Thirdly, the effects of oxidative
stress and vitamin E on the development of MCI and dementia/AD, and their effect in the progression from
MCI to dementia/AD would be assessed. Lastly, this project would evaluate the role of recently proposed
and novel putative genes involved in AD. Due to the large, hypothesized sample of AD and control subjects
(about 1,000 for each group), this study would benefit from a valid statistical power in detecting novel
potential genetic markers for AD. These genes would be evaluated alone, in clusters and in conjunction
with the APOE4 allele.
Nome
Emilio Di Maria, MD PhD
Contatti
[email protected]; +39 0105634368
Istituto/Dipartimento
Dept. of Neuroscience, Ophthalmology and Genetics - University of
Genova
and Division of Medical Genetics, Galliera Hospital, Genova, Italy
Proposta di ricerca
Area di interesse
identificata
Problem statement: The clinical manifestations of Alzheimer’s disease (AD)
often include behavioural disturbances, such as apathy and mood disorders,
as well as psychotic symptoms (delusions and hallucinations). These features,
which significantly affect the clinical outcome, are globally referred to as
behavioural and psychological symptoms in dementia (BPSD). BPSD are
recognised to cause a poorer course of disease, contributing to increase the
social, pharmacological, and managing costs of AD. Family studies have
reinforced the hypothesis that the occurrence of psychosis in AD patients is
partially determined by genetic factors.
The present proposal is in accordance to the priorities listed in the letter of
intent for the joint initiative, namely:
Genetic susceptibility to Alzheimer’s disease
Biomarkers
Studying early onset forms of Alzheimer’s disease to follow-up disease
progression.
Objective: to identify genetic risk factors responsible for the occurrence of
behavioural and neuropsychiatric symptoms in patients with Alzheimer’s
disease. We intend to pursue this objective by using both main approaches: i)
genome-wide association studies; ii) candidate gene approach, in that the
analysis will be performed only on genetic markers which were demonstrated
to be associated with neurodegeneration or psychiatric disorders.
To accomplish this plan, a network of Italian clinical centres and laboratory of
genetics with distinguished expertise in the field has already been built.
Preliminary results: A large cohort of patients with AD has been recruited
through a multicentre effort. Patients were assessed at their admittance with
a complete sociodemographic and clinical data collection. The study protocol
was approved by the Institutional Review Boards. Patients enrolment is still
ongoing. By applying the candidate gene approach outlined above, we have
already demonstrated that the DAOA gene is a genetic risk factor for the
occurrence of psychotic symptoms in AD [Di Maria et al, J Alzh Dis 2009].
Relevance: The identification of genetic factors influencing the occurrence of
BPSD in AD will shed new light on the disease pathophysiology and allow a
better prediction of the illness. On the basis of new lines of evidence, earlier
and tailored treatments could be appropriately administered.
Finanziamenti ricevuti
Note: the Italian network on behavioural and psychological symptoms in dementia, quoted in the proposal
section, has not been formally assembled. Therefore, the funded project granted to other
researchers involved could not be reported.
Titolo progetto
MacAge – MCI-AD Conversion: Analysis of Genetic Effects. A cohort
Ente finanziatore
association study on the role of tau gene haplotypes and ApoE in
determining the evolution from mild cognitive impairment to Alzheimer’s
disease.
CARIGE Foundation, Italy
Durata progetto
1 year
Abstract del progetto
Background: The association of microtubule-associated tau protein gene
(MAPT) with Alzheimer’s disease (AD) had been controversial, but recent
literature demonstrated that the MAPT H1c haplotype is a genetic risk
factor for AD [Myers AJ, 2005]. H1c haplotype is associated with
increased tau expression and predisposes, in late-onset AD, to the
unbalance between the 4-repeat- and the 3-repeat-containing isoforms
(4rep/3rep), thus leading to tau dysfunction and deposition [Myers AJ,
2007]. A distortion of the 4rep/3rep expression pattern was also found in
cholinergic neurons from mild cognitive impairment (MCI), a clinical
condition which is prodromal to AD [Ginsberg SD, 2006]. Reducing tau
levels in mice protects against amyloid-β-induced neuronal dysfunction
[Roberson ED, 2007].
Preliminary results: We previously demonstrated that the tau H1
haplotype is a genetic risk factor for corticobasal degeneration, a sporadic
tauopathy [Di Maria E, 2000]. More recently, a longitudinal study (PI:
Prof. M. Tabaton) characterised individuals with amnestic MCI (aMCI), in
order to find biological markers associated with the risk of AD [Assini A,
2004; Odetti P, 2005]. To date, 224 individuals with aMCI were recruited.
We foresee to collect within the first phase of the project a cohort of at
least 240 cases: all followed-up for 1 year, about ⅓ for ≥2 years.
Objective: To evaluate the hypotheses that the evolution from MCI to AD
is influenced by the tau locus, and that its effect is mediated by a
qualitative and quantitative unbalance in the expression of the tau gene
isoforms. The specific aims will be accomplished according to the
following experimental plan (total duration: 1 year).
- Patients evaluation: i) patients follow-up, blood sampling (according
to the current protocol for the longitudinal study); ii) DNA, plasma,
serum, cell lines banking (Galliera Genetic Bank).
- Cohort association study: i) the tau tagging polymorphisms will be
genotyped in the case series; ii) the association study will investigate
the effect of the tau haplotypes on the MCI-AD transition, taking into
account ApoE genotypes, as well as age, age at onset, gender and
possible interacting variables.
- Gene expression study: expression level of the tau isoforms will be
investigated in lymphoblasts from patients belonging to the two
subsets (MCI and MCI-AD converted) and carrying different tau
genotypes.
Relevance and deliverables: The identification of genetic factors
influencing the evolution from MCI to AD will shed new light on the
disease pathophysiology and allow a better prediction of the disease
progression. On the basis of new lines of evidence, earlier and tailored
therapeutic approaches can be envisaged.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Prof. Carlo Caltagirone
Via Aretina, 306 – 00179 Rome
05 51501409
[email protected]
IRCCS Fondazione Santa Lucia – Laboratory of Clinical and Behavioral
Neurology.
Proposta di ricerca
Area di interesse identificata
Preclinical Diagnosis
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Markers to diagnose neurodegenerative diseases and of new targets to
test the pharmacological treatments.
Italian Ministry of Health
Durata progetto
2006-2008
Abstract del progetto
The present investigation involved 200 individual with AD, whose
cognitive functioning was monitored along a 18-months period by means
of administration of the Mini Mental State Examination. The results
document, on one side, a progressive decline of cognitive performance
and, on the other side, a significant relationship between the occurrence
of some specific genome variants and both the severity and the earlier
onset of disease. In particular, the presence of i) allele e4 of ApoE
polymorphism, of ii) allele T of the polymorphism of the serotonin
receptor 5HT2A (102T/C), of iii) some variants of GST genes as well as of
the iv) polymorphism -137C/C of the promoter region for IL-18,
significantly influence both the onset and the clinical disease course.
Moreover, in line with the above data, the analysis of the cellular
components of innate immunity confirmed that the AD patients’ immune
system cells are more likely to produce phlogistic factors such as IL-6 and
IL-18.
Furthermore, it has also been showed that the previously reported
decrease of proteinic level of ADAM10 within the AD patients’ platelets, is
not owed to reduced levels of ADAM10 mRNA but, rather, to others
events that involve the translation processes or the proteolysis of the
molecule itself.
Finally, groundbreaking in vivo and in vitro animals studies indicate that
the Tat-Pro ADAM peptide can produce alterations of the molecular
composition of the glutamatergic synapses, affecting the synaptic
functioning.
In conclusion, the present study adds some valuable information about
the role played by some disease mediators, such as early inflammatory
episodes and synaptic changes, thus, helping to identify new molecular
cascades and cellular processes involved in the pathogenesis of
neurodegeneration and, likely, to be used as markers of the disease.
Indeed, by means of project here presented, we were allowed to
implement an interdisciplinary work to explore the physio-pathogenetic
pathways in AD, involving a wide number of individuals with AD that
belong to several Italian centres. As a matter of fact, in the view of the
multidisciplinary network, relying upon the synergistic coordination
between basic and clinical researchers, the involvement of the most
representative ITINAD units allowed us to reach results that significantly
advance our understanding of AD pathogenesis. Moreover, the same
results might represent relevant clues to individuate an advanced
strategy in order both to early diagnose AD and to formulate an accurate
prognosis.
Nome
Manservigi Roberto
Contatti
Tel. +39 0532 455401/455412
Fax +39 0532 974470
Mail: [email protected]
Istituto/Dipartimento
Department of Experimental and Diagnostic Medicine
Proposta di ricerca
Area di interesse identificata
Neurodegenerative Diseases
Finanziamenti ricevuti
Titolo progetto
Search of herpetic proteins involved in the pathogenesis of Alzheimer.
Ente finanziatore
MiUR
Durata progetto
2006-2008 (two years)
Abstract del progetto
The overall objective of this proposal is to study the correlation between
HSV-1 infection and Alzheimer’s disease (AD) and to understand those
HSV-1 functions that are involved in the pathogenesis of AD.
:
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Search of herpetic proteins involved in the pathogenesis of Alzheimer's
disease for the development of new therapeutic strategies
MiUR
Durata progetto
2008-2010 (two years)
Abstract del progetto
Despite the very numerous studies on Alzheimer's disease (AD), especially
on amyloid plaques and neurofibrillary tangles, little information has been
obtained thus on the cause of the disease. Moreover, different
experimental evidences define a role for Herpes simplex virus type 1 (HSV1) infection as a risk factor for AD development. The goal of this project is
to study and characterize the different interactions between viral proteins
and the disease development, in order to obtain a better understanding of
AD origins and, above all, to develop innovative therapies.
3. HUMAN/CLINICAL RESEARCH
B. Epidemiology genetics
Nome
Contatti
Istituto/Dipartimento
Luisa Benussi, Giuliano Binetti
0339-030-3501725
[email protected]
IRCCS “Centro S. Giovanni di Dio-Fatebenefratelli” NeuroBioGen LabMemory Clinic, Brescia Italy
Proposta di ricerca
Alzheimer disease (AD) is a genetically complex and heterogeneous disorder. Less than 10% of AD patients
are familial with an autosomal dominant inheritance of the disease (FAD). In autosomal dominant AD
families, rare mutations in the amyloid precursor protein gene, and the presenilin 1 and 2 genes have been
identified. In sporadic patients the disease is expected to result from the interaction of multiple
susceptibility genes and unknown environmental factors. So far, the e4 allele of the apolipoprotein E
(APOE) gene has been the only consistently replicated genetic risk factor for late-onset AD. Through
genome wide association studies (GWAS), a novel opportunity of identifying dementia risk genes and
associated disease pathways has emerged with several potential AD risk genes already reported. Further
validation and replication in other patients cohort combined with functional characterization of these
genes could lead to a better understanding of the pathophysiology underlying dementia and open new
therapeutic possibilities for AD. In the last years our group has contributed to: 1) the identification of
genetic factors that influence the disease risk and the pharmacological response in AD patients (Nicosia et
al., Dement Geriatr Cogn Disord. 2001;Ventriglia et al, Mol Psychiatry 2002; Scassellati et al, Neurosci Lett
2004; Ventriglia et al, Aging Clin Exp Res 2005; Emanuele et al, NBA 2009; Di Maria et al, JAD 2010; Santoro
et al, CNS Drugs 2010); 2) the identification of mutations that are associated with familial forms of AD
(Finckh et al, Am J Hum Genet 2000; Finckh et al, Neurology 2000; Binetti et al, Neurosci Lett 2003;Binetti
et al, Ann Neurol 2003; Signorini et al, Curr Alzheimer Res 2004; Alberici et al, Eur J Neurol 2007;Ghidoni et
al, JAD 2009); 3) the assessment of areas of intervention for genetic counseling of dementia by a crosscultural comparison between Italians and Americans (Binetti et al., PEC 2006).
The specific objectives of this proposal are:
1) Identification of genetic factors that influence the disease susceptibility and the pharmacological
response in AD patients (genome wide association studies);
2) Identification of new mutations and novel AD-associated loci (genetic screening; linkage analysis;
quantitative trait - proteomic profiles- linkage analysis);
3) Evaluating the psychosocial impact of genetic testing in families with hereditary forms of AD.
To carry out this project our group will collaborate with: Dr Roberta Ghidoni (Proteomics Unit, IRCCS Fatebenefratelli, Brescia) on genetic and proteomic studies; Dr F Tagliavini (IRCCS Fondazione Istituto Carlo
Besta, Milano), Dr D Galimberti and Dr E Scarpini (University of Milan) on genetics and genetic counseling;
Prof C Franceschi (Università di Bologna), Dr G Forloni (Istituto Mario Negri, Milano) on genetics and
pharmacogenetics.
Our Group, along with the mentioned National collaborators, can offer to the proposed project the
following resources:
1) Clinical expertise in Alzheimer disease; 2) A large collection of biological samples from sporadic and
familial AD patients (more than 1000 index patients) and a similar number of age-matched controls; 3) A
large collection of families with autosomal dominant inheritance trait for AD (more than 90 families); 4)
Genetic testing as well as a genetic counselling program for families with hereditary forms of dementia.
Area di interesse identificata Genetic susceptibility to Alzheimer’s disease and genome wide
association studies
Finanziamenti ricevuti
Titolo progetto
Project1: Validation of genetic and biochemical markers for early
diagnosis of AD and the prediction of conversion of MCI into AD, and
Ente finanziatore
Durata progetto
Abstract del progetto
design of a multivariate molecular protocol having high diagnostic and
prognostic accuracy.
Project2: Marker biologici di neurodegenerazione utilizzabili nella pratica
clinica ai fini della diagnosi precoce e differenziale, della individuazione
delle forme con risposta ottimale alle attuali terapie
Project3:GenEtica- Profili bioetici e biogiuridici della genetica tra ricerca
sperimentale, consulenza e prospettive terapeutiche.
Project1:Italian Ministry of Health Progr Strategico PS39, prog 1
Project2:Italian Ministry of Health Progr Strategico conv.71 prog 6
Project3: Italian Ministry of University and Research, FIRB2006
Project1:01-01-2009/31-12-2010
Project2:7-01-2008/7-04-2010
Project3:15-10-2008/15/10/2011
Project 1 and Project 2 aims:
1) The development and validation of a multi-factorial protocol that
integrates molecular, imaging, neurophysiological, neuropsychological
and behavioral data for early diagnosis of AD, in particular during the
preclinical phase and the prodromal stage of "mild cognitive impairment"
when the symptoms are not severe enough to fulfill the current
diagnostic criteria for AD.
2) Validation of this new diagnostic protocol within a regional public
health network, and evaluation of the health care, organization and
economic implications of its transfer to the National Health Service.
Project3 aims:
1) to evaluate the psychosocial impact of genetic testing in families with
hereditary forms of dementia; 2) to evaluate the efficacy of the pre-test
educational session and the information comprehension during genetic
counselling
3) analysis of the ethical issues arising from a genetic counselling service
for dementia
Nome
Contatti
Istituto/Dipartimento
Maria Del Zompo
Phone: +39 070 6092438
Fax: +39 070 653584
Email: [email protected]
Department of Neurosciences “B.B. Brodie”, University of Cagliari,
Cagliari, Italy
Proposta di ricerca
Area di interesse identificata
-
Genetic susceptibility to Alzheimer’s disease and genome
wide association studies
- Biobanking (blood sample, CSF, brain repository…)
(In the Biobank of the Section of Clinical Pharmacolgy of the Department
of Neuroscience, University of Cagliary, are cryopreserved the
lymphoblastoid cell lines of 147 patients with Alzheimer's disease and 116
controls. AD patients of proven Sardinian ancestry (parents, grandparents
and great-grandparents) were diagnosed according to DSM-IV, and
National Institute of Neurologic and Communicative Disorders and
Stroke–AD and Related Disorders Association (NINCDS-ADRDA) criteria
for possible or probable AD (McKhann et al, 1984 ). Cognitive screening
was performed by means of Mini-Mental State Examination (MMSE)
(Folstein et al, 1975). All control subjects were cognitively intact
volunteers with an MMSE score ≥ 26 and with negative family history for
dementia.)
3. HUMAN/CLINICAL RESEARCH
C. Biomarkers and early diagnosis and imaging
Nome
Contatti
Istituto/Dipartimento
Proposta di ricerca
Area di interesse
identificata
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Finanziamenti ricevuti
Titolo progetto
Patrizia Mecocci
[email protected]
+39 075 5783270
Sezione di Gerontologia e Geriatria
Dipartimento di Medicina Clinica e Sperimentale
Università degli Studi di Perugia
1)Studies on plasma biomarkers of oxidative/nitrosative stress, inflammation,
lipid and glucose metabolism, in older adults, both in population based and
clinical based longitudinal studies. Samples (already collected or to be
collected) derived from Italy and other European countries, with which we are
collaborating, to develop easily available biomarkers that can be used in the
elderly to: a) study age-related changes in metabolism, also in relation to
lifestyle (i.e. diet, physical activity); b) obtain diagnostic and prognostic tools,
which can be used in different age-related disorders (cognitive
decline/dementia; multimorbidity; disability). The main goal is to obtain noninvasive tools that can be used in age-related disorders for i) investigating
pathogenetic mechanisms; ii) diagnosis, prognosis and monitoring of
preventative and therapeutic interventions.
Age-related cognitive decline and dementia are one of our main research areas,
and our studies integrated plasma biomarkers and neuroimaging data.
2) Studie on cellular and animal models of aging, using markers of
oxidative/nitrosative stress and mitochondrial structure and activity to evaluate
the effects of novel therapeutics (chemically modified antioxidants, vitamins,
nutriceuticals) that can be used in humans to prevent age-related cognitive
disorders.
ZINCAGE Contract n° 506850
European Union in the 6th Framework Program (FP6): "Food Quality and
Safety", Priority 5, Thematic Area T6
3 years
This is an epidemiological study on the influence of diet and lifestyle on healthy
ageing, aimed at preventing adult degenerative disease, particularly focusing on
cardiovascular and neurodegenerative diseases and also addressing
malnutrition of the elderly.
InnoMed (Innovative Medicines for Europe) Project. Contract n° 518170.
European Union in the 6th Framework Program (FP6): Life Science Health,
Priority 5.
3 years
The project, led by the European Federation of Pharmaceutical Industry and
Associations (EFPIA), addresses the complex issues associated with the future of
biomedical research in the EU,and addresses ways of achieving accelerated
development of, safe and more effective medicines, aiming to revitalize the
European biopharmaceutical research environment. The project is mainly
devoted to study cognitive impairment in the elderly and Alzheimer’s disease.
K4Care Project (Knowledge-Based Homecare eServices for an Ageing Europe)
Ente finanziatore
Durata progetto
Abstract del progetto
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
Finanziamenti ricevuti
Titolo progetto
Contract n° 026968
European Union in the 6th Framework Program (FP6): Information, Society,
Technology, Priority 2
3 years
The main objective of the K4CARE project is to improve the capabilities of the
new EU society to manage and respond to the increasing number of senior
population requiring a personalized healthcare assistance. The project will
capture and integrate the information, skills, expertises, and experiences of
specialised centres and professionals of several old and new EU countries, and
will incorporate them in an intelligent web platform in order to give service to
health professionals, patients, and citizens in general. Chronic conditions, such
as dementia, are the main focus of the project.
PRIN Oxidative and nitrosative damage of mitochondrial proteins in models of
cell senescence and in subjects with Alzheimer's disease: a pre-clinical study on
natural forms and new synthetic analogues of vitamin E
MIUR
2 years
Mitochondria are key targets of oxidative stress with relevance in cell
senescence (mitochondrial theory of aging) and possibly in the pathogenetic
mechanisms of age-related diseases such as Mild Cognitive Impairment
(MCI)and Alzheimer's disease (AD). In this context emerging aspects deal with
the study of post-transductional modifications (PTMs) in proteins caused by
reactive oxygen or nitrogen species (ROS and RNS, respectively). The aim of this
project is to evaluate PTMs in mitochondrial proteins in "in vitro" models of
cellular senescence as well as in peripheral cells from subjects with MCI and AD
also evaluating the role as protecting agents of natural forms and newly
synthesized analogues of vitamin E.
Ente finanziatore
Young researchers 2007 Epidemiology and risk factors for Mild Cognitive
Impairment, Alzheimer’s disease and dementia. The Zabut Aging Project:
5-year follow-up study on a rural Sicilian population.
Ministero della Salute
Durata progetto
3 years
Abstract del progetto
The study will evaluate the risk factors for MCI and AD and the
progression of MCI to dementia in the elderly with a special focus
regarding modifiable (non-genetic) and non-modifiable (genetic) risk
factors. Baseline and 5-year population-based data will, therefore, be
used. Main topics are i) the role of modifiable vascular risk factors; ii) the
role of behavioural symptoms, particularly depression, in determining the
risk of MCI and its progression to dementia/AD; iii) the effects of
oxidative stress and vitamin E on the development of MCI and
dementia/AD; iv) the role of recently proposed and novel putative genes
involved in would encourage new research in pharmacogenomics.
Nome
Prof. Salvatore Monaco
Contatti
[email protected]
045-8124285/4461
Neuropatologia/Scienze Neurologiche e della Visione
Istituto/Dipartimento
Proposta di ricerca
Dementia is a clinical syndrome characterised by a progressive loss of cognitive abilities and executive
functions, with a significant compromise of patient’s social, relational and working functions. Recent data
suggest that pathological and biochemical deteriorations take place years before the clinical emergence of
the different dementia conditions. The search for novel markers with a high diagnostic sensitivity is
therefore one of the major and high-priority objective of dementia research. There are several
methodologies that are presently used or under development, including proteomics which has been
shown as one of the most promising and useful for the identification of novel markers.
Aim of the present program is therefore to identify clinical and biological markers of dementia according
to the following experimental strategy:
A). To study mild cognitive impairment and Alzheimer’s disease patients by assessing a specific clinical,
neuropsychological, genetic and biochemical profile at the individual level.
B). To identify novel genotypic markers associated to the specific clinical profile from all individual
patients.
C). To identify novel CSF and plasmatic phenotypic markers from all patients, by using a proteomic
approach.
Area di interesse identificata
Biomarkers
Biobanking
Translational research
Finanziamenti ricevuti
Titolo progetto
Disabilità cognitiva e comportamentale nelle demenze e nelle psicosi
Ente finanziatore
Fondazione CARIVERONA
Durata progetto
3 anni
Abstract del progetto
Nome
Rita Casadio, Pier Luigi Martelli
Contatti
[email protected]
[email protected]
Istituto/Dipartimento
Dipartimento di Biologia Evoluzionistica Sperimentale
Proposta di ricerca
Genetic association studies have contributed lists of genetic variations (mainly SNPs) that are
putatively related with the insurgence and/or the prognosis of the Alzheimer disease (AD). Next
generation sequencing techniques will enormously increase the amount of data on genome wide
variability associated with the AD. These data will be useful for discovering new genetic markers of
the risk and for better understanding the molecular mechanisms underlying the aetiology of the AD.
To benefit from these data, an important analysis effort has to be done in two different and
complementary directions:
1) prioritisation, for highlighting among all the reported variations those that are more likely to
be related to the AD;
2) annotation, for understanding if the variations occur in coding or in regulatory regions of the
genome and for discovering their likely molecular effects.
Computational methods that are able to integrate information from different sources in a Systems
Biology perspective are then essential. In particular we propose to implement a pipeline able to:
[DA 1]
search if the variations occur in a protein coding region, in a RNA-coding region or in a
regulatory element;
[DA 2]
predict the effect of protein mutations on localization, structure, stability, and
function;
[DA 3]
predict the correlation between protein mutations and diseases;
[DA 4]
predict the effect of protein mutations on the protein-protein and protein-DNA
interactions;
[DA 5]
identify the mRNAs targeted by miRNAs and to predict the effect of variations in
miRNAs;
[DA 6]
predict the effect of variations in regulatory regions on the binding with transcription
factors;
[DA 7]
analyse the effect of cariations on regulatory and interaction networks and on
metabolic and signalling pathways;
[DA 8]
collect, compare and store the results of previous analyses for different variations.
The Biocomputing Group of the University of Bologna developed several methods that can be
integrated in the pipeline (see www.biocomp.unibo.it). Among the others, efficient tools of interest in
this particular case are devoted to:
1) the structural and functional annotation of protein sequences (BAR: Bartoli et al. 2009);
2) the prediction of membrane protein topology (ENSEMBLE: Martelli et al. 2003);
3) the prediction of perturbations in protein stability upon mutations (I-Mutant: Capriotti et al.
2008);
4) the prediction of the correlation between protein mutations and diseases (SNPs&GO:
Calabrese et al. 2009);
5) the protein localization (BaCelLo: Pierleoni et al. 2009)
6) the prediction of protein interaction sites (ISpred: Fariselli et al. 2002) and their analysis in the
context of genome-wide interaction networks (Bartoli et al., in press)
7) the prediction of silencing effect of miRNAs (Montanucci et al. 2008).
References
Bartoli L, Montanucci L, Fronza R, Martelli PL, Fariselli P, Carota L, Donvito G, Maggi G,
Casadio R -The Bologna Annotation Resource: a non-hierarchical method for the functional
and structural annotation of protein sequences relying on a comparative large-scale genome
analysis- J Proteome Res 8:4362-4371 (2009)
Bartoli L, Martelli PL, Rossi I, Fariselli P, Casadio R -The prediction of protein-protein
interaction sites in genome-wide protein interaction networks: the test case of the human cell
cycle- Curr Prot Pept Sci (in press)
Calabrese R, Capriotti E, Fariselli P, Martelli PL, Casadio R -Functional annotations improve the
predictive score of human disease-related mutations in proteins- Hum Mutat 30:1237-1244
(2009)
Capriotti E, Fariselli P, Rossi I, Casadio R -A three-state prediction of single point mutations on
protein stability changes- BMC Bioinformatics 9 Suppl 2:S6 (2008)
Fariselli P, Pazos F, Valencia A, Casadio R -Prediction of protein-protein interaction sites in
heterocomplexes with neural networks- Eur J Biochem 269:1356-1361 (2002)
Martelli PL, Fariselli P, Casadio R -An ENSEMBLE machine learning approach for the prediction
of all-alpha membrane proteins- Bioinformatics 19:I205-I211 (2003)
Montanucci L, Fariselli P, Martelli PL, Rossi I, Casadio R -In Silico Evidence of the Relationship
Between miRNAs and siRNAs- Open Appl Inf Journal 2:9-13 (2008)
Pierleoni A, Martelli PL, Fariselli P, Casadio R -BaCelLo: a balanced subcellular localization
predictor- Bioinformatics 22:e408-e416 (2006)
Area di interesse identificata
Genome wide association studies
Nome
Contatti
Istituto/Dipartimento
Elio Scarpini, Daniela Galimberti
0255033847
[email protected]
Università di Milano, Dipartimento di Scienze Neurologiche
Proposta di ricerca
Given the rationale that AD is a multifactorial disease and that inflammation plays a role in its
pathogenesis, this proposal aims to study in detail genes encoding for inflammatory molecules, including
progranulin and chemokines, all of which located in chromosome 17q.21, both from a genetic and
functional point of view. This research will take advantage from the availability of a biobank including 500
patients with AD diagnosed according to current criteria, and a similar number of age-matched controls.
Genomic DNA have been collected for each subject. In addition, in a group of about 200 patients and
controls, serum, plasma, CSF and RNA from PBMC and CSF cells have been collected as well.
In particular, the specific objectives of this research are aimed to:
1. Carry out a thorough association analysis of the region of chromosome 17 containing PGRN and the
selected chemokine cluster (CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL13, CCL14, CCL15,
CCL16, CCL18, CCL23, CCR7, CCR10). Optimal tagging SNPs covering all the linkage blocks will be
chosen. To this aim, we will perform a power calculation using the Genetic Power Calculator
program (http://pngu.mgh.harvard.edu/~purcell/gpc/) based on sample size, the average observed
MAF, and assuming a multiplicative model and an estimated prevalence of AD in Italy of 1 case per
1000 inhabitants after 65 years of age.
2. Evaluate expression levels of the above mentioned genes in cells isolated from CSF and PBMC by
Real-Time PCR. Prioritized hits selected at aim 1) will be correlated with expression levels to test
whether these SNPs influence the level of transcription.
3. Analyze the pattern of miRNA which act as silencer of mRNA studied in aim 2). miRNA possibly
influencing the translation of the above mentioned genes will be selected by submitting queries
at the website http://microrna.sanger.ac.uk/, which contains sequences of all published mature
miRNA, together with their predicted source hairpin precursors.
4. Evalutate, by ELISA, CSF and serum levels of PGRN and chemokines.
5. Combine genetic, expression and functional data with the clinical phenotype of patients, including
longitudinal full neuropsychological testing, treatment efficacy and brain/hippocampal atrophy at
MRI, in order to identify a genetic pattern and/or novel pathogenetic mechanisms suitable to
predict which patients will likely decline more rapidly or will respond to therapy.
Centers collaborating to this project: University of Brescia (A. Padovani, D. Finazzi), IRCCS
Fatebenefratelli Brescia (G. Binetti, R. Ghidoni), IRCCS Castellanza (M. Franceschi), IRCCS S. Raffaele,
Milan (S. Cappa), University of Turin (I. Rainero). From the University of Milan: C. Mariani, R. Dominici,
Area di interesse identificata
Genetic susceptibility to Alzheimer’s disease and genome wide
association studies
Finanziamenti ricevuti
Titolo progetto
- Genetic Risk factors and peripheral biological markers of conversion
from Mild Cognitive Impairment to Alzheimer's Disease (to DG)
- Introducing new bio-markers in clinical practice for the early diagnosis of
Alzheimer disease: methodological, clinical and organisational aspects for
Ente finanziatore
Durata progetto
Abstract del progetto
the National Health System (to ES)
Progetti ( 1 and 4) parte di Programma Strategico (PS39), Italian Ministry
of Health
01-01-2009/31-12-2010
This program has two primary aims:
A. The development and validation of a multi-factorial protocol that
integrates molecular, imaging, neurophysiological, neu- ropsychological
and behavioral data for early diagnosis of Alzheimer's disease (AD), in
particular during the preclinical pha se and the prodromal stage of "mild
cognitive impairment" (MCI) when the symptoms are not severe enough
to fulfill the current diagnostic criteria for AD. The achievement of this
objective is one of the priorities of the international research com munity,
in the perspective of specific, disease-modifying therapies that are under
development.
B. Validation of this new diagnostic protocol within a regional public
health network, and evaluation of the health care, orga nization and
economic implications of its transfer to the National Health Service (NHS).
The program will be articulated into four complementary and strictly
integrated projects with the following specific aims:
PROJECT 1. Validation of genetic and biochemical markers for early
diagnosis of AD and the prediction of conversion of MCI into AD, and
design of a multivariate molecular protocol having high diagnostic and
prognostic accuracy.
PROJECT 2. Development of standard operating procedures (SOPs) and
quality control (QC) for neuroimaging (MRI, 18F-FDG PET and 18F-DDNP
PET) and clinical neurophysiological analysis (including
electroencephalography, magnetoencephalography and transcranial
magnetic stimulation) for the diagnosis of prodromic or incipient AD.
PROJECT 3. Definition of cognitive and behavioral indicators that
differentiate various forms of MCI and enable the diagno-sis of incipient
AD.
PROJECT 4. Validation of a comprehensive diagnostic protocol based on
the results obtained in the above studies, in a series of Alzheimer
Evaluation Units of Lombardia Region, and estimation of costs and
benefits of its transfer to the NHS.
Nome
Prof. Paolo Maria Rossini
Contatti
Email: [email protected]
Istituto/Dipartimento
Neurology, University “Campus Biomedico”, Rome, Italy
Proposta di ricerca
Individual evaluation of cognitive decline: definition of a low-cost non-invasive examination battery and
implementation of a predictive algorithm to identify Mild cognitive impairment.
Area di interesse identificata
Mild cognitive impairment (MCI) is a state of the elderly brain
affecting a large number of individuals mainly characterized by memory
impairment not yet encompassing the definition of dementia and, in the
majority of cases, represents a precursor of Alzheimers’ Disease (AD).
Because of MCI’s prodromal qualities do not greatly impair daily
functioning, MCI can be identified only by specific clinical and
neuropsychological evaluation. Within such a theoretical frame a reliable
marker of MCI-to-AD progression is not yet available, even if it would be
highly desirable both for research and health system purposes. Ideally,
these markers should be non-invasive, widely available and of low-cost in
order to screen out large population samples. Along this line a
combination of neurophysiological (electroencephalography, transcranial
magnetic stimulation), blood chemistries (including metals, pro- and
antioxidant compounds) and genetics are a potential candidate. . In this
way we have collected numerous studies (1-10). The line of research we
are interested in focuses on the study of healthy subjects and patients
(suffering from MCI, AD and vascular dementia – VaD-) by means of this
multidimensional investigation and will try to identify prognostic indexes
able to predict the risk of conversion from MCI to AD.
References: 1.Babiloni C, Frisoni GB, Vecchio F, Pievani M, Geroldi
C, De Carli C, Ferri R, Vernieri F, Lizio R, Rossini PM Global functional
coupling of resting EEG rhythms is related to white-matter lesions along
the cholinergic tracts in subjects with amnesic mild cognitive impairment.
J Alzheimers Dis. 2010 Jan;19(3):859-71.
2.Moretti DV, Frisoni GB, Fracassi C, Pievani M, Geroldi C, Binetti
G, Rossini PM, Zanetti O. MCI patients' EEGs show group differences
between those who progress and those who do not progress to
AD.Neurobiol Aging. 2009 Dec 16. [Epub ahead of print]
3. Babiloni C, Frisoni GB, Vecchio F, Pievani M, Geroldi C, De Carli
C, Ferri R, Vernieri F, Lizio R, Rossini PM. Global Functional Coupling of
Resting EEG Rhythms is Related to White-Matter Lesions Along the
Cholinergic Tracts in Subjects with Amnesic Mild Cognitive Impairment. J
Alzheimers Dis. 2009 Nov 17. [Epub ahead of print]
4. Moretti DV, Pievani M, Geroldi C, Binetti G, Zanetti O, Cotelli
M, Rossini PM, Frisoni GB.
Increasing hippocampal atrophy and cerebrovascular damage is
differently associated with functional cortical coupling in MCI patients.
Alzheimer Dis Assoc Disord. 2009 Oct-Dec;23(4):323-32.
5. Moretti DV, Pievani M, Geroldi C, Binetti G, Zanetti O, Rossini
PM, Frisoni GB.
EEG markers discriminate among different subgroup of patients with mild
cognitive impairment. Am J Alzheimers Dis Other Demen. 2010
Feb;25(1):58-73. Epub 2009 Feb 9.
6. Moretti DV, Fracassi C, Pievani M, Geroldi C, Binetti G, Zanetti
O, Sosta K, Rossini PM, Frisoni GB. Increase of theta/gamma ratio is
associated with memory impairment. Clin Neurophysiol. 2009
Feb;120(2):295-303. Epub 2009 Jan 1.
7. Babiloni C, Pievani M, Vecchio F, Geroldi C, Eusebi F, Fracassi C,
Fletcher E, De Carli C, Boccardi M, Rossini PM, Frisoni GB. White-matter
lesions along the cholinergic tracts are related to cortical sources of EEG
rhythms in amnesic mild cognitive impairment. Hum Brain Mapp. 2009
May;30(5):1431-43.
8. Babiloni C, Visser PJ, Frisoni G, De Deyn PP, Bresciani L, Jelic V,
Nagels G, Rodriguez G, Rossini PM, Vecchio F, Colombo D, Verhey F,
Wahlund LO, Nobili F. Cortical sources of resting EEG rhythms in mild
cognitive impairment and subjective memory complaint. Neurobiol Aging.
2008 Nov 20. [Epub ahead of print]
9 Moretti DV, Pievani M, Fracassi C, Geroldi C, Calabria M, De
Carli CS, Rossini PM, Frisoni GB. Brain vascular damage of cholinergic
pathways and EEG markers in mild cognitive impairment. J Alzheimers
Dis. 2008 Nov;15(3):357-72.
10. Rossini PM, Buscema M, Capriotti M, Grossi E, Rodriguez G, Del Percio
C, Babiloni C. Is it possible to automatically distinguish resting EEG data of
normal elderly vs. mild cognitive impairment subjects with high degree of
accuracy? Clin Neurophysiol. 2008 Jul;119(7):1534-45. Epub 2008 May 15.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Is it possible to automatically distinguish resting EEG data of normal
elderly vs. mild cognitive impairment subjects with high degree of
accuracy?
AFaR, Associazione Fatebenefratelli per Ricerca Biomedica e Sanitaria
Durata progetto
2005-2006; 2006-2007; 2007-2008; 2008-2009; 2009-2010
Abstract del progetto
To explore if Implicit function as squashing time (IFAST) based
electroencephalographic (EEG) can reliably distinguish of mild cognitive
impairment (MCI) and Alzheimer's disease (AD) subjects with
Nome
ORAZIO ZANETTI
Contatti
TEL 030 3501 358
Email: [email protected]
U.O.Alzheimer-Centro per la Memoria
IRCCS Centro S.Giovanni di Dio-Fatebenefratelli, Brescia
Istituto/Dipartimento
Proposta di ricerca
Nell’agosto 2007 un gruppo di studiosi ha proposto nuovi criteri diagnostici per la diagnosi di malattia di
Alzheimer (AD) prodromica o preclinica (Dubois et al.:Research criteria for the diagnosis of Alzheimer's
disease: revising the NINCDS-ADRDA criteria. Lancet Neurol. 2007 Aug;6(8):734-46.) incorporando, oltre ai
dati anamnestici, clinici e neurospicologici l’utilità di marker neuroradiologici (atrofia ippocampale o
ipoperfusione temporo-parietale alla PET), e liquorali (riduzione della proteina ABeta 42 ed aumento di
TAU). Dal luglio 2006 è attivo presso l’IRCCS S.Giovanni di Dio Fatebenefratelli un Ambulatorio
Traslazionale per la Memoria (ATM) che coinvolge tutte le Unità Operative cliniche e di ricerca dell’Istituto
al fine di offrire, soprattutto ai pazienti lievi un percorso diagnostico in grado di rispondere
all’interrogativo ciclopico, con risvolti laceranti, che ci veniva posto: “vorrei sapere se ho o non ho la
malattia di Alzheimer”. Nell’arco di 12 mesi sono stati valutati 144 pazienti consecutivamente afferiti all’
ATM, divisi in quattro gruppi sulla base della diagnosi clinica (utilizzando i noti criteri diagnostici NINCDSADRDA per la AD): 12 pazienti sono affetti da disturbo soggettivo di memoria (SMC, MMSE 28.0±1.1); 37
da “mild cognitive impairment” (MCI, MMSE 25.1±3.6), 55 da malattia di Alzheimer (MMSE 21.1±3.5), e 40
da demenze non Alzheimer (MMSE 21.6±5.5)(Vedi tabella). A tutti i pazienti sono state proposte le
seguenti indagini: 1) la volumetria ippocampale alla MR; 2) l’analisi visiva della PET con
fluorodesossiglucosio; 3) la concentrazione di Tau totale e di Abeta1-42 sul liquor cerebrospinale.
La sensibilità di ogni marker è più elevata (43 to 71%) per la malattia di Alzheimer rispetto all’ MCI (18 to
31%). La specificità dei marker rispetto a SMC e demenze non Alzheimer è risultata buona, essendo
superiore al’ 83% ed al 69% rispettivamente. La positività di almeno un marker presenta un’elevata
sensibilità per la AD ed un’elevata specificità per il SMC. L’utilizzo di un numero maggiore di marker (due o
tre) contemporaneamente aumenta notevolmente la specificità diagnostica a scapito però della sensibilità.
In conclusione i marker recentemente proposti per la diagnosi precoce di malattia di Alzheimer possono
essere di utilità nella pratica clinica. La valutazione longitudinale consentirà di valutare meglio il loro
“valore aggiunto” rispetto alla tradizionale diagnosi che si fonda prevalentemente sull’indagine clinica
corroborata da una semplice TAC encefalica. Per un oculato impiego delle risorse e delle nuove tecnologie
è infatti importante stabilire con precisione quali siano i pazienti (verosimilmente coloro con un disturbo
cognitivo molto lieve) che maggiormente possono trarre vantaggi da una procedura diagnostica complessa
e costosa.
Area di interesse identificata
Impiego dei makers biologici e neuro radiologici nella diagnosi
differenziale precoce
Finanziamenti ricevuti – € 20.000
(Programma strategico 2006)
Titolo progetto
Ente finanziatore
Strumenti e procedure diagnostiche per le demenze utilizzabili nella
clinica ai fini della diagnosi differenziale precoce
Ministero della Salute – Programma strategico 2006-
Durata progetto
triennale
Abstract del progetto
Nome
Sandro Sorbi
Contatti
[email protected]
tel +390554298465
Istituto/Dipartimento
Dept. of Neurological and Psychiatric Sciences, University of Florence,
Italy
Proposta di ricerca
Area di interesse identificata
Research in the field of aging and dementia is focusing on the
characterization of the early stages of cognitive decline. It has been
suggested that many patients with Mild cognitive impairment (MCI)
represents already not recognized AD patients. The purpose of the
research is to identify tools and diagnostic procedures that can identify
early stages of the disease. We will study the various domains in order to:
a) Diagnose and characterize demented patients for a proper
classification of the forms of dementia (Alzheimer's disease, fronto
temporal dementia, FTD, dementia with motor signs such as Lewy body
disease and others) b) Diagnose and characterize patients with Mild
Cognitive Impairement c) To study the neuropsychological aspects to
identified early MCI patients, both multi domain and amnesic MCI. d)
Analyze the genetic patient profile to identify the genetic risk factors
involved in the development and progression of AD.
The study of genetic factors for AD will be performed in patients with
familial forms of dementia and in patients with MCI. In addition
neuropsychological and neuroimaging evaluations will be performed for
each patients. The study will be divided into the following phases: analysis of autosomal dominant genetic mutations: new families affected
by AD will be identified and characterized clinically and genetically. The
goal will be to identify new causative mutations. the Initial screening of
molecular diagnosis will be a detailed genetic screening of the genes
associated with familial form of the disease, presenilins (PS-1 and PS-2)
and the amyloid precursor protein (APP) - Search for other possible
factors of genetic susceptibility will be performed on both familial and
sporadic AD patients. It will be studied allelic variations of genes
potentially involved in chromosomal regions of interest. It will also be
carried out the study of Apolipoprotein E genotype (ApoE), considered
today the only susceptibility factor associated with the disease.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
“Strumenti e procedure diagnostiche per le demenze utilizzabili nella
clinica ai fini della diagnosi precoce e differenziale, della individuazione
delle forme a rapida o lenta progressione e delle forme con risposta
ottimale alle attuali terapie”
MINISTERO DELLA SALUTE , RICERCA FINALIZZATA 2006
Durata progetto
6/1/2008-6/4/2010
Abstract del progetto
The main aim of the project is to integrate the results of neurocognitive,
neuropsychological, neuroimaging and genetic studies as markers for the
differential diagnosis of early dementia.
Titolo progetto
GENETIC STUDY ON ALZHEIMER'S DISEASE AND THE INVOLVEMENT OF
OTHER NON-GENETIC FACTORS THAT MAY INFLUENCE THE CLINICAL
EXPRESSION OF THE DISEASE
Ente finanziatore
Progetto di ricerca scientifica d'Ateneo (ex quota 60%)
Durata progetto
2009-2010
Abstract del progetto
GENETIC STUDY ON ALZHEIMER'S DISEASE
Titolo progetto
"Analisi dei fattori genetici coinvolti nelle malattie neurodegenerative:
Utilizzo e applicazione di tecnologie di avanguardia per lo studio della
malattia di Alzheimer, della demenza frontotemporale e della malattia di
Parkinson" /*
Ente finanziatore
COMPAGNIA SAN PAOLO
Durata progetto
2008-2010
Abstract del progetto
GENETIC STUDY ON ALZHEIMER'S DISEASE
Proposta di ricerca
Area di interesse identificata
Aim of the study is to determine the clinical and genetic features of
movement disorders in patient affected by dementia and the clinical
features of cognitive decline in patients affected by movement disorders.
All the patients recruited will be assessed at baseline and during the
follow-up with selected clinical scales and neuropsychological tests.
Beyond movement disorder and cognitive deficit, attention will be paid to
behavioural disturbances, mood disorders and impulse control disorders
in these patients.
Patients with a familial history of movement disorders or dementia will
undergo genetic analysis on the genes implicated in autosomal dominant
forms of Parkinson disease including PARK1, PARK4, SNCA and LRRK2.
All the patients recruited will be submitted to FP-CIT-SPECT in order to
verify and to measure the presence of nigrostriatal pathway
C.
Nome
Flavio Nobili, Guido Rodriguez
degeneration.
We plan to enrol 50-100 patients affected by those neurodegenerative
disorders that may be characterized by movement disorders and
cognitive decline: Parkinson’s disease, Alzheimer’s disease with
parkinsonism, Frontotemporal dementia, Lewy body disease, vascular
dementia, corticobasal degeneration and progressive supranuclear palsy.
Finanziamenti ricevuti
Titolo progetto
“ Epidemiologia e sviluppo di strumenti di integrazione socio-sanitaria
nella gestione dei pazienti affetti da Malattia di Parkinson e da altre
malattie degenerative con concomitante disturbo motorio e declino
cognitivo”
Ente finanziatore
MINISTERO DELLA SALUTE, RICERCA FINALIZZATA 2007
Durata progetto
12/2009-12/2010
Abstract del progetto
Main aim of the project is to analyze and develop in detail the natural
history of Parkinson's disease and Parkinsonism due to cognitive decline
and behavioural disturbances. Epidemiological study is of fundamental
importance in developing diagnostic and therapeutic strategies and new
methods of integrating social and health care.
In particular we will identify and recruit individuals with Parkinson's
disease and classical subjects with parkinsonism linked to cognitive
decline, it will estimate the incidence of dementia in the two types of
disturbances. Will be performed genetic analysis of genes related to
Parkinson's disease in people with familiarity with the disease. All
patients will undergo a battery of standardized tests (Unified Parkinson's
Disease Rating Scale and Hoehn and Yahr and neuropsychological tests)
and FP-CIT-SPECT imaging.
Contatti
Istituto/Dipartimento
[email protected]; [email protected]
Tel: 010 3537568; Fax: 010 5556893; cell: 333 8331613
Neurofisiologia Clinica, Dip. di Neuroscienze, Oftalmologia e Genetica,
Università di Genova
Proposta di ricerca
A.
‘A multimodality index for the early diagnosis of Alzheimer’s disease’
Area di interesse identificata
Early diagnosis
Finanziamenti ricevuti
Titolo progetto
‘Descripa’ (http://www.descripa.eu/)
Ente finanziatore
EU FP-5
Durata progetto
4 years
Abstract del progetto
Titolo progetto
The DESCRIPA study aims to develop screening guidelines and clinical
criteria for Alzheimer’s disease in non-demented subjects. The clinical
criteria will be based on a prospective cohort study of non-demented
subjects from a memory clinic. The screening guidelines will be based on
a meta-analysis of prospective population-based cohort studies in
Europe.
‘ICTUS’ (Neuroepidemiology. 2007;29(1-2):29-38.)
Ente finanziatore
EU FP-5
Durata progetto
3 years
Abstract del progetto
Observational study of cognitive and behavioural effects of
acetylcholinesterase inhibitors in patients with mild to moderate
Alzheimer’s disease
Finanziamenti ricevuti
Titolo progetto
‘Descripa’ (http://www.descripa.eu/)
Ente finanziatore
EU FP-5
Durata progetto
4 years
Abstract del progetto
The DESCRIPA study aims to develop screening guidelines and clinical
criteria for Alzheimer’s disease in non-demented subjects. The clinical
criteria will be based on a prospective cohort study of non-demented
subjects from a memory clinic. The screening guidelines will be based on
a meta-analysis of prospective population-based cohort studies in
Europe.
‘ICTUS’ (Neuroepidemiology. 2007;29(1-2):29-38.)
Titolo progetto
Ente finanziatore
EU FP-5
Durata progetto
3 years
Abstract del progetto
Observational study of cognitive and behavioural effects of
acetylcholinesterase inhibitors in patients with mild to moderate
Alzheimer’s disease
BACKGROUND. During the last decade, there has been a great effort to improve diagnostic accuracy of
Alzheimer’s disease (AD) since the earliest stages. The main advances have been observed with
morphological Magnetic Resonance (MR) and functional (FDG-PET) brain imaging, and with Cereborspinal
fluid (CSF) assays of Tau protein and β amyloid fragments. The accuracy of these tools is rather good when
comparing patients with overt AD dementia versus healthy controls, but decreases in the very early stages
of the disease. In these stages, the subjects still maintain full autonomy in everyday life, but subtle deficit in
memory and/or other cognitive domains can already be shown by cognitive tests (Mild Cognitive
Impairment, MCI). Moreover, some patients complain of memory disturbances that cannot be confirmed
by cognitive assessments (subjective memory complaints, SMC) but that may sometimes be prodromic of
AD.
Pharmacological research is currently very active in trying to identify drugs that can modify the
natural course of the disease (‘disease-modifying’). While they are tested mainly in overt AD dementia at
present, it is obvious that effective drugs will be used as soon as possible in the course of the disease, i.e.,
at the MCI stage, or even earlier.
Thus, an early diagnosis is of paramount importance to start effective treatment as soon as these
drugs will be available. Yet, individual diagnostic accuracy of functional and morphological neuroimaging
and CSF assays in these early stages is not fully satisfactory versus healthy controls but even more versus
the other commonest forms of dementia, such as dementia with Lewy bodies (DLB), frontotemporal
dementia (FTD) and vascular dementia (VaD). The more recent amyloid PET imaging has added value to
neuroimaging, showing very high sensitivity but limited specificity versus healthy controls, DLB and
Parkinson’s disease dementia.
The reasons of incomplete accuracy are complex and partially understood. Genetic heterogeneity,
environmental factors, and phenotipic variants of AD may explain incomplete sensitivity on the one hand.
On the other hand, overlapping atrophy, hypometabolic and biochemical patterns with FTD, DLB and VaD
may account for incomplete specificity of MR, PET, and CSF assays, respectively.
The idea that the ‘shadow’ area of each diagnostic tool could be 'unmasked' by the other tools has
moved researchers to join together the data deriving from more than one modality. Some work has already
been done, showing an increase accuracy when two examinations are employed together. In this context,
the ‘core’ cognitive deficit of AD, i.e., verbal episodic memory delayed recall, has also been included.
Previous attempts to obtain combination indexes include neuropsychology and PET (1-2), neuropsychology,
PET and Apolipoprotein E (ApoE) genotype (3), neuropsychology and MR imaging (4), Single Photon
Emission Computed Tomography (SPECT) and MR imaging (5), CSF markers and SPECT (6). However, the
data are still largely incomplete, especially regarding the very early stages of the disease.
AIM. To test the hypothesis that joining together the information deriving from MR and FDG-PET imaging,
memory tests, CSF assays and genetic predisposing factors (such as the ApoE genotype) can improve
diagnostic accuracy in patients with AD dementia at the MCI stage. A single index could be achieved with
integrated accuracy deriving from each tool.
METHODS. Patients. A multicentric approach is needed to reach large patient series in a limited time.
Patients with MCI of the amnestic type (aMCI), as well as age-matched healthy controls, should be
recruited by Centres with proven expertise in dementia diagnosis and management. These subjects should
undergo full neuropsychological examination (common minimum battery to be agreed), MR imaging, and
FDG-PET. CSF assays are performed in MCI patients and in that part of healthy controls giving the informed
consent for this invasive procedure, providing Ethic Committees approve this manoeuvre in healthy
subjects. Administration of radiopharmaceuticals to healthy subjects must also be allowed by Ethic
Committees and by the National Authority governing radiation exposure to healthy subjects for research
purposes.
Then, both patients and controls are regularly followed by means of clinical and neuropsychological
examinations on yearly basis for at least three years to pick up those MCI patients matching the current
criteria for the diagnosis of AD or other dementias. Other study groups are represented by patients
affected with FTD, VaD or LBD in a mild stage (i.e., MMSE score >20), recruited by the same centres,
following the internationally accepted guidelines.
Image analysis. Several software are available to automatically segment the whole brain or specific brain
regions, such as the hippocampus and the inferior parietal lobule, that better distinguish AD patients and
controls. These software can either run both MR and PET images (such as Statistical parametric Mapping,
SPM) or specifically one of the two (such as the boundary shift integral for MR and ‘Neurostat’ for PET). As
for MR images, the output could be a volumetric index. The same applies to FDG-PET where a metabolic
index can be automatically computed. For both MR and PET, other indexes expressing the probability to
have maximum or minimum atrophy/hypometabolism versus a control group can be computed.
Statistical analysis. Statistics should rely on a multidimensional approach able to identify the best
combination of indexes that distinguish between AD patients (at MCI stage) and either healthy controls or
DLB, FTD, and VaD, respectively. Stepwise and discriminant approaches are favourites.
EXPECTED RESULTS. An index able to discriminate AD from healthy controls and from the other
commonest foms of dementia with very high accuracy, i.e., >95%. This index should work, even if with a
more limited accuracy, in those cases where just some of the requested information is available, for
instance just with neuropsychology, ApoE, MR and PET images, without CSF biomarkers. Once established,
such an index may be applied also to subjects with SMC to unveil AD pathology, as well as to screen
population at high risk of AD, such as direct relatives of AD patients.
REFERENCES. (1) Anchisi D, et al. Heterogeneity of brain glucose metabolism in mild cognitive impairment
and clinical progression to Alzheimer disease. Arch Neurol 2005;62:1728-33. (2) Nobili F, et al. Principal
component analysis of FDG PET in amnestic MCI. Eur J Nucl Med Mol Imaging 2008;35:2191-2202. (3)
Mosconi L, et al. MCI conversion to dementia and the APOE genotype. A prediction study with FDG-PET.
Neurology 2004;63:2332-40. (4) Visser PJ, et al. Medial temporal lobe atrophy and memory dysfunction as
predictors for dementia in subjects with mild cognitive impairment. J Neurol 1999;246:477-85. (5) El Fakhri
G, et al. MRI-guided SPECT perfusion measures and volumetric MRI in prodromal Alzheimer disease. Arch
Neurol. 2003;60:1066-72. (6) Okamura N et al. Combined Analysis of CSF Tau Levels and
[(123)I]Iodoamphetamine SPECT in Mild Cognitive Impairment: Implications for a Novel Predictor of
Alzheimer's Disease. Am J Psychiatry. 2002;159:474-6. (7) Barnes J, et al. Automatic calculation of
hippocampal atrophy rates using a hippocampal template and the boundary shift integral. Neurobiol Aging.
2007;28:1657-63. (8) Ishii K, et al. Statistical brain mapping of 18F-FDG PET in Alzheimer's disease:
validation of anatomic standardization for atrophied brains. J Nucl Med. 2001;42:548-57.
Nome
Professor Paolo Caffarra, M.D.
Contatti
e-mail: [email protected]
phone/fax: 0039-0521-704116
Department of Neuroscience
University of Parma
Via Gramsci 14
43100 Parma
Italy
Istituto/Dipartimento
Proposta di ricerca
Area di interesse identificata
The need of an early diagnosis is essential for promoting a strategic
pharmacological and non-pharmacological intervention on Alzheimer’s
disease (AD). The identification of cognitive and neurobiological
markers which might represent salient pre-clinical predictors of the
insurgence of AD is essential nowadays. Our research focuses on the
discovery of cognitive instruments useful for a differential diagnosis of
AD, such as memory and executive tasks, and neuroimaging markers
using different techniques, such as magnetic resonance imaging (MRI)
and positron emission tomography (PET). Recently, we have set up an
innovative and computerized experimental procedure to score semantic
abilities in patients suffering from Mild Cognitive Impairment (MCI) and
mild AD (Arango-Lasprilla et al., 2007; Ahmed et al., 2008) , investigating
also the functional and metabolic neural substrates using MRI and F18FDG-PET. Different aspects of structural MRI will be observed, such as
volumetric analysis, diffusion tensor imaging (DTI), imaging of default
mode at rest and imaging of the deposition of iron in memory-related
critical brain regions, i.e. hippocampus. In particular, the observation of
iron deposition in the brain of patients at risk for AD, represents an
innovative approach in the neuroimaging field of dementia (Ding et al.,
2009). F18-FDG-PET constitutes an effective technique in the monitoring
of early Alzheimer’s disease progression (Fouquet et al., 2009).
Cognitive and neuroimaging follow-up data will be informative on the
identification of specific possible markers of conversion (Devanand et
al., 2007; McGeown et al., 2009). The role of the polymorphism of APOE
is observed and correlated with clinical progression of the disease,
cognitive performance and neuroimaging pattern.
References:
Ahmed S, Arnold R, Thompson SA, Graham KS, Hodges JR. (2008).
Naming of objects, faces and buildings in mild cognitive impairment.
Cortex, 44, 746-52.
Arango-Lasprilla JC, Cuetos F, Valencia C, Uribe C, Lopera F. (2007).
Cognitive changes in the preclinical phase of familial Alzheimer's
disease. J Clin Exp Neuropsychol. 29, 892-900.
Devanand DP, Pradhaban G, Liu X, Khandji A, De Santi S, Segal S,
Rusinek H, Pelton GH, Honig LS, Mayeux R, Stern Y, Tabert MH, de Leon
MJ. (2007). Hippocampal and entorhinal atrophy in mild cognitive
impairment: prediction of Alzheimer disease. Neurology, 68, 828-36.
Ding B, Chen KM, Ling HW, Sun F, Li X, Wan T, Chai WM, Zhang H, Zhan
Y, Guan YJ. (2009). Correlation of iron in the hippocampus with MMSE in
patients with Alzheimer's disease. J Magn Reson Imaging, 29, 793-8.
McGeown, W.J., Shanks, M.F., Forbes-McKay, K.E., Venneri, A. (2009).
Patterns of brain activity during a semantic task differentiate normal
aging from early Alzheimer's disease. Psychiatry Res., 173, 218-27.
Finanziamenti ricevuti
Titolo progetto
“CORRELATI FUNZIONALI E STRUTTURALI DELLE FUNZIONI COGNITIVE
NELLA FASE PRE-CLINICA E CONCLAMATA DELLE DEMENZE
DEGENERATIVE”
Ente finanziatore
Cassa di Risparmio di Parma e Piacenza
Durata progetto
24 mesi
Abstract del progetto
The present project aims to investigate the neurobiological substrates
associated with the insurgence and progression of Alzheimer’s disease
(AD). In particular, the attention will be focused on the study of brain
regions more vulnerable and affected by AD pathology and responsible
for AD symptoms, such as hippocampal, posterior cingulate and parietal
circuits involved in memory function. The project will concentrate on the
identification of distinct neurofunctional and structural pattern which
characterize the different forms of dementia, in order to establish the
effectiveness of neuroimaging techniques in the diagnostic process of
dementia. Particular attention will be give on the study of neuroimaging
correlates of Mild Cognitive Impairment as this is considered a critical
phase in the progression to dementia: the identification of typical preclinical neuroimaging markers of AD will provide the possibility to act an
early and effective intervention.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Stefano F. Cappa
DIBIT via Olgettina 58
20132 Milano tel 0226434887
[email protected]
Vita Salute University and
San Raffaele Scientific Institute, Division of Neuroscience
Proposta di ricerca.
Our multi-disciplinary reaserach unit, combines expertises from different backgrounds (neurology,
neuropsychology, linguistics, cognitive psychology, epistemology), with the general aim to investigate the
neural mechanisms of language and high-order cognition. The experimental approaches include
behavioural studies in normal subjects and in neurological patients, brain imaging using magnetic
resonance techniques and positron emission tomography, and neurophysiological investigations based on
evoked responses and transcranial magnetic stimulation. In the field of dementia we are leading a number
of investigation in the areas of linguistic function, memory and social neuroscience. Besides the theoretical
interest , these studies are of relevance for issues such as early diagnosis (in particular distinction from
healthy aging), differential diagnosis among different causes of degenerative dementia, and assessment of
treatment effect. The studies are typically based on the development of innovative behavioural testing
procedures, combined with multimodal imaging, biomarkers and genetic investigations, and are largely
based on collaboration amiong multiple clinical and research units, both within the San Raffaele
Foundation and with other national and international institution.
Area di interesse identificata
Cognitive neuroscience of dementia: clinical and theoretical aspects
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
The reseach has been supported by grants from the Ministry of Health,
the MIUR, the EEC and Human Frontiers organization
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Daniela Perani
02-26432224 -2223
[email protected]
V-S San Raffaele University, Division of Neuroscienze and Nuclear
Medicine Dpt. San Raffaele Scientific Institute
Proposta di ricerca
Area di interesse identificata
Studies are aimed at investigating with PET the brain functional
parameters and neurotransmission systems in neurological and
psychiatric diseases :
18
F-FDG is used for the evaluation of brain glucose metabolism and 11CPIB for amyloid deposition. Present protocols include dementias (AD and
FTLD, LBD, PDD) in early and pre-clinical phase, including in Mild Cognitive
Impairment.
11
C-PK and its developed forms are used in the study of
neuroinflammation. Present protocols include Parkinson’s disease and
parkinsonisms such as Lewy Body Dementia, prion diseases, and
Amiotrophic Lateral Sclerosis. To this aim mathematical models for
quantification of activity are under development.
11
C-Raclopride and 11C-Fe CIT for the study of dopaminergic system.
Present protocols include Parkinson’s disease and parkinsonisms,
particularly in early phase.
Voxel-based methods for automatic
quantification are under study validation.
11
C MP4 for measuring the AchE activity, in AD and MCI subjects in order
to obtain correlations between reduction of enzymatic activity and
cognitive status. To this aim mathematical models for quantification of
activity are under development.
11
C-MDL for 5HT2 serotonin receptors and 11C-raclopride. Present
protocols include psychiatric conditions such as obsessive compulsive
disorders and depression
11
C-FMZ for the evaluation of the gabaergic system. Present protocols
include movement disorders such as dystonias (genetic and sporadic
cases)
Finanziamenti ricevuti
Titolo progetto
1. Diagnosis by Statistical and Intelligent Systems (ICT 2009, 5.3).
2. Molecular imaging for the early diagnosis and monitoring of
Alzheimer’s disease in old individuals with cognitive disturbances:
Ente finanziatore
Durata progetto
Abstract del progetto
an ADNI-compatible prospective study.
3. Identification of diagnostic biomarkers in Amyotrophic Lateral
Sclerosis patients, and development of related computational
methods.
1. Comunità Europea
2. Ministero della Sanità
3. Ministero della Sanità
1. 2010-2013
2. 2010-2012
3. 2008-2010
1. Validazione di nuovi metodi voxel-based e pattern recognition
per il neuroimaging funzionale PET, SPECT e strutturale MRI nella
diagnosi precoce e nella diagnosi differenziale delle malattie
neuroddegenerative associate a demenza.
2. Studio dei correlati neurofunzionali, anatomici e molecolari nel
disturbi cognitivi lievi.
3. Studio di biomarker in vitro e in vivo nella Sclerosi Laterale
Amiotrofica.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Maurizio Popoli
Tel: +390250318361
E-mail: [email protected]
Center of Neuropharmacology, Department of Pharmacological Sciences,
CEND, University of Milan
Proposta di ricerca
Area di interesse identificata
3C. Global proteomics of human fibroblasts for the identification of
early biomarkers for dementia and Alzheimer’s Disease
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Identification of biomarkers for Alzheimer’s dementia: genomics and
proteomics of neurodegeneration
Ministero della Salute
Durata progetto
2004-2006
Abstract del progetto
Ente finanziatore
Main object of the present project was the identification of genetic and
biological markers for Mild Cognitive Impairment (MCI) in order to
identify a strategy useful for an early diagnosis and for the individuation
of subjects at risk for the development of Alzheimer’s disease.
Neurodegeneration, physiological cerebral aging and dementia: an
integrated approach for a early diagnosis, evolution predictors, and for an
advanced care organization.
Ministero della Sanità
Durata progetto
2000-2003
Abstract del progetto
Aim of this project was the investigation of biological, cellular, genetic
and neurotoxic processes underling dementia for the identification and
definition of predictive markers of pathology in the general population
and in presence of risk factors.
Titolo progetto
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Gianfranco Spalletta
Via Ardeatina, 306 -00179 Rome
06-51501575
[email protected]
IRCCS Santa Lucia Foundation/Department of Clinical and behavioral
neurology - Neuropsychiatry laboratory
Proposta di ricerca
Area di interesse identificata
Parkinson disease and parkinsonisms
Finanziamenti ricevuti
Titolo progetto
Analysis of cognitive and neuropsychiatric disturbances in Parkinson
disease and parkinsonisms: implications for early diagnosis
Ente finanziatore
Italian Ministry of Health
Durata progetto
22/12/2008 - 22/12/2011
Abstract del progetto
The prevalence rate for neuropsychiatric symptoms of depression,
anxiety, and hallucinations in subjects with Parkinson Disease (PD) has a
very wide range. This suggests how undefined the results of studies done
up till now are. Moreover, longitudinal studies suggest that the majority
of subjects with PD could develop dementia during the course of the
disease. This study aims at evaluating the differential features of
cognitive and neuropsychiatric disturbances in subjects with PD and other
Parkinsonisms at the onset of the disease vs. advanced phases. A crosssectional model could help to understand the phenomena changing along
time in a time-limited project. If psychopathological disturbances
associated with Parkinson disease are difficult to be studied, those that
accompany other motor disturbances are almost completely unknown.
Therefore, it is urgent to fill this gap in knowledge with studies aimed at a
better diagnostic typing between PD, LBD and other degenerative and
vascular parkinsonisms. Another important issue concerns the validity of
diagnostic criteria of DSM-IV for mood disturbances and anxiety
disturbances in subjects with motor disturbances. Thus, in this crosssectional study, 300 patients affected by motor disturbances will be
recruited. In particular, we will include 150 subjects with idiopatic
Parkinson Disease (PD) of which 50 at onset and 100 in the advanced
phase, 50 subjects with Dementia with Lewy Bodies (DLB); 50 subjects
with vascular PD; 25 subjects with Progressive Supranuclear Palsy (PSP);
20 subjects with corticobasal degeneration (CBD) and 10 subjects with
multisystemic atrophy (MSA). Patients will be assessed using a clinical
battery comprising: a neurological battery including socio-demographic
and clinical history, Unified Parkinson’s Disease Rating Scale (UPDRS), and
Hoehn and Yahr scale. The neuropsychological battery will include: Mini
Mental State Examination, Rey 15-word test, reduced Wisconsin Card
Sorting, reduced Stroop Color-Word test, Verbal and visual-spatial Nback, Verbal semantic fluency, Verbal phonological fluency, Sentence
construction, Immediate and deferred copy of Rey figure. The
neuropschiatric battery will include: Psychiatric diagnosis according with
the DSMIV-TR (in this case we will use both the standard diagnostic
criteria and modified diagnostic criteria proposed by the Starkstein group
in order to improve the neuropsychiatric diagnosis in patients with motor
disturbances), Beck Depression Inventory, Toronto Alexithymia Scale,
Penn Emotional Recognition Test, Snaith-Hamilton Pleasure Scale. In
another part of the study we will include 20 idiopatic PD patients treated
with Deep Brain Stimulation (DBS). They will be clinically evaluated with
UPDRS Part III and with the neuropsychological and neuropsychiatric
battery above-mentioned. The evaluation of subjects with stimulator
implants will be carried out without any treatment and in 3 different
phases: 1) during proper and stable antiparkinsonian therapy and
Stimulator on, 2) during pharmacological therapy and DBS stimulator off
since 90 minutes, 3) absence of pharmacological therapy with DBS
stimulator on. The neuropsychological and behavioural tests will be
carried out in a random way on different days. In the subgroup of
patients with DBS the evaluation for depression will be carried out with a
visual-analogical scale to instantly capture the severity of the depression
in each of the 4 experimental conditions. Results of studies in this field
will allow to propose new diagnostic criteria for neuropsychiatric and
behavioural disturbances which will help clinicians in the early and
differential diagnosis of movement disorders allowing also a better
treatment. The research on patients in treatment with DBS will allow a
better understanding of the features of non motor symptoms and
directions towards treatments. The study will also provide a picture of the
differential expression of cognitive versus neuropsychiatric symptoms
within each pathological entity. Such achievements will be possibly imply
a reduction of the burden caused by these diseases on the patient’s
family, as well as a decrease of the social and heath costs.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Gianfranco Spalletta
Via Ardeatina, 306 - 00179 Rome
O6-51501575
[email protected]
IRCCS Santa Lucia Foundation/Department of Clinical and behavioral
neurology - Neuropsychiatry laboratory
Proposta di ricerca
Area di interesse identificata
Mild Cognitive Impairment and Dementias
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Cognitive and behavioural indicators of conversion from Mild Cognitive
Impairment to neurodegenerative dementia and development of
cognitive rehabilitation protocols
Italian Ministry of Health
Durata progetto
01/01/2009 - 31/12/2011
Abstract del progetto
According to a four-group classification, the MCI amnestic single domain
is characterized by a selective deficit of amnesic performances, multidomain amnestic MCI presents memory impairment and at least one
other cognitive domain impairment, nonamnestic single domain MCI is
associated with the compromise of one nonamnestic domain, and
nonamnestic multi-domain MCI is associated with the compromise of two
or more nonamnestic cognitive domains. The hypothetic usefulness of
subdividing MCI to identify specifically the type of dementia to which the
patient will convert has partially failed. However, there is solid evidence
that multi-domain MCI has the highest rate of conversion in dementia.
Although there is convergence on the description of the high risk of
developing AD in subjects with chronic depression, often associated with
hippocampal atrophy, there are also studies which clarify with sufficient
reliability that other neuropsychiatric disturbances may predict
conversion to AD. Regarding the fact that therapeutic strategies for the
pathogenesis of AD have failed up till now, and considering the high risk
of patients with MCI to develop AD, a revision of the clinical-diagnostic
criteria of AD has been recently proposed. This revision makes AD
diagnosis similar to those of amnesic MCI with the exception of biological
and neuroradiological markers recognized as specific for AD diagnostic
purposes. Unfortunately, the specific neuropsychiatric symptoms of AD
were not included even in this recent diagnostic revision proposal. This
research project will attempt to identify clinical characteristics useful for
predicting the risk of developing neurodegenerative dementia from the 4
different forms of MCI, the type of dementia more frequently associated
with each clinical characteristics, the progression severity of the illness; d)
the effect of caregiver support on the illness progression, and the efficacy
of cognitive rehabilitation on every subtype of clinical characteristics. The
method of open study on cognitive rehabilitation will give an early clinical
indication of the potential of this treatment. One hundred patients
diagnosed with MCI will be included in the study. The patients included
will carry out an in depth anamnestic evaluation and a
diagnostic\categorical evaluation for MCI subtype. In addition, categorical
neuropsychiatric disorders of depression, psychosis, and apathy will be
assessed. Dimensional neuropsychiatric symptom severity will be
measured with a battery of tests including: Neuropsychiatric Inventory-12
items, CERAD Disforia, and Dementia Apathy Interview and Rating. The
age at onset of each behavioural disturbance and the first memory deficit
will be accurately evaluated. Moreover, To obtain a global index of
cognitive impairment, we will administer the MMSE. The Mental
Deterioration Battery and other cognitive task will be used to assess
individual cognitive domains. A subsample of 30 MCI patients will be
treated with 12 weeks cognitive rehabilitation. Also, a subsample of
caregivers of 30 MCI patients will undergo psychometric evaluation at
baseline and at the three and six-month follow-up. All subjects will be
included during the first 6 months period of the project. All diagnostic,
cognitive and neuropsychiatric tests will be administered every six
months until 18 months from the first evaluation.
This project will allow a more correct clinical-diagnostic subclassification
of MCI subgroups in order to make the clinical subtypes of MCI more
homogeneous both on the diagnostic level and the biological correlates.
In particular, this most reliable subclassification should help to fasten the
recognition of the disorders and the differential diagnosis, thus
facilitating a more precocious and specific treatment of patients who will
develop the different types of dementia. This project will also suggest the
potential role of cognitive rehabilitation therapy and caregivers support
on the cognitive and behavioural symptom progression of MCI. To obtain
this result potentially means a reduced cost of the management of
dementia for the national health service and a better outcome of the
illness for patients.
Nome
Paolo Calabresi
Contatti
Clinica Neurologica, Università degli Studi di Perugia, Ospedale S. Maria
della Misericordia, 06132 Perugia, Italy
e-mail: [email protected]
Istituto/Dipartimento
Clinica Neurologica, Laboratori di Neurologia sperimentale,
Dipartimento di Specialitá Medico Chirurgiche e Sanitá Pubblica,
Università degli Studi di Perugia
Proposta di ricerca
Area di interesse identificata
Electrophysiological study of striatal physiology in normal conditions
and in experimental parkinsonism: use of genetic and pathogenetic
animal models.
Pre-clinical research is conducted at the Laboratory of Experimental
Neurology of the Clinica Neurologica at the Hospital S. Maria della
Misericordia in Perugia. The activities focus on the study of important
neurological pathologies such Parkinson’s disease (PD) and other
neurological and neurodegenerative conditions (Huntington disease,
Alzheimer disease, Multiple sclerosis, stroke, epilepsy) by means of in
vitro animal models.
The main topic of the laboratory is the study of the mechanisms
involved in the physiopathology of such diseases by means of up to
date electrophysiology and behavioural techniques in tight
collaboration with the Laboratories of Neurophysiology located at the
Fondazione S. Lucia in Rome.
Motor and non-motor symptoms occurrence in PD represent the
downstream effect of a pathological cascade resulting in the
degeneration of dopaminergic neurons of the substantia nigra pars
compacta (SNpc) projecting to the nucleus striatum. In this nucleus
physiological activation of neuronal pathways are able to produce longterm depression (LTD), long-term potentiation (LTP) and
“depotentiation” of synaptic transmission that are all disrupted by the
PD-related pathological process. While Levodopa (L-DOPA) remains the
gold standard of symptomatic therapy of PD it also causes severe longterm side effects (L-DOPA-induced dyskinesia, LID). The cellular and
molecular mechanisms of LID formation and maintenance have started
to be elucidated.
For the study of these phenomena we take advantage of several animal
models of PD. A well accepted model is obtained with the injection of 6hydroxydopamine (6-OHDA) into the SN causing permanent DAdenervation of the ipsilateral striatum mimicking the symptoms of the
human PD. The in vitro model of PD by acute application of the
neurotoxin rotenone that affects the complex I of mitochondrial chain
is an important tool to explore mitochondrial function linked to PD. In
order to assess specific aspects of the pathogenesis of PD, we employ
transgenic animal lines expressing different mutant forms of αsynucleins such as A53T-α-synuclein mice and mice expressing human
α-synuclein lacking the C-terminal 20 amino acids (α -Syn120).
Furthermore, in order to study and to prevent LID development
(priming) and maintenance in PD models we focussed on the
modulation of the Ras-ERK pathway by the use of lentiviral vector (LV)mediated expression of either small harpin RNAs (shRNAs) specific for
Ras-GRF1 and DARPP-32.
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Titolo progetto
Ente finanziatore
Durata progetto
Titolo progetto
Ente finanziatore
Durata progetto
Titolo progetto
Ente finanziatore
Durata progetto
Titolo progetto
Ente finanziatore
Durata progetto
Titolo progetto
Ente finanziatore
Durata progetto
Titolo progetto
Ente finanziatore
Durata progetto
Nuovi approcci terapeutici per le discinesie indotte dal trattamento con
L-DOPA in un modello sperimentale di Parkinson: ruolo delle subunità
del recettore NMDA e della via molecolare Ras-ERK
Prin 2008
Two years
LIDYAS - An innovative approach to treat levodopa-induced dyskinesia
based on targeting striatal intracellular signalling
Progetto San Paolo di Torino - Coordinatore: Istituto Nazionale di
Neuroscienze (INN)
Three years
A multidisciplinary approach to test the therapeutic potential of neural
stem cell transplantation in preclinical mouse models of Parkinson’s
disease
Progetto Strategico ex art56 Centro San Raffaele del Monte Tabor
Two years
Meccanismi di danno neuronale alla base della neurodegenerazione
nelle patologie dei gangli della base
Progetto Finalizzato 2005
Two years
SYNSCAFF - Synaptic scaffolding proteins orchestrating cortical synapse
organisation during development.
SIXTH FRAMEWORK PROGRAMME - PRIORITY [LSH-2003-2.1.3-5]
[Cortical development] Proposal/Contract no.:511995
Four years
Mechanisms of L-DOPA-induced dyskinesia in an experimental model of
Parkinson's Disease: focus on NMDA receptors
Ministero dell’Istruzione dell’Università e della Ricerca (Prin 2005)
Two years
Ruolo dell’adenosina nella trasmissione sinaptica eccitatoria striatale:
interazione con il sistema dopaminergico
Sigma Tau
One years
Titolo progetto
Ente finanziatore
Durata progetto
Titolo progetto
Ente finanziatore
Mechanisms underlying cell-type specific vulnerability of striatal
neurons: implications for huntington's disease
Telethon GGP02035
Two years
Durata progetto
REPLACES - Restorative Plasticity At Corticostriatal Excitatory Synapses
Comunità Europea - HEALTH-2007-2.2.1-7 - Grant agreement no.:
222918
Four years
Titolo progetto
Ente finanziatore
Durata progetto
Imaging and biological markers of disease progression
Progetto STRATEGICO Ministero della Sanità - Sottoprogetto 5
Two years
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Lucilla Parnetti
Clinica Neurologica, Università degli Studi di Perugia, Ospedale S. Maria della
Misericordia, 06132 Perugia, Italy
Tel: +39 075 578 3545; Fax: +39 075 578 3621.
[email protected]
Clinica Neurologica, Centro Disturbi della Memoria – Unità Valutativa
Alzheimer - Laboratorio di Neurochimica Clinica, Dipartimento di
Specialitá Medico Chirurgiche e Sanitá Pubblica, Università degli Studi di
Perugia
Proposta di ricerca
Area di interesse identificata
Cerebrospinal fluid biomarkers in Alzheimer’s disease and other
neurodegenerative diseases
Clinical research in the Laboratory of Neurochemistry is performed in
close contact with the Center for Memory disturbancies, the Center for
Movement disorders and the Center for Demyelinating disease of the
Hospital S. Maria della Misericordia in Perugia. In the Laboratory of
Neurochemistry, a biobank of cerebrospinal fluid (CSF) and plasma
samples from patients with neurological diseases is available, making
feasible retrospective studies on several pathologies such as Alzheimer’s
disease (AD), Parkinson’s disease (PD) and other neurodegenerative
diseases.
The main topic of the laboratory is the study of CSF biomarkers for the
early and/or differential diagnosis of AD. The classical biomarkers Aβ1-42,
tau protein (t-tau), and phosphorylated tau 181 (p-tau) are routinely
assayed in CSF of patients with suspected dementia and/or patients with
mild cognitive impairment (MCI) for which is also available a clinical
follow up by means of neuropsychological tests.Techniques available in
the Laboratory of Neurochemistry include mainly immunoassays such as
ELISA and xMAP technique. The latter allows us to multiplex several
analytes increasing the number of molecules that can be measured
starting from the same volume of sample.
CSF biomarkers show promise as a diagnostic tool in patients with AD, but
recent multicenter studies showed that the levels of these biomarkers
vary between different research centers. To study the source of variation
our laboratory is actually involved in a world quality control program run
by the Alzheimer Association in conjunction with Clinical Neurochemistry
Laboratory in Gothenburg, Sweden, with the final goal of standardize
these assays for clinical routine.
Beside the classical CSF biomarkers we are actually studying other
proteins as possible biomarkers for neurodegenerative diseases. We have
recently shown that heart-fatty acid binding protein (H-FABP) is
significantly increased in MCI patients which converted to AD and it may
represent a new marker of neurodegeneration. Other projects are related
to progranulin measurement in CSF, a protein which has been related to
frontotemporal dementia and amyotrophic lateral sclerosis and to the
evaluation of Aβ1-40 as a useful marker to differentiate various dementia
conditions. Finally we are also evaluating the usefulness of the
measurement of α-synuclein and classical CSF biomarkers to distinguish
synucleinopathies (PD and dementia with Lewy Bodies) from other
neurodegenerative diseases.
Finanziamenti ricevuti
Titolo progetto
cNEUPRO (contract no. 199 LSHM-CT-2007-037950)
Ente finanziatore
EU – FP6
Durata progetto
Three years (2007-2010)
Abstract del progetto
Ente finanziatore
Recent research has clearly demonstrated that multiparametric
neurochemical dementia diagnostics (NDD) in cerebrospinal fluid (CSF)
does improve the early and differential diagnosis of dementias. cNEUPRO
will apply advanced proteomic tools to discover novel neurochemical
dementia markers in blood and CSF for the improved early and possibly
predictive diagnosis of AD. A predictive dementia diagnosis will support
the most effective use of forthcoming preventive therapeutic strategies.
Our initiative will establish European SOPs for current neurochemical
dementia diagnostics (NDD). A strong methodological impact is put on
the quality of pre-analytical sample handling and clinical phenotyping,
which has been neglected in industry-driven discovery studies. cNEUPRO
integrates innovative biotech and bioinformatic companies with leading
clinical and proteomic dementia research centres. cNEUPRO will also
support the discovery of new diagnostic targets and is also promising to
identify novel scaffolds for advanced molecular neuroimaging.
CSF lysosomal hydrolases’ activity as possible marker of Parkinson’s
disease
MJFF
Durata progetto
1 year
Abstract del progetto
Objective/Rationale:
Recent studies have linked lysosomal dysfunction to the accumulation of
α-synuclein oligomers and α-synuclein-mediated cell death. Clinical,
neuropathological and genetic associations between Gaucher’s disease, a
lysosomal storage disease, and Parkison’s disease (PD) have been
reported. Also, a reduction of the activities of β-glucocerebrosidase, α
and β-mannosidase has been reported in CSF of PD patients.
To date there is no accepted diagnostic test for Parkinson’s disease based
on biochemical analysis of blood or cerebrospinal fluid (CSF). The
potential use of CSF lysosomal enzyme activities as a diagnostic indicator,
or as a marker of disease progression in PD, will be investigated.
Moreover the CSF levels of alpha-synuclein – a candidate diagnostic
biomarker of PD – will be combined with the lysosomal enzyme activities
in order to verify if there is any association between these biochemical
parameters.Project Description: De-novo and treated patients referring
to the Section of Neurology, University of Perugia, Italy, will be included
in the study. CSF samples will be collected from patients with PD (n=80)
and healthy subjects (n=50). Lysosomal enzyme activities (betaglucocerebrosidase, alpha-mannosidase, beta-mannosidase, beta-
Titolo progetto
hexosaminidase, beta-galactosidase, alpha-fucosidase, arylsulfatase A,
arylsulfatase B, cathepsin D, and cathepsin S) will be determined in CSF
samples using specific fluorimetric substrates. The levels of monomeric
and oligomeric alpha-synuclein, using a sensitive and specific ELISA
method able to reveal levels as low as 1 pg/mL in human biological fluids,
including plasma and CSF, will be also evaluated.
3. HUMAN/CLINICAL RESEARCH
D. Diagnosis: biomarkers
Nome
Contatti
Istituto/Dipartimento
Roberta Ghidoni, Giuliano Binetti
+39-030-3501725
[email protected]
IRCCS “Centro San Giovanni di Dio-Fatebenefratelli”
Proteomics Unit, NeuroioGenLab Memory Clinic, Brescia, Italy
Proposta di ricerca
Over the past 10 years, frontotemporal lobar degeneration (FTLD) has emerged as the most common cause
of dementia under the age of 60 years, outstripping even Alzheimer disease in prevalence. The increasingly
common occurrence of FTLD, and its devasting impact on families and patients within their most
productive years, urges novel approaches to cure or to prevent it. To date, there are no Food and Drug
Administration approved medications for FTLD. FTLD has a strong genetic component, with up to 50% of
cases being caused by mutation. Recently the progranulin gene (PGRN) was shown to be the most frequent
genetic determinant: 68 mutations have been described in 212 families which account for 5–10% of FTLD
cases worldwide (http://www.molgen.ua.ac.be/FTDmutations/). At present, the most common PGRN
mutations worldwide are the Arg493X mutation, which has been identified in 37 patients belonging to 30
genealogically unrelated families with FTLD (the “Mayo Clinic” cohort) (Rademakers R et al,Lancet Neurol.
2007) and the PGRN Leu271LeufsX10 mutation, that we identified in 49 patients belonging to 38 unrelated
Italian families (the “Brescia” cohort) (Benussi et al. NBA 2008;Benussi L and Ghidoni R et al, NBD 2009;
Benussi L et al. ADAD in press).We recently reported that low plasma progranulin levels predict progranulin
mutations in frontotemporal lobar degeneration. (Ghidoni R et al, Neurology 2008). Progranulin therefore
has become a promising target for new therapeutic FTLD approaches. Our resources are:
1) Clinical expertise in FTLD: the IRCCS in Brescia has become a reference center for FTLD and related
disorders serving a large geographical area in Lombardia. 2) The largest (along with the “Mayo Clinic
cohort”) biological sample collection of PGRN mutation carriers (the “Brescia” cohort). Our scientific work
in the last years has generated one of the largest cohort of FTLD families and biological sample collection of
PGRN mutation carriers (affected and presymptomatic), that is an invaluable resource for research studies
3) Cellular and animal disease models.
Our specific aims are:
1) Elucidating the role of progranulin in the disease onset and progression: we will study the molecular
mechanisms underlying neurodegeneration in PGRN mutations carriers (plasma/serum) as well as in
cellular (human fibroblasts and lymphocytes ) and animal models.
2) Biomarkers discovery for the differential diagnosis of FTLD vs Alzheimer’s disease (AD). FTLD is often
misdiagnosed as another type of dementia (AD) and hence biochemical diagnostic markers would aid in
differential diagnosis. Analysis of CSF is so far the most convenient method for studying the biology of
neurodegenerative diseases in living patients. For a global proteomic study, surface-enhanced laser
desorption/ionization mass spectrometry (SELDI-TOF MS) as well as two-dimensional gel electrophoresis
(2-DE) will be used to study differential CSF protein expression.
3) Searching for new genes: Identification of novel disease-associated loci by linkage and gene expression
analysis.
Centers collaborating to this project: IRCCS Centro S.Giovanni di Dio-Fatebenefratelli, Brescia;. Istituto
di Ricerche Farmacologiche "Mario Negri" , Milan (G. Forloni); Università degli Studi di Brescia (PF
Spano); University of Milan (D. Galimberti, E. Scarpini); IRCCS Besta, Milan (F. Tagliavini)
Area di interesse identificata Translational research
Finanziamenti ricevuti
Titolo progetto
Proteomics of cognitive and movement disorders: Identification of
molecular mechanisms associated with disease onset and phenotypic
variability of frontotemporal lobar degeneration- (2009-2633)
Ente finanziatore
FONDAZIONE CARIPLO, Bando Ricerca Biomedica 2009
Durata progetto
01-01-2010/01-07-2012
Abstract del progetto
The main objective of the present study is to unravel the
pathophysiological bases of progranulopathies by
1) proteomic approach- For a global proteomic study, SELDI-TOF as well
as two-dimensional gel electrophoresis (2-DE) will be used to study
differential protein expression in subjects carrying or not PGRN mutations
2) neuroimaging study- The aim of the neuroimaging study is to describe
the MR features of PGRN mutations with conventional, non-conventional,
and functional neuroimaging.
3) personality and behavioral assessment- We will assess the personality
and behavioral trait of asymptomatic at risk family members carrying
PGRN mutations
4) cellular and animal models studies-
Titolo progetto
Project 1: “Validation of genetic and biochemical markers for early
diagnosis of AD and the prediction of conversion of MCI into AD, and
design of a multivariate molecular protocol having high diagnostic and
prognostic accuracy”.
Project 2: “Marker biologici di neurodegenerazione utilizzabili nella
pratica clinica ai fini della diagnosi precoce e differenziale, della
individuazione delle forme con risposta ottimale alle attuali terapie”
Project 1: Italian Ministry of Health, Progr. Strategico PS39, prog. 1
Project 2: Italian Ministry of Health, Progr. Strategico conv.71, prog. 6
Project 1: 01-01-2009/31-12-2010
Project 2: 07-01-2008/07-04-2010
These programs have two primary aims:
1) The development and validation of a multi-factorial protocol that
integrates molecular, imaging, neurophysiological, neuropsychological
and behavioral data for early diagnosis of AD, in particular during the
preclinical phase and the prodromal stage of "mild cognitive impairment"
when the symptoms are not severe enough to fulfill the current
diagnostic criteria for AD.
2) Validation of this new diagnostic protocol within a regional public
health network, and evaluation of the health care, organization and
economic implications of its transfer to the National Health Service.
Ente finanziatore
Durata progetto
Abstract del progetto
Referente: Prof. Carlo Ferrarese
Phone. 039.233.3595
Fax. 039.233.2449
Email: [email protected]
Clinica Neurologica, Ospedale San Gerardo
Via Pergolesi 33 – 20052 Monza (MB)
Laboratorio di Neurobiologia
Dipartimento di Neuroscienze e Tecnologie Biomediche – Università di Milano-Bicocca
Via Cadore 48 – 20052 Monza (MB)
The main interest of our group consists in the integration of clinical and basic scientific research to
understand the pathogenic mechanisms of neurodegenerative disorders and to identify new strategies for
diagnosing, treating and preventing such untreatable diseases. Our center is composed by a clinical core
(Department of Neurology, 42 beds plus Laboratory of Neurophysiology and Laboratory of
Neuropsychology) at the san Gerardo Hospital (Monza, Italy) and a biological facility (Laboratory of
Neurobiology, Department of Neuroscience and Biomedical Technologies, University of Milano-Bicocca).
Regarding dementias and cognitive dysfunction, the main body of our clinical work is centered at the
Memory Clinic (U.V.A. outpatient facility) that sees more than 300 patients among Alzheimer’s disease (AD)
and other types of dementia. Clinical activity is deeply involved with research epidemiological, clinical, and
neurobiological. Novel experimental therapies are offered; for example, a phase 3 trial testing the efficacy
of an anti-beta amyloid antibody in AD is currently recruiting patients.
The strict connection existing between the Memory Clinic and the Laboratory of Neurobiology allows
studying novel peripheral markers in AD (mainly biochemical alterations in blood elements and skin
fibroblasts) that might in the future be used for accelerating diagnostic processes, helping during follow-up,
targeting personalized therapies, and refining prognostic previsions. Bioresearch methods currently
employed include: cell cultures, immunochemistry, molecular biology, binding and uptake techniques,
HPLC, confocal microscopy. For example, our group recently demonstrated an increase of sAPPαplasma
levels with respect to healthy controls, while no differences were found regarding beta-amyloid (Abeta)
levels. Moreover, we set up an ELISA assay for the determination of plasma anti-Abeta 1-42 levels, although
no differences were found between AD patients and controls. However, we observed that AD patients not
receiving acetylcholinesterase medications (AChEI) displayed anti-Abeta 1-42 antibodies plasma levels
significantly lower with respect both, healthy controls, and AD patients receiving AChEI. Hence, we are
currently analyzing the effect of the AChEI therapy on the immune response in AD patients. A putative
immunomodulatory effect of these drugs might eventually further support immunoglobulin-based
therapies for treating dementia. Furthermore, our group extensively studies the interactions existing
between glutamatergic system, APP processing and signal transduction mechanisms (kinases) in peripheral
(platelets, fibroblasts, lymphomonocytes) and cell models (neuroblastoma SHSY5Y). Another
neurobiological interest of our group is related to Abeta catabolism. We focused on neprilisin, a major
Abeta catabolic enzyme, studying its activity and expression in human fibroblasts. Neprilisin activity is
reduced of ~50% in AD versus controls, while the expression in unchanged. In vitro “aged” Abeta treatment
reduces neprilisin expression in both groups, while the activity decreases in controls and increases in AD
patients, possibly due to a compensatory mechanism; this preliminary data suggests that Abeta is able to
influence its own catabolism. A more innovative approach for reducing the toxic effect of Abeta consists in
administering nanoparticles, molecules of nanometric dimensions able to bind Abeta limiting its
aggregation and resulting toxicity. Within an FP7 European project we are testing the biocompatibility of
various nanoparticles, and their ability to bind Abeta in vitro (cultured fibroblasts) and in biological fluids
(plasma and serum) in order to identify the best candidates for future in vivo administrations.
In the neuropsychological field, our current interests include: (a) development and validation of multiple
ultrafast tests (5 minutes duration max) of the global cognitive status in order to propose them as rapid
screening tools for general practitioners; (b) validation of a test of divided attention for the differential
diagnosis of degenerative dementias; (c) development and validation of a novel test of denomination for
the differential diagnosis between mild cognitive impairment (MCI) and dementia; (d) extensive study of
visuo-spatial functions by a broad test battery, in order to define the differential cognitive profile between
Lewy body dementia, corticobasal degeneration and posterior cortical atrophy; (e) study of the
experimental neuropsychological (decision making, moral judgment, multi-tasking, humour appreciation)
and neurobiological correlates of BDNF in AD, frontotemporal dementia and Lewy body or Parkinson
dementia; (f) multicentric longitudinal cohort study recruiting patients affected by degenerative and
vascular dementias or MCI to be followed for five years by repeatedly administering a neuropsychological
test battery.
Finally, our clinical department collaborates with the Department of Nuclear Medicine - Molecular
Bioimaging Centre for both clinical and research purposes. Currently we are studying the existence of
correlations between biochemical markers and functional neuroimaging data (PET-SPET). Tracers able to
mark beta-amyloid or microglial activation, among others, might eventually integrate and confirm the value
of the above described peripheral markers.
Nome
Piero Parchi M.D., Ph.D.
Contatti
[email protected]
phone: 051-2092740
fax: 051-2092751
Istituto/Dipartimento
Dipartimento di Scienze Neurologiche
Università di Bologna, Italy
Proposta di ricerca
Aree di interesse identificate
-
Biobanking (blood sample, CSF, brain repository…)
Standardization of diagnostic criteria and diagnostic
instruments, harmonization and assessment tools
Early diagnosis; Biomarkers
Basic research: Biochemical characterization of
abnormal protein aggregates to understand phenotypic
diversity in neurodegenerative diseases
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Durata progetto
Abstract (estratti) del
progetto
1) Molecular Pathology of the Human Brain;
2) Development of new diagnostic approaches for prion
diseases
3) Validation and search of CSF biomarkers for the prediction
of the clinical conversion from mild cognitive impairment
(MCI) to dementia
1) EU, 2) Italian Ministry of Health, 3) Galletti Foundation
1) 5 years (Brain-Net II, project ended December 2009), COST
application currently under evaluation (at final step).
2) 3 years
3) 3 years
1) Studies on human brain tissue have enabled the discovery
of key factors of the most common brain diseases. Aβ, tau,
synuclein and TDP-43 are outstanding examples of proteins
identified in diseases such as Alzheimer (AD), Parkinson (PD)
and Frontotemporal Lobar Degeneration (FTLD). The advent of
molecular genetic techniques enabling insights into temporal or
permanent changes of genetic activity (epigenetics) and the
identification of novel regulators of translation (miRNA) have
ushered in a new era that makes investigations of human brain
tissue still indispensable. In this project ) we will provide highquality human brain tissue as an essential research resource by
applying and further developing a shared database, common
diagnostic criteria and ethical standards at European level.
Furthermore, new technologies will be employed to investigate
the causes of neurodegenerative disorders using such tissues.
This collaborative group of experts (see http://www.brainneteurope.org) competent in all aspects of work with human brain
tissue will foster European research on these devastating brain
diseases.
2) The overall objective of this project is to improve the
diagnosis and the prognosis of human TSE diseases through the
development of new biochemical tests and the optimization of
already available diagnostic tools. To this aims we will apply
new proteomic approaches to identify TSE-specific patterns in
the CSF and in plasma (or serum) of patients with sporadic CJD
and we will characterize novel biochemical properties of the
abnormal PrPTSE aggregates that may further improve the
strain-specific diagnosis of the disease subtype.
3) As one of the Units involved in a larger project (see Form
sent by Dr. Gallassi from our Department) we will validate and
search for molecular CSF markers that might be of value in
predicting the conversion from MCI to full-blown dementia and
critical for the monitoring of patients once new therapies
become available. We will focus on already established markers
such as A-beta, total tau and fosfo-tau and more recently
discovered molecules such as TDP-43 and progranulin. Different
techniques such as Elisa and western blotting will be applied.
The data will be correlated with those obtained from the
neuroimaging and neuropsychological serial evaluation.
SICILIANO
Analisi biochimica di parametri di stress ossidativo.
Uno dei problemi clinici più attuali riguardala difficoltà a comprendere se lievi disturbi della memoria in
età senile siano oppure no segni iniziali di Malattia di Alzheimer. A tal proposito va ricordato come
attualmente si faccia riferimento a un quadro clinico-neurologico quale il decadimento cognitivo lieve
(“Mild Cognitive Impairment o MCI”) ,quale condizione caratterizzata, come appunto il termine indica, da
una lieve compromissione di funzioni mentali, in primo luogo la memoria, ma potenzialmente a rischio di
sviluppare nel tempo una malattia di Alzheimer e pertanto da seguire attentamente nel suo ulteriore
decorso. Un altro aspetto fondamentale è inoltre rappresentato dal fatto che, in assenza di un marcatore
biologico in vivo che permetta di fare diagnosi di certezza della malattia in vita, assume particolare
importanza la ricerca di parametri di laboratorio che siano in grado, in aggiunta a fornire elementi
indicativi sulle cause di malattia, di essere indicatori di eventuale rischio, evoluzione e diagnosi precoce
di malattia, oltre che predittivi di risposta al trattamento.
Lo stress ossidativo è stato chiamato in causa come fattore implicato nella patogenesi delle malattie
neurodegenerative, inclusa la malattia di Alzheimer, in base all’osservazione che si ha evidenza precoce
di danno mediato da specie reattive dell’ossigeno, non soltanto a livello del sistema nervoso centrale ma
anche a livello periferico.
Poiché le specie reattive dell’ossigeno sono molecole altamente instabili, la loro misura diretta in
campioni di plasma o siero può non riflettere la loro reale concentrazione in vivo; per tale motivo
vengono dosati solo i prodotti finali di stress ossidativo. Distinguiamo i biomarker possono essere distinti
in due grandi gruppi:
1.
Marker di danno ossidativo alle proteine, ai lipidi e al DNA, che rivelano lo stato di ossidazione di
tali macromolecole e che danno un’idea del grado di gravità del processo degenerativo in atto all’interno
della cellula;
2.
Marker dello stato antiossidante, che riflettono la capacità totale dei sistemi cellulari, enzimatici
e non enzimatici, di difesa contro l’attacco delle specie chimiche reattive.
Presso il nostro laboratorio sono valutati markers di danno ossidativo alle proteine (AOPP), markers dello
stato antiossidante quali la capacità ferro-riducente del plasma (FRAP) e la concentrazione del glutatione
totale, ridotto e ossidato.
Gli AOPP consistono in un insieme di proteine tra cui la tiroglobulina, la γ-globulina, l’albumina e la
mioglobulina (Witko-Sarsat et al., 1996).
L’elettroforesi delle proteine mostra che il picco di AOPP ad alto peso molecolare è prevalentemente
dovuto all’albumina che appare sotto forma di aggregati che probabilmente derivano da ponti disolfuro
e/o da "cross-linking" della tirosina; al contrario il picco di AOPP a basso peso molecolare contiene
albumina nella sua forma monomerica (Witko-Sarsat et al., 1996). In "vivo" i livelli plasmatici di AOPP
correlano con i livelli di dimeri di tirosina, un marcatore di danno ossidativo delle proteine, e con la
pentossidina, un prodotto di glicosilazione strettamente associato al danno ossidativo delle proteine
(Selmeci et al., 2005). La relazione tra gli AOPP e i prodotti di perossidazione lipidica appare invece poco
chiara. Le attuali conoscenze sembrano indicare che i lipidi non sono necessari per la formazione degli
AOPP, ma che in “vivo” possono aumentare tale processo (Witko-Sarsat et al., 1996).
Al valore della FRAP contribuiscono, per circa il 60% l’acido urico, per il 15% l’acido ascorbico, per il 5%
l’α-tocoferolo, per il 10% le proteine e per il 5% la bilirubina (Benzie et al., 1996). La metodica che
permette di determinare questo parametro sfrutta la capacità di questi antiossidanti di ridurre il ferro, sotto forma di ione ferrico presente nel reattivo FRAP, - a ione ferroso.
La principale funzione del glutatione è quella di operare come antiossidante proteggendo le cellule
dall'azione di specie proossidanti (Dringer et al., 2000). Le proprietà antiossidanti del glutatione sono
dovute alla presenza del gruppo sulfidrilico della cisteina che gli permette di interagire sia con specie
reattive dell’ossigeno, sia con altre sostanze elettrofile nell’ambito di numerosi sistemi antiossidanti e
non (Pompella et al., 2003). Il glutatione allo stato ridotto (GSH) espleta la sua funzione riducendo i
radicali liberi e convertendosi in glutatione ossidato (GSSG), composto da due molecole di glutatione
unite da un ponte disolfuro. In questo senso, il GSH interviene nel metabolismo dell’acqua ossigenata,
degli idroperossidi e di altri perossidi organici, è capace di inattivare il radicale idrossile, radicali
perossilici, il perossinitrito, l’acido ipocloroso, radicali alcossilici e l’ossigeno singoletto. Può anche
ripristinare la forma ridotta di altri antiossidanti come la vitamina E e la vitamina C.
Valutazione clinica, test da sforzo e prelievi ematici per la valutazione dello stato ossidativo
I parametri di stress ossidativo sopra descritti possono essere studiati sia in condizioni basali che dopo
protocolli di esercizio. Le principali sedi di produzione di radicali liberi sono i mitocondri, gli organelli
intracellulari deputati al metabolismo ossidativo; i mitocondri svolgono altresì un ruolo di primo piano
nel mantenere l’omeostasi del calcio e nell’innescare la cascata di segnali che sottendono la
degenerazione cellulare attraverso il meccanismo della apoptosi.
Lo studio della funzione mitocondriale e dello stress ossidativo ad essa connesso può essere valutato
attraverso l’andamento dei parametri ematici di stress ossidativo in relazione all’esercizio muscolare.
Esempio di test da sforzo per la valutazione di parametri di stress ossidativo:
I pazienti sono sottoposti a un test di sforzo incrementale sui muscoli dell’avambraccio, eseguito con
l’ausilio di un miometro connesso ad un “hand-grip” (Digital Multi-Myometer, MIE Medical Research Ltd.,
Leeds, UK). Il protocollo di esercizio incrementale consiste nell’impugnare l’hand-grip del miometro e
contrarre massimamente la mano contro la resistenza offerta dallo stesso, determinando, in Newton, il
livello di contrazione volontaria massimale (CVM). Saranno eseguite, da tutti i pazienti, una serie di fasi
contrattili o “steps”, condotte in maniera intermittente per un periodo di un minuto, con intervalli di
riposo di 2 minuti tra uno “step” e l’altro. In ciascuno “step” le contrazioni intermittenti sono eseguite con
frequenza di una al secondo contro la resistenza richiesta offerta dal miometro. L’esercizio inizia con un
primo “step” al 10% della CVM, un secondo “step” al 40% della CVM ed un ultimo “step” al 70% della
CVM. Il paziente potrà seguire su un display luminoso il livello di forza generato in ogni contrazione.
Questo tipo di esercizio è principalmente aerobico all’inizio del test e diviene progressivamente
anaerobico man mano che aumenta la forza esercitata per progressivo reclutamento delle unità motorie
rapide (Milner-Brown et al., 1973). I prelievi ematici venosi per la determinazione dei marcatori
biochimici di stress ossidativo sono eseguiti a livello basale, dopo il secondo “step”, dopo il terzo “step” e
dopo 15 minuti dalla fine dell’esercizio.
Nome
Laura Calzà
Contatti
[email protected]
Tel. +39 051 2097947
Cell. +39 335 310979
Istituto/Dipartimento
DIMORFIPA
Proposta di ricerca
Area di interesse identificata
Multidisciplinary projects
Cognitive decline in multiple sclerosis. Cognitive defects are
recognized as early events in MS, not correlated with white matter
lesion. We are interested in studying the pathogenesis of early
neural distress and lesion in MS animal models and neuroprotective
strategies
Standardization of diagnostic
criteria and diagnostic
instruments, harmonization
and assessment tools
Quality control programs for Abeta fragments in CSF and plasma
and tau/Ptau in CSF using xMAP platform. Focus on clinical trials
standardization
Finanziamenti ricevuti
Titolo progetto
Translational medicine for novel diagnostic and therapeutic tools for
neurodegenerative diseases
Ente finanziatore
Joint Regione Emilia Romagna-University of Bologna
Durata progetto
3 years, under negotiation
Abstract del progetto
Development of a GLP-grade service for drug testing in animal
models of Alzheimer disease, based on computerized analysis of
video-tracking for learning and memory performance. Development
Titolo progetto
of cell-based platform based on neural stem cells derived from
Alzheimer mice for the screening of chemical libraries of antiAlzheimer drugs.
Effect of low-dose of ….. in chronic administration on cognitive
behavior and morphological parameters in Tg2576 mouse
Ente finanziatore
Drug Company
(covered by disclosure agreement)
Durata progetto
12 months
Abstract del progetto
The project is aimed to evaluate efficacy of a novel gammasecretase inhibitor on behavioral performance, histopathology,
biomarkers and synapse morphology and molecular markers in mice
curring the swedish mutation of APP. This esperiment will test
efficacy of chronic treatment from 5 to 19 months of age. Two
doses of the test compund and one dose of a reference will be
tested in transgenic vs wild tipe, age matching mice
Plasma assay of Ab40/Ab42 (multiplex X-MAP-based immuno assay
technology) in phase 1 clinical trial
Titolo progetto
Ente finanziatore
Drug Company
(covered by disclosure agreement)
Durata progetto
20 months
Abstract del progetto
Placebo-Controlled, Ascending Single-Dose Study to Evaluate the
Safety, Pharmacokinetics and Pharmacodynamics of …. in Healthy
Young Male Subjects. The safety and tolerability of five ascending
oral doses of … will be evaluated in 6 separate study sessions. In
each of the study sessions, subjects will be randomized to … or
placebo treatment. Overall, 60 subjects will participate in the study,
48 treated with … and 12 treated with placebo. Our lab in
committed in efficacy variables blind analysis (Plasma -amyloid
concentrations, A40/A42 fragments). Joint QC initiative “round
Robin” study, an international interlaboratory evaluation of AB 140/1-42 levels from a common plasma sample set, toward
standardization of multiplex/multiparametric methods for AB 140/1-42 quantification in plasma. Participating centers, 15; number
of samples to analyze, 19.
Remyelination failure in multiple sclerosis: a case of inflammationinduced tissue hypothyroidism?
Titolo progetto
Ente finanziatore
AISM/FISM
Principla investigator: Luciana Giardino
Durata progetto
2 years
Abstract del progetto
Nome
This project is aimed to investigate a possible mechanims for
remyelination failure in multiple sclerosis. This project is aimed to
verify if the inflammation associated to demyelination causes a
defect of T4 to T3 conversion or a decreased expression of thyroid
hormone receptors (nuclear receptors and membrane transporters)
in the nervous tissue, thus leading to a local/cellular hypothyroidism
and, thus, to the defect of white matter repair. This project could
also provide insides for other neurodegenerative disease including
white matter alterations, like Alzheimer disease
Laura Calzà
Contatti
[email protected]
Tel. +39 051 2097947
Cell. +39 335 310979
Istituto/Dipartimento
DIMORFIPA
Proposta di ricerca
Area di interesse identificata
Development of competitive
animal models for AD
GLP grade animal facility and behavioural testing for cognitive
performance in AD animal models using video tracking and
computer analysis; screening of new molecules after acute
administration
Basic research
Brain metabolism of thyroid hormone: a risk factor for AD? We are
interested in exploring possible connections between thyroid
hormone content, receptor and activating enzyme expression, APP
metabolism and Ab toxicity in the brain in normal and AD mice.
Recent advances in knowledge on thyroid hormone brain
metabolism and molecular biology of nuclear and membrane
thyroid hormone receptors and transporters offer new possible
targets to explore this topic.
New treatment strategies
Nome
Contatti
Istituto/Dipartimento
Fabrizio Tagliavini
Tel +39 02 2394 2260; Fax +39 02 2394 2101
Email: [email protected]
Sito web: http://www.istituto-besta.it
UO Neuropatologia – Neurologia 5
Fondazione IRCCS Istituto Neurologico “Carlo Besta”
Via Celoria ,11 - 20133 Milano- ITALY
Proposta di ricerca
The research group: The Division comprises (i) a Clinical Unit (Dementia Center) devoted to the diagnosis
and treatment of patients with degenerative dementias (over 2000 out-patients and 150 in-patients per
year) and (ii) a Laboratory Unit (certified ISO 9001:2000 - Registration Number 26080, dedicated to the
analysis of genes and biomarkers associated with degenerative dementias, post-mortem characterization
of the disease process and disease-specific protein, and experimental studies on disease pathogenesis and
the development of therapeutic strategies. The course of action of this activity can be expressed as a “Bed
to Bench to Bed” cycle, with the ultimate goal to identify disease-modifying drugs in preclinical settings
and apply them to patients. In this regard, the Clinical Unit is currently coordinating a multicentre phase II
clinical trial supported by AIFA, to test the effectiveness of an anti-amyloidogenic molecule identified in
experimental models. This comprehensive approach has also the great advantage to collect biological
samples (plasma, DNA, CSF and brain tissue) from fully characterized patients. In this regard the
Laboratory Unit is part of the Network of Excellence “BrainNet” Europe devoted to biobanking,
harmonization of assessment tools and standardization of diagnostic criteria of neurodegenerative
diseases.
The permanent staff of the Division is composed by 8 MD and two technicians. In addition, the staff
comprises 13 post-doctoral fellows (7 PhD, 3 MDV, 1 MD, 1 psychologist), 2 PhD students and 1 technician
who are committed to the research activities on degenerative dementias with different expertise and
roles. On the overall, the team has large experience in a variety of techniques spanning from
neuropathology (including immunohistochemistry, morphometry, electron microscopy and atomic force
microscopy), to molecular genetics, biochemistry (purification and characterization of disease-specific
proteins), cellular and molecular biology, and animal models. The Division of Neuropathology is provided
with fully equipped laboratories for histology, immunohistochemistry, electron microscopy, atomic force
microscopy, biochemistry, and molecular and cellular biology, and a 60 sqm BL3 facility. Moreover an
animal facility composed by a 140 sqm environmental controlled area, provided with a HVAC system and
heap-filtered ventilated racks, with separate rooms for maintaining and breeding mouse lines, and
carrying out surgical procedures, collection of samples and post-mortem examination is available.
Research lines on Alzheimer’s disease
A. Clinical research
• Early diagnosis of Alzheimer’s disease (AD)
We will characterize the phenotypic expression of AD and other degenerative dementias through
multiplex analysis of a number of variables including neuropsychological and behavioural profile,
neuroimaging, neurophysiological changes with special attention to sleep abnormalities, genetics, and
plasma and CSF biomarkers. Furthermore, we will carry out longitudinal studies of presymptomatic
carriers of gene mutations to detect early markers of conversion to the disease state.
• Clinical trials
We will carry out phase II and phase III clinical trials to assess the effectiveness of potentially diseasemodifying drugs, including molecules targeted to Aβ and protein tau.
• Identification of disease-specific biomarkers
We are currently working on a peripheral marker of AD having high specificity and sensitivity, based on
cyclic amplification of misfolded Aβ and Aβ oligomers from biological fluids, similar to the PMCA technique
successfully developed by Claudio Soto for prion diseases. The same approach will be used to amplify and
detect misfolded tau.
• Biobanking and neuropathological and molecular characterization of patients
We will collect systematically plasma, DNA and CSF from patients with different types of dementia and
non-demented individuals, and perform autopsies for neuropathological characterization and biochemical
and molecular studies.
B. Preclinical research
• Recessive A673V APP mutation: molecular mechanisms and development of a new therapeutic strategy
for sporadic AD
We have recently identified an APP mutation (A673V) that causes early-onset AD only in the homozygous
state while the heterozygous carriers are not affected. This mutation strongly boosts the production and
amyloidogenic properties of Aβ. However, the interaction of A673V-mutated and wild-type peptides
inhibits amyloidogenesis and A-mediated neurotoxicity (Di Fede et al., Science 2009, 323:1473-7). These
findings are consistent with the observation that the A673V heterozygous carriers do not develop disease
and offer grounds for a novel therapeutic strategy based on modified Aβ peptides. A research priority of
our lab is to unravel the molecular mechanisms of the opposite effects of the A673V APP mutation in
homo- or heterozygous state on amyloidogenesis and to develop a lead compound for AD therapy based
on A673V-modified Aβ peptides or peptido-mimetic molecules. To accomplish these objectives we have
generated a panel of transfected cells and transgenic C. elegans expressing human APP or Aβ with the
A673V mutation, respectively, and transgenic mouse lines expressing A673V-mutated APP in the
homozygous or heterozygous state. Furthermore, we have identified a prototypic lead compound
corresponding to a six-mer Aβ peptide with the A673V substitution and are currently working on brain
delivery systems.
• Pathways leading to tau pathology
A key event in AD pathogenesis is the hyperphosphorylation, misfolding and aggregation of the
microtubule-associated protein tau leading to formation of neurofibrillary tangles, disruption of the
neuronal cytoskeleton and neurodegeneration. Following the “Aβ cascade hypothesis” this event is
triggered by aggregated Aβ species, particularly oligomeric assembly intermediates. However, the
pathways linking Aβ and tau pathology are unknown. This is largely due to lack of animal models able to
reproduce these two central aspects of AD. We have developed a mouse model of prion disease that
shows a secondary tauopathy following deposition of PrP amyloid (Giaccone et al., Neurobiol. Aging 2008,
29:1864-73). This model will be used to investigate the molecular basis of tau pathology induced by
deposition of an amyloid protein.
• Role of nuclear tau in neurodegeneration in fronto-temporal dementia
Tau is the major microtubule-associated protein of neurons and its primary role is to promote assembly
and stabilization of microtubules required for morphogenesis and axonal transport. We have recently
found that tau plays an important role also in chromosome stability, and mutations in the tau gene cause
chromosome aberrations in peripheral cells in addition to formation of neurofibrillary tangles in neurons
and glial cells. Our objective is to elucidate the molecular mechanisms underlying the genomic instability
due to tau mutations using cellular models, and to investigate the contribution of the aneuploidy caused
by mutated tau to neurodegeneration using a transgenic mouse model of tauopathy.
Area di interesse identificata - Genetic susceptibility to Alzheimer’s disease (AD)
CLINICA/PRECLINICA
- Early diagnosis
- Biomarkers
- Developing competitive animal models to study AD
- Studying early onset forms of AD to follow-up disease progression
- Cinical trials with or without involvement of pharmaceutical companies
- Basic research
- Biobanking (blood sample, CSF, brain repository…)
- Standardization of diagnostic criteria
- New treatment strategies
- Translational research
Finanziamenti ricevuti
1. Titolo progetto
Genoproteomics of Age Related Disorders (GuARD).
Ente finanziatore
Durata progetto
Abstract del progetto
2. Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
3. Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
4. Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
5. Titolo progetto
Ente finanziatore
Durata progetto
Abstract del progetto
PI: Roberto Sitia Subproject leader: Fabrizio Tagliavini
Cariplo Foundation
01/01/07-30/06/10
Project 3.2: Neurodegeneration. The main goals of this project are: (1)
biochemical analysis of misfolded protein aggregates from brains of
patients with sporadic and genetic forms of Alzheimer disease and prion
diseases; (2) proteome analysis on brain tissue from the same patients
and from gender- and age-matched non-demented control individuals; (3)
proteome analysis on brain tissue from a mouse model of variant
Creutzfeldt-Jakob disease.
Alzheimer's disease: development and validation of a multi-factorial
protocol for the diagnosis and follow-up of disease in the prodromal and
incipient phase. PI: Fabrizio Tagliavini
Italian Ministry of Health
01/01/09-30/06/11
This program has two primary aims: (1) the development and validation
of a multi-factorial protocol that integrates molecular, imaging,
neurophysiological, neuropsychological and behavioral data for early
diagnosis of AD, in particular during the stage of MCI; (2) the validation of
this diagnostic protocol within a public health network, and evaluation of
the health-care, organization and economic implications of its transfer to
the NHS.
Molecular mechanisms underlying synaptic dysfunction in prototypic
neurodegenerative diseases related to protein misfolding (nEUROsyn). PI:
Fabrizio Tagliavini
European Union/Italian Ministry of Health
01/02/09-31/01/12
This project is designed to investigate the mechanisms that lead to
synaptic impairment and demise of neurons in AD and Huntington
diseases using cellular and animal models, and patient material. The main
goals are to: (1) study synaptic dysfunction; (2) identify alterations of
synaptic and dendritic spine remodelling; (3) monitor changes in receptor
trafficking at the synapse and disturbances in trafficking of different
cellular components.
A randomized, double-blind pilot study versus placebo for the evaluation
of the efficacy of doxycycline administered by oral route in patients
affected by Creutzfeld-Jakob disease. PI: Fabrizio Tagliavini
Italian Italian Agency of Drug (AIFA)
01/09/06-31/08/10
This project is a multicentre phase II clinical trial to test the effectiveness
of an anti-amyloidogenic molecule (doxycline) identified in experimental
models.
Biological markers of neurodegeneration to be used in clinical practice for
early and differential diagnosis of degenerative dementias, the
identification of fast and slow progressing forms, and the evaluation of
response to treatment. PI: Gianfranco Spalletta, Project Leader: Fabrizio
Tagliavini
Italian Ministry of Health
01/07/08 to 01/06/10
The main goal of the project is the identification of peripheral markers of
neurodegeneration in Alzheimer disease, Levy body dementia,
frontotemporal dementia and prion diseases to allow (1) early differential
diagnosis, (2) identification of fast and slow progressing forms, (3)
evaluation of response to therapy.
Nome
Contatti
Tel.
E mail
Istituto/Dipartimento
Gianfranco Spalletta
Via Ardeatina, 306 – 00179 Rome
06-51501575
[email protected]
IRCCS Santa Lucia Foundation/Department of Clinical and behavioral
neurology - Neuropsychiatry laboratory
Proposta di ricerca
Area di interesse identificata
Mild Cognitive Impairment
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Behavioral and Psychological Predictors of Cognitive Outcome in Subjects
with Mild Cognitive Impairment: the Role of Non-conventional
Neuroimaging and Serotonergic Genes
Italian Ministry of Health
Durata progetto
01/09/2010 – 31/08/2012
Abstract del progetto
The present project will identify complex behavioural-endophenotypes,
of behavioral-neuroimaging-genetics nature, useful for predicting the
cognitive course of neurodegenerative dementias starting from their
preclinical phase or MCI. In particular results will help to predict the
outcome of the cognitive impairment, the relationship between cortical
and subcortical brain atrophy and neuropsychiatric-neuropsychological
longitudinal clinical manifestations, and a composite neuroimagingserotonin genetics index for better prediction of behavioral expression
during the 18 months longitudinal course of the project. In order to do
this an effort will be made to try to make the predictive capability of each
clinical marker reliable for every single patient.
Two-hundred people with a diagnosis of MCI will be recruited in this
study. These patients will be drug free from psychopharmacological and
acetilcholinesterase inhibitor drugs. One-hundred healthy controls (HC)
will be also recruited. The subjects included will carry out an in depth
anamnesis evaluation, a diagnostic evaluation for depression, psychosis,
and apathy associated with dementia, and a psycho-behavioural
dimensional assessment using a battery of tests including:
Neuropsychiatric Inventory; CERAD Disforia; Dementia Apathy Interview
Rating, Apathy Scale and Pittsburgh Sleep Quality Index. Moreover, the
subjects will be evaluated through the MMSE, the Mental Deterioration
Battery (MDB) and tests for basic and instrumental functional activities.
This is a mixed case-control study (MCI at the baseline vs. HC) and
longitudinal study (18-month follow-up only for MCI subjects). The first
six months of research will be used for the inclusion of HC and MCI
subjects in the study. Each cognitive and behavioural battery tests and
diagnostic criteria will be administered in the MCI subjects every six
months until 18 months from the first evaluation. Caregiver stress will be
monitored at the baseline and the end of the follow-up. Image acquisition
will be done at the baseline: all subjects will be examined using a 3 Tesla
Allegra MR Imager with a standard quadrature head coil. Participants will
undergo the same MR imaging protocol including whole-brain T2*weighted, T1-weighted and DTI scanning. Standard clinical sequences
(FLAIR and T2-weighted images) will be acquired on the same subjects in
order to exclude different pathologies. Image processing will be
performed using FreeSurfer, FSL 4.1 and in-house developed software in
Matlab vers. 6.5. The FreeSurfer toolkit will be employed to perform
automatic cortical thickness reconstruction. The high resolution
anatomical T1-weighted images will be also used to extract several region
of interest corresponding to subcortical grey matter nuclei. The automatic
segmentation of these nuclei will be obtained with the software FIRST,
part of the FSL toolkit. Moreover, the DTI data will be coregistered to the
T1 images in order to extract regional values of diffusion parameters (MD,
FA) known to be indicative of tissue microstructure disruption, from the
same nuclei identified in the previous step. The DTI data will be also
employed to identify the main fiber tracts within the white matter. These
tracts that will represent additional regions of interest from where
extracting diffusion parameters. Genomic DNA will be purified from 200
microl of human whole blood. Polymorphisms and haplotypes of several
genes will be analyzed in this study: the Apolipoprotein epsilon 4 (ApoE4)
variant will be investigated in HC and MCI subjects in order to verify its
influence on behavioural manifestations and symptom progression; the
102T/C polymorphisms of the 5HT2A receptor gene will be investigated
for psychosis behavioural phenotype; to test the hypothesis that allelic
variation in 5HTT gene-linked polymorphic region (5-HTTLPR) genotype is
associated with behavioural syndromes and symptoms and cognitive
outcome, a common 44-base pair deletion (s allele) polymorphism in the
5-HTTLPR which associated with reduced 5HTT transcription efficiency
and 5HT uptake in vitro will be studied.
The main focus of the research in the field of complex disorders, such as
dementias, is to found new complex predictors which are valid and
reliable in the prediction of clinical outcome starting from the early
preclinical phases. In particular, mixed clinical (behavioural) and biological
(neuroimaging, genetics) predictors may help to precociously identify
those subjects who have a rapid decline of cognitive performances and
those who will respond positively to the treatment with a stable course of
the cognitive performances and who also improve. Another interesting
possibility is that exist behavioural variables which may predict the course
of the illness in the case they will not improve at the end of the follow-up.
The longitudinal branch of this project is devoted to identify if this
possibility exists and which are the behavioural predictors in terms of
frequency of behavioural disorders at the end of the follow-ups.
3. HUMAN/CLINICAL RESEARCH
E. Diagnosis: neuropsychology
Nome
Contatti
Istituto/Dipartimento
Maurizio Balestrino
010-3537078/7085/7066; [email protected]; FAX 0103538631; mobile phone 348-7814154
Department of Neuroscience, Ophtalmology and Genetics, University of
Genova, Italy
Proposta di ricerca
Dementia of Alzheimer's type represents about 40% of the cases where a last will is challenged based on
real or supposed mental incapacity of the testator (1). Usually, elder people find in their last will the
reassurance that after their death their loved ones will have the resources to carry on their lives in
agreement with the old person’s desires. However, when a person suffers (or is suspected to suffer) from
Alzheimer’s disease, their last will is often disputed, on the ground that he/she may not have been
mentally competent to make decisions. Thus, the peace of mind that comes with having written a
thoughtful last will is considerably decreased. This is a growing problem, that is already painful from the
social point of view (see for example http://elder-abuse-cyberray.blogspot.com/2007/06/where-there-iswill.html). Moreover, somebody may really take advantage of the declining cognitive status of an
Alzheimer’s patient to have him/her write a will in his/her favor (2). The dispossessed heirs then claim that
the will is invalid because the deceased was, at the time of writing it, incapacitated due to mental
deterioration (3).
In such cases, a judiciary trial follows, however the judgement is often difficult, because the writer of the
will can no longer be examined, and the judgement must rely solely on the retrospective evaluation of
medical records and of witnesses’ statements. Since both types of evidence are often incomplete or even
discordant, the expert’s judgement is often difficult and flawed by obvious limitations. However, the last
will has often been handwritten. If so, the quality of the handwriting may add very important elements to
the expert’s evaluation, because it shifts the investigation from the retrospective medical records or
witnesses’ claims to the direct observation of the deceased subject’s performance. The act of writing is by
no means a mere movement of the hand and fingers, rather it is a cognitive performance from several
viewpoints. The writer must (1) correctly build in his/her mind a suitable sentence, (2) translate words into
graphic symbols, and (3) correctly arrange the writing in rows and columns, in such a way that it can be
decoded. The first two tasks relate to speech abilities, and are usually negatively affected by aphasia. The
third task implies a correct visuospatial ability. Both aphasia and visuospatial disturbances are prominent
symptoms of Alzheimer’s type dementia (4). Accordingly, it has been shown that handwriting is
compromised in demented patients (5,6,7,8). Even more important, a correlation exists between severity
of handwriting disruption and severity of cognitive deficiency (for example, see Table 3 of reference (7)
and Table 7 of reference (9)).
While a wrong or confused handwriting is an obvious sign of mental impairment, the evaluation of this
item in forensic psychiatry is currently hampered by the lack of the objective and quantitative tools that
are required for a forensic examination. Although it has been demonstrated by several authors that
writing is altered in demented patients (7,5,8,6), these data are difficult to apply to the evaluation of a
single case, as it is required for judiciary purposes. By contrast, a quantitative score of handwriting that
was able to identify severe mental deterioration with a high probability level would be an invaluable tool
in the forensic evaluation. In preliminary research we addressed this issue by designing a semiquantitative
standardized score system to evaluate handwriting, and investigated both its inter-rater reliability and its
correlation with standard measures of cognitive performance (10). In that preliminary research we showed
(a) that our semiquantitative handwriting scoring method has good interrater agreement and (b) that
there is a statistically significant correlation between the degree of cognitive deterioration (as gauged by
the score of both Mini Mental State Examination – MMSE - and of Milan Overall Dementia Assessment –
MODA scale) and the handwriting score (10).
We now propose to continue that research by testing a larger number of patients, to more definitely
quantify the correlation between severity of dementia and handwriting score. Specifically, we plan to (1)
Repeat our investigation on a larger number of patients to hopefully better validate it. (2) Administer to
our patients not only the MMSE and MODA tests, but also the Alzheimer´s disease assessment scale (
ADAS-COG), an internationally accepted tool for the assessment of Alzheimer’s disease. (3) Validate the
test by identifying cutoff values of our handwriting score that reliably identify persons with normal mental
capacity and persons with mild, moderate or severe dementia.
We will test the hypothesis that (a) a writing score lower than a certain value (to be indentified in ou
proposed research) reliably identifies patients whose mental deterioration is minimal or inexistent and (b)
a writing score higher than a certain value (writing score=5 according to our preliminary research, see ref.
10) reliably identifies persons with severe mental deterioration.
We will carry out our research using a protocol similar to our published work (10). We will exclude patients
having dementia not of Alzheimer's type, patients having severe motor deficit in the dominant hand, and
patients having aphasia so severe as to prevent communication altogether.
We believe that our reseach will help to bring justice and peace of mind to Alzheimer's disease victims and
to their families, by providing a method, scientifically rooted in clinical neuropsychology, that will help
preventing abuse and exploitation of Alzheimer’s disease patients through their last will. While this is in
itself an important goal of our research, we also believe that the quantification of the relationship
between Alzheimer’s disease and handwriting will open a new way of testing cognitive impairment in
Alzheimer’s disease, with important fallout to the entire field of neuropsychological testing of Alzheimer’s
disease patients.
References
8) Shulman, K. I., Cohen, C. A., and Hull, I., (2005) Psychiatric issues in retrospective challenges of
testamentary capacity. Int.J Geriatr.Psychiatry 20:63-69.
9) Peer, I. N., (1981) Wills, testamentary capacity and undue influence. Bull.Am Acad Psychiatry Law
9:15-22.
10) Redmond, F. C., (1987) Testamentary capacity. Bull.Am Acad Psychiatry Law 15:247-256.
11) Victor, M. and Ropper, A. H., (2005) Adams and Victor's Principles of Neurology. 8th Ed.:
12) Werner, Perla, Rosenblum, Sara, Bar-On, Gady, Heinik, Jeremia, and Korczyn, Amos, (1-7-2006)
Handwriting Process Variables Discriminating Mild Alzheimer's Disease and Mild Cognitive
Impairment. Journals of Gerontology Series B: Psychological Sciences and Social Sciences 61:228236.
13) Croisile, B., (2005) [Writing, aging and Alzheimer's disease]. Psychol Neuropsychiatr.Vieil. 3:183197.
14) Silveri, Maria Caterina, Corda, Francesca, and Di Nardo, Miriam, (2007) Central and peripheral
aspects of writing disorders in Alzheimer's disease. J Clin Exp Neuropsychol. 29:179-186.
15) Forbes, Katrina E., Shanks, Michael F., and Venneri, Annalena, (1-3-2004) The evolution of
dysgraphia in Alzheimer's disease. Brain Research Bulletin 63:19-24.
16) Luzzatti, Claudio, Laiacona, Marcella, and Agazzi, Daniela, (2003) Multiple patterns of writing
disorders in dementia of the Alzheimer type and their evolution. Neuropsychologia 41:759-772.
17) Fontana P, Dagnino F, Cocito L, Balestrino M. (2008) Handwriting as a gauge of cognitive status: a
novel forensic tool for posthumous evaluation of testamentary capacity. Neurol Sci. 29(4):257-61.
18) Brazzelli, M., Capitani, E., Della Sala, S., Spinnler, H., and Zuffi, M., (1994) A neuropsychological
instrument adding to the description of patients with suspected cortical dementia: the Milan
overall dementia assessment. J Neurol Neurosurg.Psychiatry 57:1510-1517.
19) Folstein, M. F., Robins, L. N., and Helzer, J. E., (1983) The Mini-Mental State Examination. Archives
of General Psychiatry 40:812.
Area di interesse identificata
- Standardization of diagnostic criteria and diagnostic
Finanziamenti ricevuti
Titolo progetto
//
Ente finanziatore
//
Durata progetto
//
instruments, harmonization and assessment tools
Multidisciplinary projects
Responsabile: Angiola Maria Fasanaro (Neurologo)
Collaboratori: Raffaele Rea
(Volontario Neuropsicologo)
Anna Carotenuto
( Volontario Neuropsicologo)
Maila D’Anonio
(Volontario Neuropsicologo)
Luisa Colucci
( Tirocinante Neuropsicologo)
Sabrina Carpi
( Tirocinante Neuropsicologo)
Massimiliano Bosco ( Tirocinante Neuropsicologo)
Ivana Molino
( Tirocinante Neuropsicologo)
Gabriella Finizio
(Tirocinante Neuropsicologo)
Antonio Ziello
( Tirocinante Neuropsicologo)
La Unità Valutativa Alzheimer, da me diretta è attiva dal 2000, e fa parte della Struttura Semplice di
Malattie Involutive Cerebrali, Dipartimento di Neuroscienze, AORN "A. Cardarelli".
Le attività dell’ ambulatorio sono prevalentemente svolte su soggetti geriatrici.
L’esperienza maturata nel corso di 10 anni di attività ha permesso di valutare oltre 900 pazienti , ed
attualmente circa 400 sono in terapia regolare. I dati sono raccolti in cartelle cliniche ad hoc ed archiviati
elettronicamente.
I protocolli seguono le Linee Guida Internazionali e si avvalgano dell’ applicazione di test neuropsicologici
specifici di approfondimento. Particolare attenzione viene dedicata all’ amnesic/non amnesic Mild Cognitive
Impairment e ai fattori vascolari associati alle demenze.
I pazienti sono controllati a distanza di uno, tre e sei mesi dall’inizio della terapia, ed ogni sei mesi
successivamente.
Oltre alle attività diagnostiche e di monitoraggio l’Unità svolge un servizio di counseling diretto ai caregiver
e programmi di riabilitazione diretti a preservare capacità residue.
Attualmente il centro si avvale della collaborazione di 9 neuropsicologi tra volontari e tirocinanti che
costantemente si occupano delle attività ambulatoriali e di ricerca. In quest’ ambito è stata svolta una
collaborazione regolare nel triennio 2005-2007 con L’ Università di Tunisi per un progetto di
implementazione delle metodiche neuropsicologiche.(progetto finanziato dal Ministero degli Affari Esteri.)
Abbiamo una collaborazione stabile con la Cattedra di Neuropsicologia Clinica della Seconda Università
degli Studi di Napoli. e con il Dipartimento di Medicina Sperimentale e Sanità Pubblica dell’Università di
Camerino. E in corso di pubblicazione una ricerca realizzata insieme all’Università Orientale di Napoli e all’
Università La Sapienza di Roma.
I principali progetti di ricerca attualmente attivi
1) Caratteristiche neuropsicologiche differenziali tra pazienti con Demenza fronto-temporale e con
variante frontale della Malattia di Alzheimer.
2) Il contributo del network frontale alla genesi delle allucinazioni in pazienti con malattia di Alzheimer e
con malattia di Parkinson.
3) I fenomeni attrattivi spaziali ed il closing-in
4) Caratteristiche fMRI del Mild Cognitive Impairment Amnesico.
5) Caratteristiche neuropsicologiche delle varianti atipiche della Malattia di Alzheimer.
6) Valutazione cognitiva e monitoraggio degli effetti terapeutici dei trattamenti farmacologici
(Rivastigmina patch, Precursore Colinergico Colina alfoscerato, RO5313534 ) sui sintomi cognitivi e
comportamentali della malattia di Alzheimer e nelle forme con danno vascolare associato.
7)
Analisi dei costi sociali ed economici delle demenze.
8)
Arte visiva come proposta di riabilitazione cognitiva e motivazionale
9) Depressione e Malattia di Alzheimer studio sui correlati gene-ambiente e risposta al trattamento
farmacologico.
D.
Nome
Roberto Gallassi
Contatti
[email protected]
Istituto/Dipartimento
Department of Neurological Sciences, University of Bologna
1) Centre for the Neurological Study of Brain Aging – Gallassi
Roberto
2) Neurogenetic Unit – Carelli Valerio
3) Neuropathology Unit – Parchi Piero
Department of Internal Medicine, Aging and Nephrology, University
of Bologna
1) MR Spectroscopy Unit – Lodi Raffaele
Proposta di ricerca
Area di interesse identificata
Earli diagnosis
Finanziamenti ricevuti
Titolo progetto
Predictors of conversion from MCI to dementia
Ente finanziatore
Fondazione Gino Galletti
Durata progetto
3 years
Abstract del progetto
Background
MCI is a well known clinical condition intermediate between normal
ageing and dementia. It is not completely ascertained if this entity is
a beginning of a dementia or a clinical syndrome which can have
different outcomes. Anyway, the annual conversion rate of MCI to
dementia is variable in different previous studies, ranging from
about 4% to 12% or more. The investigated factors of conversion
taken into account are clinical aspects, neuropsychological tasks,
conventional and functional neuroimaging measures, biological
markers and genetic factors.
Previous neuropsychological studies indicates that MCI may be
characterized by subtle impairment in learning and delayed recall,
particularly in tasks of episodic memory and visuospatial memory, in
executive functions, phonemic and semantic verbal fluency, etc. In
our recent study, we found that a behavioural memory battery,
visual attention, long-term verbal memory tasks and
Neuropsychiatric Inventory caregiver distress and apathy scores,
resulted in independent predictors of conversion to dementia. We
also found that single domain impairment, including memory, has a
minor probability to develop dementia than multiple domain
impairment.
Rationale
We hypothesize that some cognitive functions and
neuropsychological tasks are more sensitive in predicting the
conversion of dementia in relationship with modifications in other
clinical, biochemical, genetic and neuroimaging findings.
Aim
The aims of this project are:
-
-
-
-
to evaluate at baseline demographic features, clinical
characteristics, cognitive and behavioural profile, biochemical
and genetic findings in a sample of MCI patients of amnesic or
not amnesic type and with single or multiple domain
impairment.
to perform clinical and MR neuroimaging longitudinal
observation during three years of the MCI sample, evaluating
the evolution of the single patients, in particular the conversion
to dementia
to correlate with a multi-dimensional approach the relationship
between clinical, cognitive and behavioural findings and the
other methods of investigation such as quantitative MR
neuroimaging measures, haematic and cerebrospinal fluid
biological data and the genetic profile of patients
to evaluated the independent predictors of conversion from
MCI to the various type of dementia
F. Biobanking Bioinformatics
3. HUMAN/CLINICAL RESEARCH
G. Ambient assisted living and socioeconomics
Roberto Monastero, MD, PhD
Lab of Epidemiology and Psychology of Aging and Dementia
Dept of Clinical Neuroscience
University of Palermo
Via La Loggia 1
90129 - Palermo, Italy
Tel. +39-091-6555185
Fax. +39-091-6555113
email. [email protected]
[email protected]
PROGETTO2
Type: Ambient Assisted Living (AAL) Joint Program, 2008
Title: Home-based Empowered Living for Parkinson’s disease Patients (HELP)
Responsibles: Prof. Giuseppina Campisi, Dr. Roberto Monastero
Amount of fund: .........................
Lenght: 3-year project
Abstract
Medical and social development have produced remarkable changes in the population distribution at
worldwide level, leading to a dramatical increase in the life expectancy of the human population. the higher
life expectancy is usually associated with the appearance of chronic conditions, such as Parkinson's disease
(PD), and associated comorbidites, which burdens and have a great impact in the quality of life of both the
persons suffering the illness and his/her relatives and caregivers. Besides, chronic conditions also derive in
higher costs for the care delivery process, becoming a problem for the whole society. In this sense, the
technological advances of the last decades can, and should, provide solutions to better manage the
treatment for the patients, reducing the social and economic impact of the chronic conditions. The Homebased Empowered Living for Parkinson’s disease Patients project will include a system able to administer
drug therapy in a controlled and either continuous or on-demand basis, to control disease progression and
to mitigate PD symptoms. Accordingly, it will give a comprehensive service to help Parkinson's patients to
better cope with their illness and reduce comorbidity.
Country: ITALY
Contact person: Alessandra Pedrocchi (Politecnico di Milano)
Date: 14th Jan 2010
I)
Strategic Issues
TOPIC 1
Neuro-rehabilitation and brain functional imaging: rehabilitation customization driven by best
exploitation of cerebral plasticity and health system sustainability
The spreading of neuro-motor pathologies has a great impact on the healthcare system European
Countries, being among the first causes of death (stroke is the third) but utmost being the first case of high
severity long-term disabilities. The recovery and the progress of neuromotor diseases is strongly affected
by the pathogenesis but also by the rehabilitation training, which has the purpose to best exploit the
cerebral plasticity governing neural motor control reorganization.
Beside the traditional therapist assisted training, different novel solutions has spread in the latest years
based on robotic systems, functional electrical stimulation (neuroprostheses) and hybrid systems where
robots and electrical stimulation are integrated.
Nonetheless, the international scientific community still lack of a complete well supported comparative
validation of these approaches and consequently a specific customization of the best training cocktail for
each patient is still not documented by scientific results and it is mainly devoted to clinician personal
convictions and experience.
Especially, we claim that in term of motor recovery and motor relearning, i.e. cerebral re-organization,
there is no clear scientific substrate to support the use of robotic devices and/or neuroprostheses aside or
in alternative to standard rehabilitation.
However, in the latest decade the possibility to investigate in-vivo cerebral activations associated to
movement execution has become accessible , thanks to fMRI, EEG and fNIRS,…
The challenge to be tackle in the next future is to integrate these two aspects aiming at studying the
impact of rehabilitation selections on the cerebral plasticity reorganization in neuromotor pathologies.
Beside the efficacy of those methods and their best exploitation, it also needs to be tested the
sustainability of these solutions for the long term economical, environmental and social impact of
biomedical technologies in the rehabilitation of neuromotor pathologies.
Nome
Alberto Pilotto
Contatti
Phone: +39 0882 410271
Fax:
+39 0882 410271
E-mail: [email protected]
Unità Operativa di Geriatria & Laboratorio di Gerontologia-Geriatria
Dipartimento di Scienze Mediche
IRCCS Casa Sollievo della Sofferenza
Viale Cappuccini, 1 – 71013 San Giovanni Rotondo (FG)
Istituto/Dipartimento
Proposta di ricerca
Sporadic Alzheimer’s disease (AD) is a progressive neurodegenerative disorder occurring predominantly in
older age. The prevalence of AD rise from 20% after 75 years to 30% after 85 years, and about 5% of
people aged 65 years or older have AD. With about 3 to 4 million people affected in the United States and
about 350,000 new cases per year, AD is the most frequent cause of dementia in U.S. and in Western
countries. Among the main cause leading to AD, the deficit of cholinergic system plays a major role.
Accordingly, one of the most common therapy for the symptomatic treatment of AD is the block of
acetylcholinesterase (AChE) by means of acetylcholinesterase inhibitors. The inhibition of AChE increases
the concentration of acetylcholine (ACh) in the synaptic cleft, thus restoring the physiological effects of
ACh within the central nervous system. Recent studies reported a significant benefits of
acetylcholinesterase inhibitors vs placebo on cognitive function, activities of daily living, and behavior.
These improvements, however, are not always detectable in clinical practice. Thus, it has been recently
suggested that the detection of improvements on cognitive function, activities of daily living, and behavior
may be obtained by the concomitant use of drugs acting on other pathogenetic AD mechanism. In
particular the European Medical Agency (EMEA) recommend the use of the AChE inhibitors donepezil and
rivastigmine in mild-to-moderate AD, and the addition of memantine in moderate-to-severe AD.
Memantine block the glutamate receptor, limiting the uncontrolled entrance of Ca2+ ion in the
postsynaptic neuron, thus delaying the Ca2+-excess neurodegeneration. Most studies reported that
interindividual differences in response to these drugs may be due to variability in drug metabolism related
to behavioral, clinical, and genetic factors, mainly hereditary polymorphisms of drug-metabolizing and
drug-transporting enzymes. Final objective of this proposal is to identify the genetic component
underlying the response/non response to the most common drugs currently used in the treatment of AD,
i.e. donepezil, rivastigmine and memantine, throughout the following specific objectives: 1) identification
of significant difference in the distribution of the genotypes of the CYP2D6 gene polymorphisms among
patients responders/non-responders to donepezil treatment; 2) identification of the specific DNA
alteration producing modifications in the CYP2D6 enzyme activity; 3) confirmation of the modification of
the enzyme activity by means of donepezil dosage in vivo; 4) identification of significant difference in the
distribution of the genotypes of the acetylcholinesterase gene polymorphisms among patients
responders/non-responders to rivastigmine treatment; 5) identification of the specific DNA alteration
producing modifications in the acetylcholinesterase-mediated hydrolysis of rivastigmine; 6) confirmation
of the modification of the enzyme activity by means of rivastigmine dosage in vivo; 7) identification of
significant difference in the distribution of the genotypes of the OCT2 gene polymorphisms among
patients responders/non-responders to memantine treatment; 8) identification of the specific DNA
alteration producing modifications in the OCT2 transporter of memantine; 9) confirmation of the
modification of the enzyme activity by means of memantine dosage in vivo. We expected that the
identification of functional polymorphisms in the CYP2D6, AChE and OCT2 genes may influence the clinical
efficacy of donepezil, rivastigmine and memantine in AD patients, and may be useful in identifying
subgroups of AD patients with different clinical response to treatment. Criteria and indicators to verify
results are: 1) the number of patients enrolled in the study in the first 6 months; 2) the number of
genotypes investigated at the end of the 1th year of the project; 3) the number of specific gene alteration
influencing the enzyme activity at the 2nd year of the project; 4) the quantitative data obtained from
plasma drug dosage at the end of the 2nd year of the project. This project is coherent with the ministerial
guidelines for the I.R.C.C.S. and well integrate the clinical practice in neurology.
Area di interesse identificata
Pharmacogenetics of drugs for Alzheimer’s Disease
Finanziamenti ricevuti
Titolo progetto
Ente finanziatore
Effect of weight loss on metabolic, functional, cognitive status and
biological markers of longevity in obese frail elderly
Ministero della Salute
Durata progetto
Two years
Abstract del progetto
Obesity, and in particular abdominal obesity causes serious metabolic and
medical complications and impairs quality of life. Moreover, in elderly
persons, obesity can lead to frailty by promoting chronic inflammation
and by exacerbating the decline in strength, endurance, balance and
mobility associated with aging and physical inactivity. Weight loss,
induced by calorie restriction and/or physical exercise, simultaneously
improves multiple metabolic risk factors for cardiovascular disease and
other medical abnormalities associated with obesity, and reduce
morbidity and mortality. However, the appropriate treatment for obesity
in elderly persons is controversial because of the potential harmful
effects of weight loss on bone and muscle mass. It has been reported that
weight loss can also modify biological pathways at the cellular level.
Insulin and of insulin-like growth factor-1 (IGF-1) have been involved in
cellular caloric equilibrium and other mechanisms, i.e. oxidative stress
and inflammation. Thus genetic polymorphisms in genes involved in these
metabolic pathways such as apolipoprotein E (APOE), sterol regulatory
element-binding protein cleavage-activating protein (SCAP), peroxisome
proliferators-activated receptor gamma-2 (PPR), -2B-adrenergic
receptor (ADRA2B), acyl-CoA synthetase 5 (ACSL5) and interleukin-6 (IL-6)
may influence the effect of weight loss on biological mechanisms and
possibly on functional and cognitive status in the frail elderly. It is possible
that weight loss in obese elderly persons can be beneficial by improving
metabolic, functional and cognitive status, or harmful by causing a
decrease in muscle and bone mass. However, the clinical and
physiological effects of weight loss in obese elderly subjects have never
been carefully evaluated. Thus, appropriate treatment for this rapidly
increasing segment of the elderly population remains controversial. The
purpose of this project is to advance our scientific knowledge by
determining the effects of weight loss on metabolic, functional and
cognitive status in obese elderly subjects. Moreover, we will evaluate the
role of weight loss on biological markers of metabolic health and
longevity, i.e. serum level of Insulin, insulin-like growth factor-1 (IGF-1),
transforming growth factor- (TGF-), tumor-necrosis factor- (TNF-),
interleukin-1 (IL-1) and C-Reactive Protein (CRP), and their relationships
with genetic polymorphisms in APOE, SCAP, PPR-, ADRA2B, ACSL5 and
Titolo progetto
IL-6 genes. As biomarker of longevity we will also evaluate the
accumulation of mitochondrial (mt) DNA mutation through the analysis of
the D310 mononucleotide repeat of mtDNA.
Smart Home for Elderly People (HOPE)
Ente finanziatore
Ministero della salute/Ministero dell’Università e della Ricerca Scientifica
Durata progetto
Two years
Abstract del progetto
Since Europe’s and worldwide population grows older, the need for care
is growing as well as people spend more money to get it. The solution
Smart Home for Elderly People (HOPE) promises to reduce the need for
carers, improve the life of older people and cut the cost of assistance. The
main objective of the “Hope” project is to produce an Integrated
Computer Technology (ICT) solution that will help the elderly people,
specifically those that suffer with the Alzheimer s disease (AD), achieve a
richer and more independent lifestyle. Dementia causes long and
oppressive suffering to patients and their relatives and imposes
enormous costs on society. About 25 million people suffered from
dementia in recent years. As a 4-fold increase of this number is expected
by 2050, dementia is one main health issue of the next decades. AD
covers 50-70% of all dementia cases, no cure exists, and effective and
reliable early diagnostic techniques are lacking. Early diagnosis and
progress monitoring of AD is a central part of treatment until future drugs
and prevention strategies become available. Elderly people with AD
spend most of their time at home. The “HOPE” solution consists of an
integrated, smart platform that will enable the elderly people with AD to
use innovative technology for a more independent life, easy access to
information, monitor their health, and serve as a source of inspiration for
users as well as for people working with assistive devices. Moreover, it
will enable them to perform by themselves activities they were not able
to do before and which are important for their daily personal life. The
main advantage of the proposed system is that it provides a basis for
integrating services for the elderly population while they are at home.
HOPE is a budgeted solution that will be installed at the elderly people’s
homes, and will provide services for (a) life-long, self organized,
appropriate educational environment and access to information, (b) care
management and health support, (c) self monitoring and decision making.
Optionally, the user will be able to activate software which automatically
establishes the necessary interactive, triple-play connection for receiving
tele-help and tele-assistance from specialized service providers, doctors
or other medical personnel. The proposed application will be intelligent
enough to offer safety in terms of controlling efficiently the home
environment, economy in terms of controlling and decreasing the need
for external help, and convenience in terms of adjusting the operation of
connected sub-systems to achieve the best possible user experience.
Nome
Vincenzo Mario Bruno GIORGINO
Contatti
TEL. (WK) (0039) (0)11 6706093 e-mail [email protected]
Cell. 3280380926
Istituto/Dipartimento
Department of Social Sciences - Faculty of Economics - University of
Torino
Proposta di ricerca
A transdisciplinary oriented proposal from a social sciences perspective
From the bedside to the bench and return
The major challenge in the field of senile dementia is represented by the existent gap between different
disciplinary approaches, namely between bio-medical and social/human sciences. The project aims to
answer to this gap within a transdisciplinary perspective, mainly based on the effort to integrate
experiential data coming from social sciences methods with biomedical ones. The former are considered a
possible source of knowledge that, under certain conditions, can be meshed with the latter.
The research strategy is based on the recruitment of around 50 patients with mild cognitive
impairment or symptoms of early Alzheimer’s disease and their caregivers, both formal and informal.
Selection criteria of participants within a grounded theory methodology will be based on the greater
similarity, i.e. the most similar personal/social characteristics as well as of the context.
The research design is a mixed method approach oriented to collect data from the experiential
level principally through methods such as journals, systematic observation, in-depth interviews and focus
groups with the caregivers, both formal and informal. The same could be said for Alzheimer’s patients as
far as possible and, in this regard, first-person observation is of the utmost importance as it allows a
knowledge that is unavailable with usual third person methods; the former can be assessed with adequate
criteria as recent literature is showing.
One major assumption is that the loss of cognitive functions could not mean the loss of the
awareness of this painful condition; moreover, if the patient is usually unable to express his/her feelings, it
does not mean that he/she does not have feelings at all. So far, we can develop adequate tools of
systematic observation to explore these more subtle processes.
The project may include a two month period of an experimental training for caregivers in which
they could learn to practice some contemplative methods in their everyday life. This is a goal of
translational relevance that can include the patients, depending by their specific condition, and calling for
a specific kind of exercises. Following this perspective, we can set up two groups of around 25 caregivers
each. To the first group is offered a two month contemplative practice program, while the second will
follow the current standard service provision.
If in the literature social ties are recognized of major importance both at health promotion and
prevention level than during the support for the sick, they do not exist in a vacuum: they call for personal
commitment, i.e. emotional, cognitive and economic resources. Social support and self healing are here
intended as nurtured by a disciplined learned skills of contemplative techniques: see for example the
literature about TM and mindfulness approaches which provide a cultural maintenance to our biological
self.
As a further development, a contribution by an economist could develop a specific research
strategy for a study about the costs of this kind of intervention compared with standard ones.
A valuable consequence of this approach could also be appreciated at cognitive level: a fresh
understanding of difficult situations could emerge in everyday life by the caregivers or in the professional
realm, suggesting new opportunities of managing the interactional lived situation and opening the way to
new possible treatments. The hypothesis is that contemplative methods could enhance not only the
coping function but also the general interactional process related to the management of this illness,
including a better interaction and understanding with/of the patient’s experience, currently shadowed in
clinical research. One can envisages also the consequences that this kind of approach could have at the
governance level, in which caregivers and, when possible, patients can be co-producers of the healing
process, as recent NHS policy programs show in the UK. A broader encompassing framework based on an
experiential-empirical perspective could enhance the scientific development and open to more effective
intervention.
Vincenzo Mario Bruno Giorgino
Area di interesse identificata
Experimental methods oriented to personal experience
Transdisciplinary research
Sociology of health, of health profession and informal care