- AIRC Associazione Italiana per la Ricerca sul Cancro
PROPOSAL FORM 2007
Via Corridoni, 7 - 20122 MILANO
tel.02/7797217-275 - fax 02/7797259
email: [email protected]
per informazioni tecniche tel.02/7797224
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ASSOCIAZIONE ITALIANA PER LA RICERCA SUL CANCRO
Grant Proposal 2007
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TITLE PAGE
Principal Investigator's full Name and Qualification
Professore Martinelli Giovanni - Professore associato
Proposal Title
Mechanisms responsible for sensitivity and resistance of Ph-positive cells to tyrosine kinase inhibitors Type of Grant
Area
IG
Targeted Therapy
Sub Area
Budget 2007 (euro): 50.000,00 €
Estimated budget 2007 - 2009 (euro): 130.000,00 €
Ente/Università
Università di Bologna Dipartimento/Istituto/altro
Istituto di Ematologia e Ocologia Medica "L. e A. Seràgnoli" - Indirizzo
Via Massarenti, 9 Città + ZIP Code
40138 BOLOGNA (BO) Telefono
0516363829 Fax
0516364037 E-mail
[email protected] Authorized Administrative Official
Prof. Pier Luigi Lollini- Centro Interdipartimentale Ricerca sul Cancro "Giorgio Prodi" Address
Via Massarenti, 9 City + ZIP Code
40138 BOLOGNA (BO) Phone
051307532 Fax
051349655 E-mail
[email protected] Proponent's signature
Martinelli Giovanni Date
15/12/2006
Authorized Administrative Official's signature
Prof. Pier Luigi Lollini- Centro Interdipartimentale Ricerca sul
Cancro "Giorgio Prodi" Date
15/12/2006
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EVALUATION FORM (selection 2002-2006)
Principal Investigator's Full Name
Professore Martinelli Giovanni - Professore associato
Total Papers and Reviews
75
Total IF
386,93
Average IF
5,2
Papers First/Last or Corresponding Author
41
Total IF
222,884
Average IF
5,4
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ABSTRACT
Principal Investigator's Full Name
Professore Martinelli Giovanni
Institution and City
Università di Bologna BOLOGNA
Proposal Title
Mechanisms responsible for sensitivity and resistance of Ph-positive cells to tyrosine kinase inhibitors
Area
Targeted Therapy
Sub Area
Abstract in the next page
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Imatinib mesylate is now the first-choice agent in the treatment of chronic myeloid leukemia (CML)
patients. Imatinib is a potent and selective inhibitor of the Bcr-Abl tyrosine kinase, whose
deregulated activity is the main pathogenetic mechanism of CML and Philadelphia (Ph)-positive
acute lymphoblastic leukemia (ALL). However, imatinib therapy is extremely expensive and is not
successful in all patients due to the emergence of resistance which ultimately leads to clinical
relapse and may contribute to disease progression. Basic and clinical research have to focus now on
the understanding and overcoming of imatinib resistance in order to offer each single CML/Ph+
ALL patient a real "tailored" therapy. Thus, the present research programme aims at the
optimization of the therapy of Ph-positive leukemias through a research strategy acting at different
levels and addressing all the main aspects related to drug-sensitivity and drug-resistance to imatinib,
namely through:
- the optimization of the molecular monitoring of patients treated with imatinib or novel inhibitors,
in order to provide clinicians with a comprehensive panel of biological indicators allowing to
evaluate the real efficacy of the treatment and to rationally and timely (re-)assess the therapeutic
strategy;
- the characterization of new mechanisms of resistance to imatinib which have been poorly or never
investigated so far;
- the identification of novel targets for a molecular therapy of those patients for whom inhibition of
Bcr-Abl turns out to be insufficient;
- the assessment in vitro in pre-clinical models (cell lines, primary patient cells and mouse models)
of novel inhibitors or novel inhibitory strategies targeting Bcr-Abl itself, key signal transduction
molecules downstream of Bcr-Abl, or proliferative and antiapoptotic processes in general;
- the assessment in vivo, in newly diagnosed patients or in patients already known to be resistant to
imatinib, of those second-generation inhibitors which are already in clinical phase;
- the identification and validation of novel biological factors which may serve as predictors of drugresponse and drug-resistance.
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PROPOSAL MAIN BODY
Background and rationale:
The Ph chromosome resulting from the translocation t(9;22) (q34;q11) represents the most common
cytogenetic abnormality detected in human leukemias. It is considered the hallmark of chronic
myeloid leukemia (CML), but can be detectable in 20-30% of adult acute lymphoblastic leukemia
(ALL) and in 4% of pediatric ALL, respectively. At the molecular level, the t(9;22) translocation
parallels the fusion of the proto-oncogene ABL with the BCR gene, resulting in the production of
the chimeric BCR/ABL gene, which leads to the synthesis of the fusion proteins (p210, p190)
which show a constitutive tyrosine kinase activity[1, 2]. Although the BCR/ABL hybrid gene is
characterized by a restricted number of molecular variants, the two main diseases, CML and ALL,
marked by this molecular rearrangement are different in terms of clinical and hematologic
characteristics. CML is a chronic myeloprolipherative disorder due to the massive expansion of
BCR/ABL+ myeloid cells, which, during the chronic phase, maintain the capacity to differentiate
normally. The progressive loss of normal differentiation capacity results in disease progression to
an acute leukemia or blast phase, which may show a myeloid (70%) or lymphoid phenotype. Ph+
and/or BCR/ABL+ ALL (almost all of the B-lineage) is, from the beginning, a disease with a very
aggressive behavior and poor prognosis. Turning off the aberrant tyrosine kinase activity of this
fusion protein means suppressing Ph+ cells. Therefore, Ph+ leukemic cells represent the ideal target
for tailored therapies based on tyrosine kinase inhibitors. Imatinib mesylate (IM) was developed as
the first molecularly targeted therapy that specifically inhibits the Bcr-Abl tyrosine kinase
activity[3-9]. IM interacts with the ATP-binding site of ABL only when the latter is in its inactive
conformation. In this way, IM is able to prevent the conformational transition to the active form,
which is responsible for binding and/or phosphorylation of signal transduction molecules. Due to its
excellent hematologic and cytogenetic responses, particularly in patients with chronic phase CML,
imatinib has moved towards first-line treatment for newly diagnosed CML. Despite the high rates of
complete cytogenetic response, resistance to the drug has been frequently reported[10-13].
Depending on the time of onset, two categories of resistance can be distinguished: if there is no
response after initial treatment, resistance is described as primary or intrinsic, in contrast,
secondary or extrinsic resistance is present if resistance develops after achieving an objective
response. The mainly mechanisms of resistance are the following:
Point mutations:
In the majority of cases, resistance is caused by reactivation of BCR-ABL tyrosine kinase activity
due to the emergence of specific point mutations within several critical regions of the Abl kinase
domain [14-20]. Such mutations impair IM binding either by affecting critical contact residues or
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by inducing a BCR-ABL conformation to which IM is unable to bind. More than 30 different point
mutations encoding for distinct single amino acid substitutions in the BCR-ABL kinase domain
have been identified in relapsed CML patients and Ph+ ALL. Different mutants seem to have
different degrees of resistance to imatinib: in vitro data indicate that while some mutations might be
overcome by dose escalation [21], other confer a highly resistant phenotype, thereby suggesting
withdrawal of imatinib in favour of alternative therapeutic strategies. Indeed, since resistance often
coincides with reactivation of the kinase activity within the leukemic clone, either Bcr-Abl itself or
Bcr-Abl-triggered downstream signalling pathways remain good targets for molecular therapy.
Several novel second-generation inhibitors [22] have been synthesized and are now being evaluated
in international phase I-II trials. They include novel Bcr-Abl inhibitors like nilotinib (AMN107)[23], dual Src/Abl inhibitors like dasatinib (BMS-354825)[23-25] and SKI-606 [26],
proteasome inhibitors like bortezomib (PS-341)[27]. Pre-clinical studies have already assessed IC50
values of several novel inhibitors against almost all mutant forms of Bcr-Abl, showing a precise
spectrum of sensitivity against some mutants and resistance against others, and have also
hypothesized novel inhibitor-specific mutants which are likely to emerge, though all these findings
have not been confirmed in patients. Only one specific mutant (i.e., the T315I) will remain highly
problematic for clinicians since (a) it is highly resistant to imatinib as well as to almost all novel
Abl or Src/Abl inhibitors, including dasatinib and nilotinib [23], the closest to FDA and EMEA
approval; (b) it seems to be associated with a highly aggressive disease phenotype and particularly
poor prognosis. Therefore, recent studies [28, 29] have shown that MK-0457 (VX-680), a smallmolecule aurora kinase inhibitor, has in vitro activity against the T315I-Bcr-Abl. Our purpose is to
confirm the efficacy of MK-0457 in vitro against the T315I-Bcr-Abl and furthermore to investigate
its efficacy, in vivo, in patients with CML or Ph+ ALL carrying the T315I-Bcr-Abl mutation.
BCR-ABL gene amplification
Resistance to IM can also be caused by over-expression of the Bcr-Abl protein due to gene
amplification of the BCR-ABL gene. This mechanism, observed in a proportionally small number
of imatinib-resistant patients, was initially described in the LAMA84R cell line with a 4.6-fold
increase in mRNA level.
Abnormal signal transduction pathways
Bcr-Abl exerts its oncogenic effects in CML cells essentially by stimulating cell proliferation,
inhibiting apoptosis and altering cell adhesion to bone marrow stroma. Signal transduction cascades
involved in these cellular processes and activated by Bcr-Abl include, among others:
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- Ras;
- mitogen-activated protein kinase (MAPK) and its downstream effectors MEK and Erk;
- phosphatidylinositol-3 kinase (PI3K) and its downstream effector Akt;
- phospho-tyrosine-phosphatase (PTP ).
PTPs proteins play critical roles in many cellular process as the gene expression, the regulation of
the cell growth, proliferation, differentiation, cell cycle, and cell movement by the counterbalance
of the effect on the cell growth of the protein tyrosine kinases (PTKs). Previous reports have
demonstrated that some phosphatases have an oncosuppressive role in Ph+ cells [30, 31]. This
suggest that PTP
plays a critical role in the pathogenesis of CML and that it could have a
oncosuppressor effect in vivo.
Deletions of sequences located on chromosome 9
Deletions of sequences located on chromosome 9 close to ABL gene have been recognized as a key
factor for progression of CML and for reduced responsiveness to Imatinib and other therapy. To
address this issue we will screen the Ph-positive CML genome through the array-CGH
(comparative genomic hybridization) provided by Nimblegen System. Comparative Genomic
Hybridization (CGH) measures DNA copy number differences between a reference genome and
your sample genome.
ABC transporters over-expression
Blood and tissue concentrations of most drugs are influenced by interindividual variations in genes
encoding drug metabolizing enzymes (DMEs) and drug transporters. Cytochrome P450 enzymes
(CYPs) are thought to have evolved as a protective adaptive response against environmental toxic
effects. Imatinib is metabolized mainly by CYP3A4 and CYP3A5 isoforms, and to a lesser extent
by CYP1A2, CYP2D6, CYP2C19. Some imatinib metabolites have been shown to be produced by
CYP1A1 and CYP1B1. Imatinib transport into cells has been shown to be mediated by hOCT1
(human Organic Cation Transporter, isoform 1), also known as SLC22A1 (Solute Carrier family 22,
member 1) [32]. The OCT family mediates electrogenic and sodium-independent translocation of
organic cations or weak bases, i.e., molecules with a transient or permanent positive net charge at
physiological Ph, in both directions across the plasma membrane. The family comprises three
members (hOCT1, hOCT2 and hOCT3 also known as EMT, Extraneural Monoamine Transporter)
differing in tissue distribution and substrate specificity.
Pre-imatinib hOCT1 expression levels have been demonstrated to be significantly lower in patients
who remain >65% Philadelphia-chromosome positive by cytogenetics during the first 10 months of
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imatinib treatment[33]. In contrast, imatinib efflux is thought to be mediated by ABC transporters
ABCG2 (also known as BCRP) and ABCB1 (MDR1; P-glycoprotein)[4, 34, 35]. The MDR-1 gene
is commonly over-expressed in blast cells of patients in the advanced phase CML [35, 36].
Genomic instability
BCR-ABL has been associated with genomic instability, which may have particular relevance
during disease progression from chronic phase to accelerated and blast phase CML. Recent findings
have proposed that most protein-enconding genes may be regulated by small RNAs that can
specifically control transcript turnover (siRNA) and/or protein translation (miRNA). In the latter
case miRNA can do such a job by blocking access or sliding of ribosomes to mRNAs, thus
impeding translation.
Alternative or aberrant spliced transcripts
Alternative splicing is the process whereby identical pre-mRNA molecules are spliced in different
ways, and this is important in normal development as a means of creating protein diversity in
complex organisms[37]. It is evident that alternative splicing plays a key role in biology: tissuespecific and developmental stage-specific alternative splicing contributes to significant protein
diversity; disease-related deregulation of splicing may be critical in pathogenesis and contribute to
disease diversity and complexity.
Pre-mRNA splicing is a sophisticated and ubiquitous nuclear process, which is a natural source of
cancer-causing errors in gene expression. Intronic splice site mutations of tumor suppressor genes
often cause exon-skipping events that truncate proteins just like classical nonsense mutations.
Spliced isoforms lacking critical N-terminal zinc-finger of the Ikaros transcription factor act as
dominant negatives by binding long isoforms through the C-terminal zinc-finger domain. Forced
expression of short isoforms (Ik4-Ik8) in murine or human hematopoietic progenitors arrest lineage
commitment and differentiation and play a role in the development of haematological malignancies,
such as ALL and crisis blastic CML[38, 39]. BCR-ABL1 kinase activity is also linked to the
expression of a truncated isoform of the adaptor protein SLP-65 and of the Bruton tyrosine kinase
(BTK)[40], which may contribute to the compromised pre-BCR signalling in ALL.
Src-family kinases
Another resistance mechanism could be the compensation of loss of BCR-ABL signalling by other
tyrosine kinase-mediated pathways. Src-family kinases such as Lyn are involved in BCR-ABLmediated leukemogenesis and have been found to be up regulated in cultured CML cells selected
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for IM resistance; increased Lyn expression was also found to correlate with clinical evidence of IM
resistance in some patients, which highlights the potential clinical relevance of this resistance
mechanism [20, 21]. The challenge for the future is to improve current clinical results with tyrosine
kinase inhibitors in Ph+ leukemias, developing strategies that can eradicate residual disease and
overcome or prevent resistance.
Description of the project:
The present research program can be subdivided in four tasks:
1) Collection and storage of biological material for molecular and cellular studies.
The Research Unit-Martinelli belongs to the Institute of Hematology and Medical Oncology
"Seràgnoli" which is the leader of the GIMEMA Working Party on CML, and as such, is
coordinating or participating in several national and international clinical trials with imatinib and
novel inhibitors. Upon written informed consent of the patients enrolled in these trials, the Research
Unit will create a precious bank of:
a) biological samples, regularly collected after written informed consent of the patient, processed
and stored at baseline and at regular time points during treatment; material includes mononuclear
cells and CD34+ cells, protein lysates, RNA, DNA.
b) clinical data, accurately recorded in an electronic format at baseline and at regular timepoints
during treatment and available for correlations between biological findings and clinical outcome.
Mechanisms of CML and Ph+ ALL resistance will be also investigated on all resistant patients
treated with IM or other tyrosine kinase inhibitor not in the framework of clinical trials upon written
informed consent.
The biological material collected with the cell lines and with the mouse models will create a solid
basis for all the planned studies.
2) Optimization of molecular monitoring of patients treated with imatinib and with novel
inhibitors, in order to evaluate the actual efficacy of the treatment and to rationally and
timely (re-)assess the therapeutic strategy.
a) Screening for ABL KD mutations
The molecular biology laboratory of the present research unit has already developed a rapid and
reliable screening method for Abl KD mutations based on a novel high-throughput denaturing-high
performance liquid chromatography (D-HPLC) device with UV detector (D-HPLC 3500HT;
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Transgenomic), which is now available to all the Institutions of the GIMEMA Working Party on
CML. D-HPLC is a reversed-phase, ion-pairing HPLC, specifically developed for the detection of
DNA sequence variations such as point mutations, small insertions and deletions. Under conditions
of partial heat denaturation, heteroduplexes that form in PCR samples with internal sequence
variations display reduced column retention with respect to their homoduplex counterparts.
Therefore, the elution profiles for such samples are distinct from those with a homozygous
sequence, making the identification of samples harbouring mutations a rapid (~3 min) and
straightforward procedure. Samples scored positive by D-HPLC are then sequenced in order to
characterize the exact nucleotide substitution. The present unit will implement the protocol of
mutation screening by setting up and standardizing a mutation detection method based on a DHPLC equipped with a novel fluorescence detector, allowing to increase the sensitivity of detection
of at least tenfold, from 5% to 0.5-0.1%.
b) A T315I-specific assay
Given the importance of an early detection of T315I-positive mutant clones which render imatinib,
dasatinib and nilotinib treatment ineffective and have been associated with a highly aggressive
disease phenotype, we will also develop a sensitive and straightforward diagnostic method specific
for T315I allowing for a rapid and accurate detection of this mutation. To this purpose, we will setup three different approaches, which will be tested and compared in terms of sensitivity, specificity
and reliability:
- a conventional allele-specific oligonucleotide (ASO)-PCR approach, with forward primers
designed on BCR sequences (exon 1 for p190, exon 13 for p210) and two sequence-specific reverse
primers, one for the wild-type and one for the mutant ABL sequence, followed by gelelectrophoresis analysis of amplification products;
- an allelic discrimination assay based on two fluorescent probes specifically designed to anneal to
the wild-type and mutated sequence, during a multiplex Q-PCR reaction on the ABI-PRISM 7900
(Applied Biosystems);
- an assay based on PCR amplification of a Bcr-Abl fragment encompassing codon 315 followed by
denaturation at 96°C, gradual reannealing at room temperature, digestion with Surveyor Nuclease
(Transgenomic) which cuts heteroduplexes if and where a mismatch exists, and separation of cut or
uncut fragments by gel-electrophoresis or by D-HPLC elution.
c) Molecular monitoring of the BCR-ABL transcript levels and of the eventual BCR-ABL
over expression , so as to determine whether resistance may be caused also by gene-dosage effects.
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BCR-ABL transcript levels will be monitored by real-time Taqman RT-PCR; we will determinate if
there is an amplification of BCR-ABL by fluorescence in situ hybridization (FISH).
d) Analysis of the karyotype of Ph+ cells , so as to identify the additional cytogenetic
abnormalities responsible for resistance. These additional cytogenetic abnormalities associated with
resistance will be investigated both by conventional and by molecular (FISH, fluorescence in situ
hybridization) cytogenetic techniques.
3) Characterization of mechanisms of resistance to imatinib and novel TK inhibitors.
a) Abl KD mutations in patients resistant to imatinib treated with dasatinib, nilotinib, SKI606 and MK-0457
In order to assess the degree of sensitivity of various Abl KD mutations in vivo in patients treated
with novel tyrosine kinase inhibitors, as well as the likelihood of emergence of novel, inhibitorspecific mutant forms, we will regularly perform mutation monitoring of patients resistant to or
intolerant of imatinib treated at the "Seràgnoli" Institute with dasatinib, nilotinib, SKI-606 and MK0457. Patients will be analyzed at baseline and every month thereafter, in order to follow the
kinetics of disappearance of pre-existing mutated clones or the kinetics of selection of novel ones.
Mutations observed in vivo will be assessed for their biological-structural effects on Bcr-Abl, on its
tyrosine kinase activity, on the interaction with downstream signaling molecules and with inhibitors
themselves again using specific computer-assisted molecular simulation techniques. We will also
develop softwares allowing to rapidly and reliably predict the effects of any reported or unreported
mutation on the novel tyrosine kinase inhibitors currently under development or still in a preclinical phase.
b) Analysis of signal transduction pathways regulated by PTP
This will be accomplished following two complementary approaches:
- Identification of substrates
We plan to analyze selected p210 BCR/ABL substrates in order to link the observed effect to a
molecular mechanism. Whether the dephosphorylation of p210 BCR/ABL or other potential
substrates is directly or indirectly mediated by PTP will be investigated by the "substrate trapping"
approach, as binding of substrates to active PTPs is usually too weak to allow the detection of their
interaction using standard co-immunoprecipitation protocols. This technique is based on the
observation that PTPs are defined by the presence of a signature sequence motif,
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[I/V]HCXXGXXR[S/T]. It is possible to obtain PTPs that maintain a high affinity for substrate but
do not effectively catalyze dephosphorylation, thus converting an extremely active enzyme into a
substrate trap. We will obtain the mutant cDNA (D198A-PTP ), now sequencing for the quality
control, and we plan to use it for transfection experiments.
Methods:
Cells transfected with PTP cDNA WT or D198A-PTP will be lysed with cell disrupting medium
capable to separate specific cellular fractions (FOCUS Phosphorich and Fraction FOCUS kits from
Geno-Tech, St. Louis, MO) and immunoprecipitation. Proteins linked to TAT-mut-ICD will be
analyzed by SDS-PAGE/ Western Blotting, 2-DGE or directly subjected to 2DC-MS/MS analysis.
We will analyze immunoprecipitated protein. D198A-PTP will be also immunoprecipitated and
analysed by western blotting with specific antibodies in order to identified proteins interacting with
PTP .
- Global analysis of gene expression
Microarray analysis is a powerful tool that enables to monitor the expression profile of thousands of
genes in a single assay. In this way it is possible to affiliate a specific gene expression profile to a
certain cell line or patient's sample and so potentially to identify the genes associated to a particular
phenotype or disease. We plan to study the effect of PTP expression in K562 stable transfectants in
the presence or absence of Imatinib. A total of 12 hybridization reactions are planned (triplicate
assays for each of the four experimental conditions planned).
Methods:
For isolation of total cellular RNA, 5x10(6) cells, cultured in the absence or presence of the
inducing agents indicated, will be harvested and RNA will be prepared using standard methods.
The cDNA hybridization and the collection and analysis of the data will be performed by CRIB,
Padova, Italy (visit http://microcribi.cribi.unipd for details). Validation of identified targets will be
performed by QPCR followed, whenever possible, by immunodetection.
c) Expression and genotyping of single nucleotide polymorphisms (SNPs) in imatinib
transporters and metabolizing enzymes as predictors of outcome of CML patients
Since interindividual variation(s) in these genes encoding may influence - and may therefore allow
to predict - the outcome of CML patients treated with tyrosine kinase inhibitor, our aim is to
correlate SNPs in key genes to imatinib efficacy and to identify one or more SNPs with predictive
value allowing to optimize the therapeutic use of imatinib in CML patients. To this purpose, we will
assess a set of SNPs as predictors of imatinib efficacy. To this purpose, we will focus on key genes
encoding for: DMEs (CYP3A4, CYP3A5, CYP1A1, CYP1B1, CYP2D6, CYP2C19), transporters
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(hOCT1, ABCB1, ABCG2). Genomic DNA will be isolated from 1-2 cc of peripheral blood or
bone marrow samples by conventional methods. Polymorphisms will be analyzed by polymerase
chain reaction (PCR) combined with restriction fragment length polymorphisms (RFLP) assay.
d) Study on promoter hypomethylation of the LINE-1 retrotransposable elements which
activates sense/antisense transcription and marks the progression of CML
The main aim of this part of the proposal is to identify miRNAs that can specifically interfere with
BCR and ABL. This will be achieved though bioinformatic tools that scan the human genome for
short DNA sequences with complete or partial homology to the BCR or ABL gene. One of such
programs is provided freely by the Computational Biology Center of Memorial Sloan-Kettering
Cancer Center. Putative miRNAs that are predicted to affect BCR and ABL regulation will be
cloned, expressed in human cells and tested for the ability to negatively affect BCR and ABL
translation. Those that will be found positive to the assay will also be used either alone or in
combination to affect translation of the chimeric BCR-ABL transcript.
e) Gene expression profiling
In order to identify molecular pathways that may be down/up-regulated by exposure to novel
tyrosine kinase inhibitors, we will perform a DNA microarray analysis on cell lines and on CD34+
cells from imatinib-resistant CML patients before and after treatment with other inhibitors, such as
MK-0457. This approach will allow us to elucidate the mechanisms responsible of the potential
efficacy of MK-0457 in CML cells. We will also use this gene expression profiling strategy to
identify a genomic profile that may be associated with sensitivity or resistance to MK-0457.
6
As the median CD34+ cells number in CML patients at diagnosis is usually between 0,5 and 1x10
(0,1% of mononucleated cells), we should expect to obtain a wide range of RNA quantity, whose
minimum will be presumably around 1,5 g (diluted in 30
l). Therefore, a double RNA
amplification should be planned, in order to obtain a sufficient cRNA quantity to efficiently
hybridize the Affymetrix chip.
Methods will be as follows:
a. CD34+ cell fraction separation: Mononuclear cells from PBL or BM samples will be isolated
by density gradient centrifugation (Ficoll separation) , and CD34+ cell fractions will be selected
by binding to immunomagnetic beads (AutoMACS; Miltenyi Biotech).
b. RNA extraction: Total RNA will be extracted from CD34+ cells using the Qiagen RNeasy
Mini or Micro Kit (Qiagen); .RNA quantity will be assessed by Nanodrop analysis (Nanodrop
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Technologies), while the Bioanalyzer (Agilent) will be used to assess the quality of the RNA
before starting with the synthesis of cRNA.
c. Synthesis of Biotin-Labeled cRNA: Biotin-labeled target synthesis reactions will be performed
using standard protocols supplied by the manufacturer (Affymetrix). Briefly, 100ng of the RNA
will be converted into double-stranded cDNA by reverse transcription using the cDNA synthesis
kit, following the protocol supplied by the manufacturer, with a T7-(dT)24 primer (Affymetrix).
After the second-strand synthesis, cRNA will be generated from the purified cDNA sample
(Gene Chip Sample Cleanup Module; Affymetrix) by an in vitro transcription reaction
(MEGAscript® T7 Kit, Ambion). The cRNA will be then purified (Gene Chip Sample Cleanup
Module; Affymetrix) and a second cycle of
sample cleanup
retrotransciption, second-strand synthesis and
will be performed, followed by a final Biotin-Labeled cRNA synthesis
(GeneChip IVT Labeling Kit, Affymetrix). The labelled cRNA will be purified using the
Affymetrix spin columns and the concentration of biotin-labeled cRNA will be determined by
Nanodrop, while the quality check will be performed by means of the Bioanalyzer.
d. Hybridization: 5 g of each biotinylated cRNA preparation will be fragmented and put in the
hybridization cocktail. Samples will be hybridized to Affymetrix HG133 2.0 Plus Gene Chip
Arrays for 16 hours. Gene chips will be than washed and stained following the instruments
standard Eukaryotic GE WS2v4 protocol and using antibody-mediated signal amplification.
e. Data analysis: The images from the scanned chips will be processed by means of Affymetrix
Microarray Analysis Suite 5.0 (MAS 5.0). The amount of a transcript mRNA will be determined
with the MAS 5.0 absolute analysis algorithm, as well as the presence or the absence of a
transcript. The identification of differentially expressed genes and the patients clustering will be
performed with GeneSpring 7.3 software (Silicon Genetics, Redwood City, CA). The
identification of biologic processes, molecular functions and cellular components of genes will
be assessed by EASE and Ingenuity® softwares.
f) Analysis of alternative or aberrant spliced transcripts in Ph+ ALL and CML patients
Studies in human Ph+ positive leukemias have gained newinsight into the essential role of truncated
protein isoforms in the pathogenesis and disease progression of CMl and BCR-ABL positive ALL,
but many challenges remain. Our aim is to analyze if leukemia-specific alternative or aberrant
splicing in BCR-ABL or other genes involved in BCR-ABL signaling such as Ikaros, BTK, SPL65
and other could be associated with resistance to imatinib or new tyrosine kinase inhibitors (TKI)
such as dasatinib or nilotinib. We will perform a GeneChip® Exon Array System, which are the
first experimental tools available to survey both gene expression and alternative splicing patterns on
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the whole-genome scale on a single array. This approach will deepen the understanding of biology
for many discovery-focused applications including: analysis of molecular mechanisms regulated by
alternative splicing; discovery of new splice variants of drug targets and mapping of their tissuespecific expression; improved understanding of the downstream effects of genetic variations that
may result in phenotypic changes in gene expression or alternative splicing patterns. We will
validate our results performing reverse transcription-polymerase chain reaction (RT-PCR) and
nucleotide sequencing.
4) In vivo testing, in patients with newly diagnosed CML o in patients resistant to imatinib at
standard dose, of imatinib at high doses or second-generation inhibitors already in clinical
phase
The feasibility and efficacy of therapeutic strategies alternative to the administration of imatinib at
standard dose will be tested in vivo in patients with CML or Ph-positive ALL. As the coordinator of
the GIMEMA Working Party on CML, the "Seràgnoli" Institute is leading and will lead two clinical
trials with imatinib at high dose, namely:
a) CML/021/STI571: newly diagnosed CP patients, high Sokal risk, treated with imatinib 800 mg/d;
b) CML/022/STI571: newly diagnosed CP patients, intermediate Sokal risk, randomized to receive
either imatinib 400 mg/d or imatinib 800 mg/d.
Moreover, the "Seràgnoli" Institute is actively participating and will participate in several
international phase I-II clinical trials with novel tyrosine kinase inhibitors, namely:
c) CA180005, CA180006, CA180013, CA180015, CA180035, phase II trials with dasatinib in
CML and Ph+ ALL patients resistant to or intolerant of imatinib;
d) CAMN107A2101, a phase II trial with nilotinib in CML and Ph+ ALL patients resistant to or
intolerant of imatinib;
e) 3160A4-200-WW, a phase I-II trial with SKI-606 in CML and Ph+ ALL patients resistant to or
intolerant of imatinib.
In addition to these ongoing trials, additional trials are planned to be activated by the GIMEMA
Working Party on CML itself by the end of 2006, namely:
f) a phase II trial with nilotinib administered first-line in newly diagnosed CP patients;
g) a phase I-II trial with bortezomib in CML patients resistant to imatinib;
h) a phase II trial with homoarringtonine in CML patients resistant to imatinib with evidence of the
T315I mutation.
Codice Riferimento: 4121
Page 16 of 44
Based on the results of some of the studies described in the sections above, we hope to identify
several biological predictive factors, potentially useful to predict the likelihood of response or
resistance to imatinib therapy.
The whole project will be performed in three years according to the phases and the timing specified
in the scheme below.
References:
1.
2.
Bartram C.R. dKA, H.A., et al., Translocation of c-abl oncogene correlates with the
presence of a Philadelphia cromosome in chronic myelocytic leukaemia. Nature, 1983. 306:
p. 277-80.
Lugo TG, P.A., Muller AT, et al., Tyrosine kinase activity and transformation potency of
bcr-abl oncogene products. Science, 1990. 247: p. 1079-1082.
Codice Riferimento: 4121
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responses in patients with accelerated phase chronic myeloid leukemia: results of a phase 2
study. Blood, 2002. 99: p. 1928-1937.
Crossman LC, D.B., Deininger MW, Pirmohamed M, Wang L, Clark RE., hOCT 1 and
resistance to imatinib. Blood, 2005. 106: p. 1133-1134.
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mesylate in chronic myelogenous leukemia. N Engl J Med, 2002. 346: p. 645-652.
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chronic myeloid leukemia. Haematologica, 2003. 88: p. 256-259.
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early chronic-phase chronic myeloid leukemia. Blood, 2004. 104: p. 4245-4251.
Rosti G, M.G., et al. , Molecular response to imatinib in late chronic-phase chronic myeloid
leukemia. Blood, 2004. 103: p. 2284-2290.
O'Brien SG, G.F., Larson RA, Gathmann I, Baccarani M, et al. , Imatinib compared with
interferon and low-dose cytarabine for newly diagnosed chronic-phase chronic myeloid
leukemia. . N Engl J Med, 2003. 348: p. 994-1000.
Kantarjian HM, C.J., O'Brien S, Luthra R, Giles F, Verstovsek S, Faderl S, Thomas D,
Garcia-Manero G, Rios MB, Shan J, Jones D, Talpaz M., Long-term survival benefit and
improved complete cytogenetic and molecular response rates with imatinib mesylate in
Philadelphia chromosome-positive chronic-phase chronic myeloid leukemia after failure of
interferon-alpha. Blood, 2004. 104: p. 1979-1988
Iacobucci I, R.G., Amabile M, Poerio A, Soverini S, Cilloni D, Testoni N, Abruzzese E,
Montefusco E, Ottaviani E, Iuliano F, Russo D, Gobbi M, Alimena G, Martino B, Terragna
C, Pane F, Saglio G, Baccarani M, Martinelli G., Comparison between patients with
Philadelphia-positive chronic phase chronic myeloid leukemia who obtained a complete
cytogenetic response within 1 year of imatinib therapy and those who achieved such a
response after 12 months of treatment. JOURNAL OF CLINICAL ONCOLOGY, 2006. 24:
p. 454 - 459.
Martinelli, G., et al., Prediction of response to imatinib by prospective quantitation of BCRABL transcript in late chronic phase chronic myeloid leukemia patients. Ann Oncol, 2006.
17(3): p. 495-502.
Gorre ME, M.M., Ellwood K, Hsu N, Paquette R, Rao PN, Sawyers CL., Clinical resistance
to STI-571 cancer therapy caused by BCR-ABL gene mutation or amplification. Science,
2001. 293: p. 876-880.
Hochhaus A, K.S., Corbin AS, La Rosee P, Muller MC, Lahaye T, Hanfstein B, Schoch C,
Cross NC, Berger U, Gschaidmeier and D.B. H, Hehlmann R., Molecular and chromosomal
mechanisms of resistance to imatinib (STI571) therapy. Leukemia, 2002. 2002: p. 21902196.
Roche-Lestienne C, S.-C.V., Grardel-Duflos N, Lai JL, Philippe N, Facon T, Fenaux P,
Preudhomme C. , Several types of mutations of the Abl gene can be found in chronic
myeloid leukemia patients resistant to STI571, and they can pre-exist to the onset of
treatment. Blood, 2002. 100: p. 1014-1018.
Shah NP, N.J., Nagar B, Gorre ME, Paquette RL, Kuriyan J, Sawyers CL., Multiple BCRABL kinase domain mutations confer polyclonal resistance to the tyrosine kinase inhibitor
imatinib (STI571) in chronic phase and blast crisis chronic myeloid leukemia. Cancer Cell,
2002. 2: p. 117-125.
Branford S, R.Z., Walsh S, Parkinson I, Grigg A, Szer J, Taylor K, Herrmann R, Seymour
JF, Arthur C, Joske D, Lynch K, Hughes T. , Detection of BCR-ABL mutations in patients
Codice Riferimento: 4121
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with CML treated with imatinib is virtually always accompanied by clinical resistance, and
mutations in the ATP phosphate-binding loop (P-loop) are associated with a poor
prognosis. Blood, 2003. 102: p. 276-283.
Soverini S, M.G., Amabile M, Poerio A, Bianchini M, Rosti G, Pane F, Saglio G, Baccarani
M. , Denaturing-HPLC-based assay for detection of ABL mutations in chronic myeloid
leukemia patients resistant to Imatinib. Clin Chem., 2004. 50: p. 1205-1213.
Soverini S, M.G., Rosti G, Bassi S, Amabile M, Poerio A, Giannini B, Trabacchi E,
Castagnetti F, Testoni N, Luatti S, de Vivo A, Cilloni D, Izzo B, Fava M, Alberti D, Pane F,
Saglio G, Baccarani M., Abl mutations in late-chronic phase chronic myeloid leukemia
patients with upfront cytogenetic resistance to imatinib are associated with a greater
likelihood of progression to blast crisis and shorter survival. J Clin Oncol. 23: p. 4100 4109.
Corbin AS, L.R.P., Stoffregen EP, Druker BJ, Deininger MW. , Several Bcr-Abl kinase
domain mutants associated with imatinib mesylate resistance remain sensitive to imatinib.
Blood, 2003. 101: p. 4611-4614.
Martinelli, G., et al., Dual tyrosine kinase inhibitors in chronic myeloid leukemia.
Leukemia, 2005. 19(11): p. 1872-9.
O'Hare T, W.D., Stoffregen EP,et al., In vitro Activity of Bcr-Abl Inhibitors AMN107 and
BMS-354825 against Clinically Relevant Imatinib-Resistant Abl Kinase Domain Mutant.
Cancer Res, 2005. 65: p. 4500-4505.
Shah NP, T.C., Lee FY, Chen P, Norris D, Sawyers CL. , Overriding imatinib resistance
with a novel ABL kinase inhibitor. Science, 2004. 305: p. 399-401.
Sawyers CL, S.N., Kantarjian HM, et al. Proceedings of the 2005 ASCO Annual Meeting. ,
A Phase I Study of BMS-354825 in Patients with Imatinib-Resistant and Intolerant
Accelerated and Blast Phase Chronic Myeloid Leukemia (CML): Results from CA180002.
2005.
Golas JM, A.K., Etienne C, et al., SKI-606, a 4-anilino-3-quinolinecarbonitrile dual
inhibitor of Src and Abl kinases, is a potent antiproliferative agent against chronic
myelogenous leukemia cells in culture and causes regression of K562 xenografts in nude
mice. Cancer Res, 2003. 63: p. 375-81.
Gatto S, S.B., Pham L, et al. , The proteasome inhibitor PS-341 inhibits growth and induces
apoptosis in Bcr/Abl-positive cell lines sensitive and resistant to imatinib mesylate.
Haematologica, 2003. 88: p. 853-863.
Carter TA, W.L., Shah NP, Velasco AM, Fabian MA, Treiber DK, et al., Inhibition of drugresistant mutants of ABL, KIT, and EGF receptor kinases. Proc Natl Acad Sci U S A, 2005.
102: p. 11011-6.
Young MA, S.N., Chao LH, Seeliger M, Milanov ZV, Biggs WH 3rd, et al., Structure of the
kinase domain of an imatinib-resistant Abl mutant in complex with the Aurora kinase
inhibitor VX-680. Cancer Res, 2006. 66: p. 1007-14.
Chien W, T.N., Williamson EA, Shih LY, Krug U, Kettenbach A, Fermin AC, Roifman
CM, Koeffler HP, Characterization of a myeloid tyrosine phosphatase, Lyp, and its role in
the Bcr-Abl signal transduction pathway. J Biol Chem, 2003. 278: p. 27413-20.
Lim YM, W.S., Lau G, Witte ON, Colicelli J, BCR/ABL inhibition by an escort/phosphatase
fusion protein. Proc Natl Acad Sci U S A, 2000. 97: p. 12233-8.
Marull M, R.B., Fragmentation study of imatinib and characterization of new imatinib
metabolites by liquid chromatography-triple-quadrupole and linear ion trap mass
spectrometers. J Mass Spectrom, 2006. 41: p. 390-404
Thomas J, W.L., Clark RE, Pirmohamed M., Active transport of imatinib into and out of
cells: implications for drug resistance. Blood, 2004. 104: p. 3739-3745.
Ozvegy-Laczka C, H.T., Varady G, et al., High-affinity interaction of tyrosine kinase
inhibitors with the ABCG2 multidrug transporter. Mol Pharmacol, 2004. 65: p. 1485-1495.
Codice Riferimento: 4121
Page 19 of 44
35.
36.
37.
38.
39.
40.
Burger H, v.T.H., Brok M, et al., Chronic imatinib mesylate exposure leads to reduced
intracellular drug accumulation by induction of the ABCG2 (BCRP) and ABCB1 (MDR1)
drug transport pumps. Cancer Biol Ther, 2005. 4: p. 747-752.
Illmer T, S.M., Platzbecker U, et al., P-glycoprotein-mediated drug efflux is a resistance
mechanism of chronic myelogenous leukemia cells to treatment with imatinib mesylate.
Leukemia, 2004. 18: p. 401-408.
Kalnina Z, Z.P., Silina K, Line A., Alterations of pre-mRNA splicing in cancer. Genes
Chromosomes Cancer, 2005. 42: p. 342-57.
Nera, K.P., et al., Ikaros has a crucial role in regulation of B cell receptor signaling. Eur J
Immunol, 2006. 36(3): p. 516-25.
Klein, F., et al., BCR-ABL1 induces aberrant splicing of IKAROS and lineage infidelity in
pre-B lymphoblastic leukemia cells. Oncogene, 2006. 25(7): p. 1118-24.
Kersseboom, R., et al., Bruton's tyrosine kinase and SLP-65 regulate pre-B cell
differentiation and the induction of Ig light chain gene rearrangement. J Immunol, 2006.
176(8): p. 4543-52.
Codice Riferimento: 4121
Page 20 of 44
PERSONNEL INVOLVED IN THE RESEARCH
Name
Date of birth
Role on project
Fellowship
Required
Effort on
project
Man/year
effort
Professore Giovanni Martinelli
30/05/1960
PI
'RWWRUHVVD0DULOLQD$PDELOH
,QYHVWLJDWRU
'RWWRUHVVD6LPRQD6RYHULQL
5HVHDUFKHU
'RWWRUHVVD,ODULD,DFREXFFL
3K'6WXGHQW
'RWWRUHVVD7L]LDQD*UDIRQH
5HVHDUFKHU
'RWWRUH)DXVWR&DVWDJQHWWL
5HVHDUFKHU
'RWWRUHVVD6WHIDQLD3DROLQL
5HVHDUFKHU
'RWWRUHVVD6DEULQD&RODURVVL
5HVHDUFKHU
'RWWRUHVVD$OHVVDQGUD*QDQL
5HVHDUFKHU
'RWWRUHVVD)UDQFHVFD3DODQGUL
5HVHDUFKHU
Total Man/Year on project
Codice Riferimento: 4121
Page 21 of 44
DESCRIPTION OF THE WORK FOR EVERY UNIT OF PERSONNEL.
DESCRIPTION OF THE WORK FOR EVERY UNIT OF PERSONNEL
Dr. Giovanni Martinelli is the coordinator.
His role is to design, coordinate, promote the project. It’s mainly to participate to the optimization
of molecular monitoring of patients treated with imatinib and with novel tyrosine kinase inhibitors
(Task 2) and to the identification of new mechanisms of resistance (Task 3). As the coordinator he
is involved in the design of clinical studies in CML and Ph-positive ALL to test the feasibility and
efficacy of therapeutic strategies alternative to the administration of imatinib at standard dose.
Dr.ssa Marilina Amabile
Her role in the project is to collect and storage biological material for molecular and cellular
studies, to monitor the BCR-ABL transcript level by RQ-PCR (TaqMan methodology) and to
analyze the karyotype of Ph positive cells (Tasks 1 and 2) investigate the intracellular signal
transduction pathways activated by BCR/ABL in Ph-positive leukemias, to participate to the
identification of new diagnostic and prognostic molecular markers, and discovery of new and
molecular targets in Ph+ for future therapeutic intervention.
Dr.ssa Simona Soverini
She is involved in developing methods to implement the protocol of mutation screening and in
developing a sensitive and straightforward diagnostic method specific for T315I allowing for a
rapid and accurate detection of this mutation (Task 2). Her role is also to analyze Abl KD mutations
in patients resistant to imatinib treated with novel tyrosine kinase inhibitors and to assess a set of
SNPs in key genes as predictors of imatinib efficacy (Task 3).
Dr.ssa Ilaria Iacobucci
Her role is to collect samples from CML and Ph-positive ALL patients (Task 1) and to analyze
alternative or aberrant spliced transcripts of molecules involved in B-signaling (Task 3) to evaluate
whether these alterations could be associated with resistance to imatinib or new tyrosine kinase
inhibitors.
Dr. Fausto Castagnetti
Registration of clinical data and collection of sample from CML and Ph-positive ALL patients
(Task 4).
Codice Riferimento: 4121
Page 22 of 44
Dr.ssa Tiziana Grafone
In vitro studies with inhibitors. to match the molecular and phenotypic findings with the clinical
course of the disease aiming to possibly define specific risk factors associated with resistance, poor
disease free survival, and poor overall survival. Analysis of ignal transduction pathways regulated
by PTPγ.
Dr.ssa Stefania Paolini
Registration of clinical data and collection of sample from CML and Ph-positive ALL patients
(Task 4).
Dr.ssa Sabrina Colarossi
Her role in the project is to identify miRNAs that can specifically interfere with BCR and ABL
(Task 3).
Dr.ssa Alessandra Gnani
She will be involved in Task 3 and in particular in the section which is dedicated to gene expression
analysis on CML and Ph-positive ALL samples either exposed or not to TK inhibitors in vitro and
in vivo studies.
Dr.ssa Francesca Palandri
Her role in the project is to collect and storage biological material for molecular and cellular studies
(Task 1). She is also involved in the registration of clinical from CML and Ph-positive ALL patients
(Task 4).
Codice Riferimento: 4121
Page 23 of 44
BUDGET FORM
2007
Research costs
2008
2009
Total
50.000,00 €
40.000,00 €
40.000,00 €
130.000,00 €
Fellowships
0,00 €
0,00 €
0,00 €
0,00 €
Indirect costs
0,00 €
0,00 €
0,00 €
0,00 €
50.000,00 €
40.000,00 €
40.000,00 €
130.000,00 €
0,00 €
0,00 €
0,00 €
0,00 €
50.000,00 €
40.000,00 €
40.000,00 €
130.000,00 €
Subtotal
Overhead
Total
Justificatons
RESEARCH COSTS
- Informatic systems;
- oligonucleotides sintesis, TaqMan primers and probes, disposable materials, RNA extraction kit, retrotranscription kit, amplification
kit, purification and sequencing kit;
- data handling and analysis;
- missions for coordination and cooperation about grants and projects;
- scientific publications, abstracts, presentations, slides, divulgation of results, websites;
- organization of meetings; registration fees to attend national and international meetings and present the results of the research.
FELLOWSHIPS COSTS
Not requested.
INDIRECT COSTS
Not requested.
OVERHEAD COSTS
Not requested.
Codice Riferimento: 4121
Page 24 of 44
BIOGRAPHICAL SKETCH
BIOGRAPHICAL SKETCH
Name
Professore Martinelli Giovanni
Position
Professore associato
Date of birth
30/05/1960
Education and Training (include degrees and post-doctoral training)
Institution and Location
Degree
Year (from/to)
Field of Study
University of Verona - Verona
Graduated
1980/1985
Medicine
University of Verona - Verona
Specialization
1985/1988
Hematology
University of Verona - Verona
Specialization
1988/1992
Medical Genetics
RESEARCH AND PROFESSIONAL EXPERIENCE
Year (from/to)
Institution
Position
City
Country
1984/1988
University of
Verona-Institute of
Medical Pathology
Researcher
Verona
Italy
1990/1993
USL 31, District of
Mantova City
Full-time res.ass.
Mantova
Italy
1993/2005
Institute of Hematology
and Medical Oncology
"Seràgnoli"
Medical Doctor
University of Bologna,
Bologna
Italy
2005/2006
Institute of Hematology
and Medical Oncology
"Seràgnoli"
Associate Professor
University og Bologna,
Bologna
Italy
Codice Riferimento: 4121
Page 25 of 44
PUBLICATIONS
Listed, in chronological order, are title, all authors, and complete reference of all publications 2002-2006.
Principal Investigator's Full Name: Professore Martinelli Giovanni
Authors
Title
Journal
Year
Vol.
Nr.
Pages
IF
Nyakern M,Tazzari PL,Finelli
C,Bosi C,Follo MY,Grafone
T,Piccaluga PP,Martinelli
G,Cocco L,Martelli AM
Frequent elevation of Akt kinase
Leukemia
phosphorylation in blood marrow
and peripheral blood mononuclear
cells from high-risk
myelodysplastic syndrome
patients.
2006 Feb
20
230-8
5,81
Piccaluga PP,Malagola
M,Rondoni M,Ottaviani E,Testoni
N,Laterza C,Visani G,Pileri
SA,Martinelli G,Baccarani M
Poor outcome of adult acute
lymphoblastic leukemia patients
carrying the (1;19)(q23;p13)
translocation.
Leuk Lymphoma
2006 Mar
47
469-72
1,147
Martinelli G,Iacobucci I,Rosti
G,Pane F,Amabile M,Castagnetti
F,Cilloni D,Soverini S,Testoni
N,Specchia G,Merante S,Zaccaria
A,Frassoni F,Saglio G,Baccarani
M
Prediction of response to imatinib
by prospective quantitation of
BCR-ABL transcript in late
chronic phase chronic myeloid
leukemia patients.
Ann Oncol
2006 Mar
17
495-502
4,335
Cavo M,Terragna C,Renzulli
M,Zamagni E,Tosi P,Testoni
N,Nicci C,Cangini D,Tacchetti
P,Grafone T,Cellini C,Ceccolini
M,Perrone G,Martinelli
G,Baccarani M,Guardigni L
Poor outcome with front-line
autologous transplantation in
t(4;14) multiple myeloma: low
complete remission rate and short
duration of remission.
J Clin Oncol
2006 Jan 20
24
e4-5
9,835
Iacobucci I,Rosti G,Amabile
M,Poerio A,Soverini S,Cilloni
D,Testoni N,Abruzzese
E,Montefusco E,Ottaviani
E,Iuliano F,Russo D,Gobbi
M,Alimena G,Martino B,Terragna
C,Pane F,Saglio G,Baccarani
M,Martinelli G
Comparison between patients with J Clin Oncol
Philadelphia-positive chronic
phase chronic myeloid leukemia
who obtained a complete
cytogenetic response within 1 year
of imatinib therapy and those who
achieved such a response after 12
months of treatment.
2006 Jan 20
24
454-9
Merante S,Chichino G,Boveri
E,Gottardi E,Soverini S,Cilloni
D,Martinelli G
First case of an AIDS patient with
systemic mast cell disease
associated with FIP1-positive
eosinophilia treated with imatinib
mesylate therapy.
2006 Feb 1
24
e6-7
Codice Riferimento: 4121
J Clin Oncol
F/L
FC/CA
Relevant
X
First
corr-author
X
9,835
Last
corr-author
X
9,835
Last
corr-author
Page 26 of 44
Authors
Title
Journal
Year
Vol.
Nr.
Pages
IF
9,782
Hughes T,Deininger M,Hochhaus
A,Branford S,Radich J,Kaeda
J,Baccarani M,Cortes J,Cross
NC,Druker BJ,Gabert J,Grimwade
D,Hehlmann R,Kamel-Reid
S,Lipton JH,Longtine J,Martinelli
G,Saglio G,Soverini S,Stock
W,Goldman JM
Monitoring CML patients
Blood
responding to treatment with
tyrosine kinase inhibitors: review
and recommendations for
harmonizing current methodology
for detecting BCR-ABL transcripts
and kinase domain mutations and
for expressing results.
2006 Jul 1
108
28-37
Piccaluga PP,Martinelli
G,Baccarani M
Advances in the treatment for
haematological malignancies.
2006 Apr
7
721-32
Iacobucci I,Saglio G,Rosti
G,Testoni N,Pane F,Amabile
M,Poerio A,Soverini S,Bassi
S,Cilloni D,Bassan R,Breccia
M,Lauria F,Izzo B,Merante
S,Frassoni F,Paolini S,Montefusco
E,Baccarani M,Martinelli
G,GIMEMA Working Party on
Chronic Myeloid Leukemia
Achieving a major molecular
Clin Cancer Res
response at the time of a complete
cytogenetic response (CCgR)
predicts a better duration of CCgR
in imatinib-treated chronic
myeloid leukemia patients.
2006 May 15
12
3037-42
Potenza L,Luppi M,Riva
G,Ottaviani E,Zucchini P,Morselli
M,Volzone F,Forghieri
F,Martinelli G,Torelli G
May the correlation between
Kit-D816 mutation and
AML1-ETO level change the use
of prognostic factors in t(8;21)
AML?
Leuk Res
2006 Jun 5
2,244
Bussolari R,Candini O,Colomer
D,Corradini F,Guerzoni C,Mariani
SA,Cattelani S,Silvestri C,Pecorari
L,Iacobucci I,Soverini S,Fasano
T,Martinelli G,Cervantes
F,Calabretta B
Coding sequence and intron-exon
junctions of the c-myb gene are
intact in the chronic phase and
blast crisis stages of chronic
myeloid leukemia patients.
Leuk Res
2006 Jun 22
2,244
Visani G,Olivieri A,Malagola
M,Brunori M,Piccaluga PP,Capelli
D,Pomponio G,Martinelli
G,Isidori A,Sparaventi G,Leoni P
Consolidation therapy for adult
acute myeloid leukemia: a
systematic analysis according to
evidence based medicine.
Leuk Lymphoma
2006 Jun
47
1091-102
1,147
Piccaluga PP,Vigna E,Placci
A,Agostinelli C,Laterza
C,Papayannidis C,Leone
O,Martinelli G,Zinzani
PL,Baccarani M,Pileri SA
Primary cardiac non-Hodgkin
lymphoma presenting with atrial
flutter and pericardial effusion.
Br J Haematol
2006 Aug
134
356
3,195
Codice Riferimento: 4121
Expert Opin
Pharmacother
5,623
F/L
Last
FC/CA
Relevant
fco-author
X
corr-author
X
Page 27 of 44
Authors
Title
Journal
Tagliafico E,Tenedini
E,Manfredini R,Grande A,Ferrari
F,Roncaglia E,Bicciato S,Zini
R,Salati S,Bianchi E,Gemelli
C,Montanari M,Vignudelli
T,Zanocco-Marani T,Parenti
S,Paolucci P,Martinelli
G,Piccaluga PP,Baccarani
M,Specchia G,Torelli U,Ferrari S
Identification of a molecular
signature predictive of sensitivity
to differentiation induction in
acute myeloid leukemia.
Leukemia
Martinelli G,Iacobucci I,Soverini
S,Cilloni D,Saglio G,Pane
F,Baccarani M
Monitoring minimal residual
Hematol Oncol
disease and controlling drug
resistance in chronic myeloid
leukaemia patients in treatment
with imatinib as a guide to clinical
management.
Piccaluga PP,Martinelli
G,Rondoni M,Visani G,Baccarani
M
Advances and potential treatment
for Philadelphia
chromosome-positive adult acute
lymphoid leukaemia.
Expert Opin Biol
Ther
Year
2006 Oct
Vol.
Nr.
20
Pages
IF
1751-8
5,81
2006 Sep 20
2006 Oct
F/L
FC/CA
Relevant
First
corr-author
X
2,446
fco-author
X
2,216
fco-author
fco-author
2,393
6
1011-22
Bianchini M,Martinelli G,Renzulli cDNA microarray study to identify Cancer
M,Gonzalez Cid M,Larripa I
expression changes relevant for
Chemother
apoptosis in K562 cells co-treated Pharmacol
with amifostine and imatinib.
2006 Sep 29
Soverini S,Martinelli G,Colarossi
S,Gnani A,Castagnetti F,Rosti
G,Bosi C,Paolini S,Rondoni
M,Piccaluga PP,Palandri
F,Giannoulia P,Marzocchi
G,Luatti S,Testoni N,Iacobucci
I,Cilloni D,Saglio G,Baccarani M
2006 Nov 20
24
e51-2
9,835
2006 Jan
20
61-7
5,81
Presence or the Emergence of a
F317L BCR-ABL Mutation May
Be Associated With Resistance to
Dasatinib in Philadelphia
Chromosome-Positive Leukemia.
J Clin Oncol
Cilloni D,Messa F,Arruga
The NF-kappaB pathway blockade Leukemia
F,Defilippi I,Morotti A,Messa
by the IKK inhibitor PS1145 can
E,Carturan S,Giugliano E,Pautasso overcome imatinib resistance.
M,Bracco E,Rosso V,Sen
A,Martinelli G,Baccarani
M,Saglio G
Codice Riferimento: 4121
X
X
Page 28 of 44
Authors
Title
Journal
Year
Vol.
Nr.
Pages
IF
F/L
FC/CA
Cappellini A,Mantovani I,Tazzari
PL,Grafone T,Martinelli G,Cocco
L,Martelli AM
Application of flow cytometry to
molecular medicine: detection of
tumor necrosis factor-related
apoptosis-inducing ligand
receptors in acute myeloid
leukaemia blasts.
Int J Mol Med
2005 Dec
16
1041-8
3,19
Piccaluga PP,Martinelli
G,Malagola M,Rondoni
M,Bonifazi F,Bandini G,Visani
G,Baccarani M
Alemtuzumab in the treatment of
relapsed acute lymphoid
leukaemia.
Leukemia
2005 Jan
19
135; author
reply 136
5,81
fco-author
2005 Jan 15
105
904; author
reply 905
9,782
fco-author
Malagola M,Martinelli G,Rondoni Imatinib mesylate in the treatment
M,Paolini S,Gaitani S,Arpinati
of c-kit-positive acute myeloid
M,Piccaluga PP,Amabile M,Basi leukemia: is this the real target?
C,Ottaviani E,Candoni A,Gottardi
E,Cilloni D,Bocchia M,Saglio
G,Lauria F,Fanin R,Visani
G,Marre MC,Maderna M,Rancati
F,Vinaccia V,Russo D,Baccarani
M
Blood
Nicci C,Ottaviani E,Luatti
S,Grafone T,Tonelli M,Motta
MR,Malagola M,Marzocchi
G,Martinelli G,Baccarani
M,Testoni N
Molecular and cytogenetic
characterization of a new case of
t(5;17)(q35;q21) variant acute
promyelocytic leukemia.
Leukemia
2005 Mar
19
470-2
5,81
Pane F,Cimino G,Izzo B,Camera
A,Vitale A,Quintarelli C,Picardi
M,Specchia G,Mancini M,Cuneo
A,Mecucci C,Martinelli G,Saglio
G,Rotoli B,Mandelli F,Salvatore
F,Foa R,GIMEMA group
Significant reduction of the hybrid Leukemia
BCR/ABL transcripts after
induction and consolidation
therapy is a powerful predictor of
treatment response in adult
Philadelphia-positive acute
lymphoblastic leukemia.
2005 Apr
19
628-35
5,81
Martinelli G,Soverini S,Rosti
G,Cilloni D,Baccarani M
New tyrosine kinase inhibitors in
chronic myeloid leukemia.
2005 Apr
90
534-41
4,192
Codice Riferimento: 4121
Haematologica
First
corr-author
Relevant
X
Page 29 of 44
Authors
Title
Journal
Soverini S,Martinelli G,Rosti
G,Bassi S,Amabile M,Poerio
A,Giannini B,Trabacchi
E,Castagnetti F,Testoni N,Luatti
S,de Vivo A,Cilloni D,Izzo B,Fava
M,Abruzzese E,Alberti D,Pane
F,Saglio G,Baccarani M
ABL mutations in late chronic
phase chronic myeloid leukemia
patients with up-front cytogenetic
resistance to imatinib are
associated with a greater
likelihood of progression to blast
crisis and shorter survival: a study
by the GIMEMA Working Party
on Chronic Myeloid Leukemia.
J Clin Oncol
Martinelli G,Soverini S,Rosti
G,Baccarani M
Dual tyrosine kinase inhibitors in
chronic myeloid leukemia.
Leukemia
Russo D,Malagola M,de Vivo
A,Fiacchini M,Martinelli
G,Piccaluga PP,Damiani
D,Candoni A,Michielutti
A,Castelli M,Testoni N,Ottaviani
E,Rondoni M,Pricolo G,Mazza
P,Zuffa E,Zaccaria A,Raspadori
D,Bocchia M,Lauria F,Bonini
A,Avanzini P,Gugliotta L,Visani
G,Fanin R,Baccarani M
Year
Vol.
Nr.
Pages
IF
2005 Jun 20
23
4100-9
9,835
2005 Nov
19
1872-9
5,81
Multicentre phase III trial on
Br J Haematol
fludarabine, cytarabine (Ara-C),
and idarubicin versus idarubicin,
Ara-C and etoposide for induction
treatment of younger, newly
diagnosed acute myeloid
leukaemia patients.
2005 Oct
131
172-9
3,195
Buonamici S,Ottaviani E,Visani
G,Bonifazi F,Fiacchini
M,Baccarani M,Martinelli G
Patterns of AML1-ETO transcript
expression in patients with acute
myeloid leukemia and t(8;21) in
complete hematologic remission.
Haematologica
2004 Jan
89
103-5
4,192
Martinelli G,Malagola
M,Ottaviani E,Rosti G,Trabacchi
E,Baccarani M
Imatinib mesylate can induce
complete molecular remission in
FIP1L1-PDGFR-a positive
idiopathic hypereosinophilic
syndrome.
Haematologica
2004 Feb
89
236-7
4,192
Piccaluga PP,Poletti G,Martinelli
G,Gherlinzoni F
Babesia infection in Italy.
Lancet Infect Dis
2004 Apr
4
212
10,788
Martinelli G,Rondoni
M,Buonamici S,Ottaviani
E,Piccaluga PP,Malagola
M,Baccarani M
Molecular monitoring to identify a Haematologica
threshold of CBFbeta/MYH11
transcript below which continuous
complete remission of acute
myeloid leukemia inv16 is likely.
2004 Apr
89
495-7
Dose increase of imatinib mesylate Eur J Haematol
may overcome acquired resistance
in bcr/abl-positive acute lymphoid
leukaemia.
2004 Apr
72
302-3
Piccaluga PP,Malagola
M,Rondoni M,Amabile M,Paolini
S,Soverini S,Gaitani S,Visani
Codice
Riferimento:
4121 G
G,Baccarani
M,Martinelli
F/L
First
FC/CA
Relevant
fco-author
X
corr-author
X
corr-author
First
corr-author
4,192
First
corr-author
1,729
Last
corr-author
Page 30 of 44
Authors
Title
Journal
Soverini S,Martinelli G,Amabile
M,Poerio A,Bianchini M,Rosti
G,Pane F,Saglio G,Baccarani
M,Italian Cooerative Study Group
on Chronic Myeloid
Leukemia,European
LeukemiaNet-6th Framework
Program of the European
Community
Denaturing-HPLC-based assay for Clin Chem
detection of ABL mutations in
chronic myeloid leukemia patients
resistant to Imatinib.
Piccaluga PP,Luatti S,Ascani
S,Bianchini M,Malagola
M,Rondoni M,Gaitani S,Testoni
N,Pileri SA,Baccarani
M,Martinelli G,Visani G
Identification of a novel
t(1;9)(q11;q34) in acute
myelocytic leukemia.
Piccaluga PP,Ricci P,Martinelli
G,Malagola M,Rondoni M,Visani
G
Year
Vol.
Nr.
Pages
IF
F/L
FC/CA
Relevant
fco-author
X
2004 Jul
50
1205-13
6,501
Cancer Genet
Cytogenet
2004 May
151
85-6
1,577
Prompt resolution of nasal
aspergillosis with intranasal
instillation of liposomal
amphotericin-B (amBisome) and
granulocyte transfusions.
Leuk Lymphoma
2004 Mar
45
637-8
1,147
Piccaluga PP,Martinelli
G,Malagola M,Rondoni
M,Bianchini M,Vigna E,Bosi
C,Gaitani S,Visani G,Baccarani M
Anti-leukemic and anti-GVHD
effects of campath-1H in acute
lymphoblastic leukemia relapsed
after stem-cell transplantation.
Leuk Lymphoma
2004 Apr
45
731-3
1,147
fco-author
Piccaluga PP,Martinelli
G,Rondoni M,Malagola M,Gaitani
S,Isidori A,Bonini A,Gugliotta
L,Luppi M,Morselli M,Sparaventi
G,Visani G,Baccarani M
Gemtuzumab ozogamicin for
relapsed and refractory acute
myeloid leukemia and myeloid
sarcomas.
Leuk Lymphoma
2004 Sep
45
1791-5
1,147
fco-author
2004 Sep
28
987-90
2,244
fco-author
2004 Nov 15
104
3126-35
9,782
fco-author
Piccaluga PP,Martinelli
First experience with gemtuzumab Leuk Res
G,Rondoni M,Malagola M,Gaitani ozogamicin plus cytarabine as
S,Visani G,Baccarani M
continuous infusion for elderly
acute myeloid leukaemia patients.
Tenedini E,Fagioli ME,Vianelli
N,Tazzari PL,Ricci F,Tagliafico
E,Ricci P,Gugliotta L,Martinelli
G,Tura S,Baccarani M,Ferrari
S,Catani L
Codice Riferimento: 4121
Gene expression profiling of
normal and malignant
CD34-derived megakaryocytic
cells.
Blood
Page 31 of 44
Authors
Title
Pruneri G,Valentini S,Fabris S,Del
Curto B,Laszlo D,Bertolini
F,Martinelli G,Leocata P,Viale
G,Neri A
Cyclin D3 immunoreactivity in
follicular lymphoma is
independent of the
t(6;14)(p21.1;q32.3) translocation
or cyclin D3 gene amplification
and is correlated with histologic
grade and Ki-67 labeling index.
Baccarani M,Martinelli G,Rosti
G,Trabacchi E,Testoni N,Bassi
S,Amabile M,Soverini
S,Castagnetti F,Cilloni D,Izzo
B,de Vivo A,Messa E,Bonifazi
F,Poerio A,Luatti S,Giugliano
E,Alberti D,Fincato G,Russo
D,Pane F,Saglio G,GIMEMA
Working Party on Chronic
Myeloid Leukemia
Imatinib and pegylated human
Blood
recombinant interferon-alpha2b in
early chronic-phase chronic
myeloid leukemia.
Tazzari PL,Cappellini A,Grafone
T,Mantovani I,Ricci F,Billi
AM,Ottaviani E,Conte
R,Martinelli G,Martelli AM
Detection of serine 473
phosphorylated Akt in acute
myeloid leukaemia blasts by flow
cytometry.
Malagola M,Martinelli G,Rondoni Soft tissue and skeletal
M,Ottaviani E,Piccaluga PP,Ricci involvement in
P,Visani G,Baccarani M
FIP1L1-PDGFR-alpha positive
chronic eosinophilic leukemia:
imatinib mesylate may induce
complete molecular and imaging
remission.
Journal
Int J Cancer
Year
Vol.
Nr.
Pages
IF
2004 Oct 20
112
71-7
4,416
2004 Dec 15
104
4245-51
9,782
Br J Haematol
2004 Sep
126
675-81
3,195
Haematologica
2004 Aug
89
ECR25
4,192
Visani G,Isidori A,Grafone T,Tosi
P,Santini V,Malagola M,Martinelli
G,Piccaluga PP,Gaziev
D,Ottaviani E,Sparaventi G,Tura S
No preferential sensitivity of
Leuk Lymphoma
t(8;21) acute myeloid leukemias to
cytosine arabinoside in vitro: is
intensity of therapy or high dose
Ara-C crucial for response?
2004 Jul
45
1361-4
1,147
Piccaluga PP,Malagola
M,Amabile M,Rondoni M,Paolini
S,Pane F,Russo D,Visani
G,Baccarani M,Martinelli G
The achievement of molecular
complete remission during
treatment with imatinib mesylate
correlates with relapse-free
survival in bcr/abl-positive acute
lymphoid leukemia patients.
2004 Oct
89
1269-71
4,192
Codice Riferimento: 4121
Haematologica
F/L
FC/CA
Relevant
fco-author
X
fco-author
Last
corr-author
Page 32 of 44
Authors
Title
Journal
Rosti G,Martinelli G,Bassi
S,Amabile M,Trabacchi
E,Giannini B,Cilloni D,Izzo B,De
Vivo A,Testoni N,Cambrin
GR,Bonifazi F,Soverini S,Luatti
S,Gottardi E,Alberti D,Pane
F,Salvatore F,Saglio G,Baccarani
M,Study Committee, Italian
Cooperative Study Group for
Chronic Myeloid
Leukemia,Writing Committee,
Italian Cooperative Study Group
for Chronic Myeloid Leukemia
Molecular response to imatinib in Blood
late chronic-phase chronic myeloid
leukemia.
Mancini M,Brusa G,Benvenuti
M,Mazzacurati L,Campanini
F,Barbieri E,Cammelli S,Calonghi
N,Martinelli G,Baccarani
M,Santucci MA
The p210BCR-ABL tyrosine
kinase of chronic myeloid
leukemia causes resistance to
radio-induced apoptotic death by
inhibiting the proapoptotic BAX
gene.
Piccaluga PP,Martinelli
G,Rondoni M,Malagola
M,Ronconi S,Visani G,Baccarani
M
Low dose gemtuzumab
ozogamicin for relapsed acute
myeloid leukaemia in elderly.
Year
Vol.
Nr.
Pages
IF
F/L
FC/CA
2004 Mar 15
103
2284-90
9,782
Leukemia
2004 Feb
18
370-2
5,81
Haematologica
2003 Dec
88
ECR37
4,192
Martinelli G,Piccaluga PP,Lo
Coco F
FLT3 inhibition as tailored therapy Haematologica
for acute myeloid leukemia.
2003 Jan
88
4-8
4,192
First
corr-author
Baccarani M,Russo D,Rosti
G,Martinelli G
Interferon-alfa for chronic myeloid Semin Hematol
leukemia.
2003 Jan
40
22-33
3,835
Last
corr-author
Leukemia
2003 Mar
17
554-9
5,81
Molecular monitoring of acute
Leukemia
myeloid leukemia associated with
inv(16): threshold of
CBFbeta/MYH11 transcript copy
number above which relapse
occurs and below which
continuous Complete Remission is
likely.
2003 Mar
17
650-1; author
reply 651-2
5,81
First
corr-author
Rosti G,Bonifazi F,Trabacchi
A phase II study of
E,De Vivo A,Bassi S,Martinelli
alpha-interferon and oral
G,Testoni N,Russo D,Baccarani M arabinosyl cytosine (YNK01) in
chronic myeloid leukemia.
Martinelli G,Buonamici S,Visani
G,Malagola M,Piccaluga
PP,Isidori A,Bosi C,Bonifazi
F,Soverini S,Terragna C,Amabile
M,Giannini B,Baccarani M
Codice Riferimento: 4121
Relevant
fco-author
X
fco-author
Page 33 of 44
Authors
Kroger N,Zander AR,Martinelli
G,Ferrante P,Moraleda JM,Da
Prada GA,Demirer T,Socie
G,Rosti G,European Group for
Blood and Marrow
Transplantation
Title
Journal
Vol.
Nr.
Pages
IF
F/L
FC/CA
2003 Apr
14
554-8
4,335
Rosti G,Trabacchi E,Bassi
Risk and early cytogenetic
Haematologica
S,Bonifazi F,de Vivo A,Martinelli response to imatinib and interferon
G,Alberti D,Fincato G,Saglio
in chronic myeloid leukemia.
G,Baccarani M,Italian Cooperative
Study Group on Chronic Myeloid
Leukemia
2003 Mar
88
256-9
4,192
Visani G,Piccaluga PP,Martinelli Sustained molecular remission in
G,Rossi M,Malagola M,Baccarani advanced acute promyelocytic
M
leukemia with combined pulsed
retinoic acid and arsenic trioxide.
Clinical evidence of synergistic
effect and real-time quantification
of minimal residual disease.
Haematologica
2003 Apr
88
ELT15
4,192
Martinelli G,Agazzi A,Laszlo
D,Santoro P,Mancuso P,Pruneri
GC,Greco P,Bertolini F
Idarubicin containing regimen in
multiple myeloma: preliminary
results of a pilot study using a
modified "TANDEM" transplant
program.
Leuk Lymphoma
2003 Feb
44
299-302
1,147
First
corr-author
Soverini S,Cavo M,Cellini
C,Terragna C,Zamagni E,Ruggeri
D,Testoni N,Tosi P,De Vivo
A,Amabile M,Grafone T,Ottaviani
E,Giannini B,Cangini D,Bonifazi
F,Neri A,Fabris S,Tura
S,Baccarani M,Martinelli G
Cyclin D1 overexpression is a
favorable prognostic variable for
newly diagnosed multiple
myeloma patients treated with
high-dose chemotherapy and
single or double autologous
transplantation.
Blood
2003 Sep 1
102
1588-94
9,782
Last
corr-author
Codice Riferimento: 4121
Low incidence of secondary
Ann Oncol
myelodysplasia and acute myeloid
leukemia after high-dose
chemotherapy as adjuvant therapy
for breast cancer patients: a study
by the Solid Tumors Working
Party of the European Group for
Blood and Marrow
Transplantation.
Year
Relevant
Page 34 of 44
Authors
Title
Journal
Corradini P,Cavo M,Lokhorst
H,Martinelli G,Terragna
C,Majolino I,Valagussa
P,Boccadoro M,Samson
D,Bacigalupo A,Russell
N,Montefusco V,Voena C,Gahrton
G,Chronic Leukemia Working
Party of the European Group for
Blood and Marrow
Transplantation (EBMT)
Molecular remission after
Blood
myeloablative allogeneic stem cell
transplantation predicts a better
relapse-free survival in patients
with multiple myeloma.
Rosti G,Testoni N,Martinelli
G,Baccarani M
The cytogenetic response as a
surrogate marker of survival.
Year
Vol.
Nr.
Pages
IF
F/L
FC/CA
2003 Sep 1
102
1927-9
9,782
Semin Hematol
2003 Apr
40
56-61
3,835
Brusa G,Benvenuti M,Mazzacurati
L,Mancini M,Pattacini
L,Martinelli G,Barbieri
E,Greenberger JS,Baccarani
M,Santucci MA
p53 loss of function enhances
Haematologica
genomic instability and accelerates
clonal evolution of murine
myeloid progenitors expressing the
p(210)BCR-ABL tyrosine kinase.
2003 Jun
88
622-30
4,192
Piccaluga PP,Martinelli
G,Malagola M,Rondoni
M,Bianchini M,Visani
G,Baccarani M
Complete remission in acute
Haematologica
myeloid leukemia with
granulocyte-colony stimulating
factor without chemotherapy.
Report of cytogenetic remission of
a t(9;11)(p22q23) positive AML
patient and review of literature.
2003 Aug
88
ECR28
4,192
Bianchini M,Ottaviani E,Grafone
T,Giannini B,Soverini S,Terragna
C,Amabile M,Piccaluga
PP,Malagola M,Rondoni M,Bosi
C,Baccarani M,Martinelli G
Rapid detection of Flt3 mutations Clin Chem
in acute myeloid leukemia patients
by denaturing HPLC.
2003 Oct
49
1642-50
6,501
Last
corr-author
Piccaluga PP,Bianchini
M,Martinelli G
Novel FLT3 point mutation in
acute myeloid leukaemia.
2003 Oct
4
604
7,47
Last
corr-author
2003 Nov
88
1221-8
4,192
First
corr-author
Lancet Oncol
Martinelli G,Ottaviani
Association of 3q21q26 syndrome Haematologica
E,Buonamici S,Isidori A,Borsaru with different RPN1/EVI1 fusion
G,Visani G,Piccaluga PP,Malagola transcripts.
M,Testoni N,Rondoni M,Nucifora
G,Tura S,Baccarani M
Codice Riferimento: 4121
Relevant
fco-author
Page 35 of 44
Authors
Title
Journal
Year
Vol.
Nr.
Pages
IF
F/L
FC/CA
Last
corr-author
Buonamici S,Ottaviani E,Testoni
N,Montefusco V,Visani
G,Bonifazi F,Amabile M,Terragna
C,Ruggeri D,Piccaluga PP,Isidori
A,Malagola M,Baccarani M,Tura
S,Martinelli G
Real-time quantitation of minimal Blood
residual disease in inv(16)-positive
acute myeloid leukemia may
indicate risk for clinical relapse
and may identify patients in a
curable state.
2002 Jan 15
99
443-9
9,782
Baccarani M,Rosti G,de Vivo
A,Bonifazi F,Russo D,Martinelli
G,Testoni N,Amabile M,Fiacchini
M,Montefusco E,Saglio G,Tura
S,Italian Cooperative Study Group
on Myeloid Leukemia
A randomized study of
interferon-alpha versus
interferon-alpha and low-dose
arabinosyl cytosine in chronic
myeloid leukemia.
Blood
2002 Mar 1
99
1527-35
9,782
Martinelli G,Setola E,Ricci P
Bone marrow of a patient with
acute non-lymphoblastic
leukaemia associated with the
t(1;7)(p11;p11) chromosomal
translocation.
Haematologica
2002 Mar
87
EIM08
4,192
First
corr-author
Martinelli G,Ottaviani
Two more inv(16) acute myeloid
E,Buonamici S,Isidori A,Malagola leukemia cases with infrequent
M,Piccaluga P,Baccarani M
CBFbeta-MYH11 fusion
transcript: clinical and molecular
findings.
Haematologica
2002 May
87
554-5
4,192
First
corr-author
Martinelli G,Amabile M,Giannini Novel types of bcr-abl transcript
B,Terragna C,Ottaviani E,Soverini with breakpoints in BCR exon 8
S,Saglio G,Rosti G,Baccarani M
found in Philadelphia positive
patients with typical chronic
myeloid leukemia retain the
sequence encoding for the DBLand CDC24 homology domains
but not the pleckstrin homology
one.
Haematologica
2002 Jul
87
688-94;
discussion 694
4,192
First
corr-author
Piccaluga PP,Visani G,Pileri
SA,Ascani S,Grafone T,Isidori
A,Malagola M,Finelli C,Martinelli
G,Ricci P,Baccarani M,Tura S
Clinical efficacy and
Leukemia
antiangiogenic activity of
thalidomide in myelofibrosis with
myeloid metaplasia. A pilot study.
2002 Sep
16
1609-14
5,81
Catani L,Vianelli N,Amabile
M,Pattacini L,Valdre L,Fagioli
ME,Poli M,Gugliotta L,Moi
P,Marini MG,Martinelli G,Tura
S,Baccarani M
Nuclear factor-erythroid 2
Leukemia
(NF-E2) expression in normal and
malignant megakaryocytopoiesis.
2002 Sep
16
1773-81
5,81
Codice Riferimento: 4121
Relevant
Page 36 of 44
Authors
Title
Journal
Year
Vol.
Nr.
Pages
IF
Piccaluga PP,Visani G,Martinelli
G,Isidori A,Malagola M,Rondoni
M,Baccarani M,Tura S
Liposomal daunorubicin
(DaunoXome) for treatment of
relapsed meningeal acute myeloid
leukemia.
Leukemia
2002 Sep
16
1880-1
5,81
Visani G,Isidori A,Malagola
M,Alberti D,Capdeville
R,Martinelli G,Piccaluga
PP,Amabile M,Guiducci B,Tura
S,Baccarani M
Efficacy of imatinib mesylate
(STI571) in conjunction with
alpha-interferon: long-term
quantitative molecular remission
in relapsed
P-190(BCR-ABL)-positive acute
lymphoblastic leukemia.
Leukemia
2002 Oct
16
2159-60
5,81
Soverini S,Terragna C,Testoni
N,Ruggeri D,Tosi P,Zamagni
E,Cellini C,Cavo M,Baccarani
M,Tura S,Martinelli G
Novel mutation and RNA splice
variant of fibroblast growth factor
receptor 3 in multiple myeloma
patients at diagnosis.
Haematologica
2002 Oct
87
1036-40
4,192
Zaccaria A,Valenti A,Toschi
M,Salvucci M,Cipriani
R,Ottaviani E,Martinelli G
Cryptic translocation of
PML/RARA on 17q. A rare event
in acute promyelocytic leukemia.
Cancer Genet
Cytogenet
2002 Oct 15
138
169-73
1,577
Codice Riferimento: 4121
F/L
FC/CA
Last
corr-author
Relevant
Page 37 of 44
EXISTING/PENDING SUPPORT
Funding Agency
AIRC
Codice Riferimento: 4121
Project Title
From
To
Amount
Tyrosine-kinases as molecular target of the treatment
of Ph+ chronic and acute myeloid leukemias
2004
2004
35.000
Page 38 of 44
EXISTING/PENDING SUPPORT JUSTIFICATION
EXISTING/PENDING SUPPORT
Only for year 2004 AIRC project:
TYROSINE‐KINASES AS MOLECULAR TARGET OF THE TREATMENT OF Ph+ CHRONIC AND ACUTE MYELOID LEUKEMIAS
Codice Riferimento: 4121
Page 39 of 44
UNDESIRABLE REVIEWERS
Codice Riferimento: 4121
Page 40 of 44
BIO-ETHICAL REQUIREMENTS
Does the proposed research involve:
HUMAN EXPERIMENTATION
YES
X
NO
Human experimentation includes involvement of human subjects and other issues with ethical implications.
If yes include clearance from the competent ethical committee (as addendum E). If approval of the ethical committee is not available at
moment of submission, include as addendum E, a signed statement from the proponent pledging to obtain it before the start of the
research. AIRC will not erogate funds in the absence of an ethical committee clearance.
ANIMAL EXPERIMENTATION
YES
NO
X
If yes the following statement will have to be signed in the paper submission:
The Ethical Committee for animal use in cancer research has evaluated the proposal.
The Committee considers that the preliminary results, obtained in in-vitro experiments deserve a preclinical study.
The procedures related to animal use are accurately described in the proposal and conform to all regulations protecting animals used for
research purposes, including those of the DL 116/92. The experiments described in the proposal will be performed following the detailed
Internal Regulation drawn-up according to: Workamn P., et al. (1998) United Kingdom Coordinating Committee on Cancer Research
(Guidelines for the welfare of animals in experimental neoplasia. Br. J. Cancer 77: 1-10).
Date:
15/12/2006
Codice Riferimento: 4121
Name of PI
Professore Martinelli
Giovanni
Signature
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RESEARCH INTERRUPTIONS AND JUSTIFICATION
Codice Riferimento: 4121
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ADDENDA
Addendum A: Letter of the NUSUG sponsor (uploaded as PDF file)
Addendum B: Formal letters of collaboration (uploaded as PDF file)
Addendum C: Institutional letter indirect costs/overhead (uploaded as PDF file)
Addendum D: Clearance from ethical committee (uploaded as PDF file)
Codice Riferimento: 4121
;
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ADDENDA D - Clearance
ALMA MATER STUDIORUM
UNIVERSITÀ DI BOLOGNA
S.S.R. EMILIA-ROMAGNA
Azienda Ospedaliero – Universitaria di Bologna
Policlinico S. Orsola-Malpighi
Dipartimento di Ematologia, Oncologia
e Medicina di Laboratorio
Istituto di Ematologia e Oncologia Medica
“Lorenzo e Ariosto Seràgnoli”
Prof. M. Baccarani, Direttore
Dr. M. Fiacchini
Prof. M. Cavo
Dr. G. Bandini
Dr.ssa M.R. Motta
Dr. G. Rosti
Dr. C. Finelli
Dr.ssa N. Testoni
Dr.ssa S. Rizzi
Prof. P.L. Zinzani
Dr. N. Vianelli
Prof. R.M. Lemoli
Dr.ssa P. Tosi
Prof. G. Martinelli
Dr.ssa L. Catani
Dr. D. Rondelli
Dr.ssa E. Ottaviani
Dr. A. De Vivo
Dr.ssa F. Bonifazi
Dr. M. Arpinati
Dr.ssa M. Stanzani
Dr. Antonio Curti
Bologna, 29 novembre 2006
Spettabile AIRC
Via Corridoni, 7
20122 MILANO
Rif.: AIRC – Grant Proposal 2007 – Addendum D – clearance from ethical committee.
This study must be carried out in compliance with the principles of Good Clinical Practice, as
described in Institution Etical Board (IEB) standard operating procedures and:
ICH
Harmonized
Tripartite
Guidelines
for
Good
Clinical
Practice
1996.
Directive 91/507/EEC, The Rules Governing Medicinal Products in the European Community.
US 21 Code of Federal Regulations dealing with clinical studies (including parts 50 and 56
concerning informed consent and IRB regulations).
Declaration of Helsinki and amendments, concerning medical research in humans
(Recommendations Guiding Physicians in Biomedical Research Involving Human Subjects).
The principal investigator agrees when signing the study to adhere to the instructions and
procedures described in it and thereby to adhere to the principles of Good Clinical Practice that it
conforms to.
This study will be submitted to the approval of Ethical Committee of Azienda Ospedaliera di
Bologna.
Dr. Giovanni Martinelli
Via Massarenti, 9 – 40138 Bologna
Portineria...................................................................................... tel 0516363680 / 0516363700 / 0516363800 - fax 0516364037
Segreteria Direzione .................................................................... tel 051390413 - fax 051398973 - e-mail [email protected]
Segreteria Ambulatorio ............................................................................................tel 0516363480 - e-mail [email protected]
a
Capo Sala Reparto Degenza I Sezione ......................................................................tel 0516364081 - e-mail [email protected]
a
Capo Sala Reparto Degenza II Sezione “Semi Intensivo” ..........................................tel 0516364090 - e-mail [email protected]
a
Capo Sala Reparto Degenza III Sezione “BCM”.........................................................tel 0516363479 - e-mail [email protected]
Capo Sala Day Hospital.........................................................................................tel 0516363815 - e-mail [email protected]
fz
Codice Riferimento: 4121
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