New molecular markers to evaluate susceptibility and prognosis and to identify targeted-therapies in adult and childhood acute leukemia, de novo or secondary.

Research project

Description

This projects is the result of the collaboration of 9 centers, from 7 Universities, which have been involved for several years in the treatment of childhood and adult acute myeloid leukemia, de novo or therapy-related. These centers coordinate or are active members of national and European multicenter clinical trials, for the treatment of children (AIEOP) and adults (GIMEMA,EORTC, European Leukemia Network) and have the expertise and the technologies to identify new genetic and molecular lesions, defining de novo and secondary acute leukemias. To evaluate therapy-related forms, in addition to the Gimema and AIEOP registries, we have a “Secondary leukemia” registry, coordinated by the Department of Hematology of the Universita’ Cattolica, which will be implemented by the collaboration of the 9 groups.
In this research project, we specifically aim at exploting the following tasks for acute lymphoblastic leukemia (ALL):
1. Identification in childhood and adult ALL, of new molecular lesi,ons, particularly of IKZF1 and CRLF2, using quantitative PCR and molecular cytogenetics
a) Evaluation of the impact of the IKZF1 deletion and the potential application of this marker in a prognostic algorithm, to identify patients at high-risk for relapse;
b) Quantification of CRLF2 expression using quantitative PCR and FISH, for the presence of the CRLF2-P2RY8 fusion gene, and characterization of JAK2 and CRLF2 mutations.
2. Evaluation of microRNA , small nucleolar RNA (snoRNA)and RNA-seq to study the transcriptional regulation in ALL
3. Implementation of a new test, known as Combined-in situ fluorescence hybridization (CI-FISH), to be used to identify known genetic alterations and characterize new genetic lesions, important for the pathogenesis of B-ALL, involving MYC and SUPT3H, NUP98 and NUP214, in T-ALL.
In secondary leukemias, we aim at the
1. Expansion of therapy-regated epidemiological registry, creating an international network, involving europena and American centers. The, aims of the registry are to evaluated in a lerge number of patients (expected 600 patients in 3 years): incidence and differential diagnosis between true t-AML, and leukemias secondary to a previous malignancy, treated with surgery alone, to autoimmune disorders or multiple sclerosis, latency between the primary disease and t-AL and role of different chemo- and radiotherapy regimens and schedules.
2. Implementation of molecular tests in the laboratories of participating centers. In particular we will study therapy-related leukemia following ALL, where we have biological constitutional material and the appropriate controls. These studies will allow to evaluate a) susceptibility, by the Affymetrix Genome-Wide Human SNP Array 6., which study constitutional single nucleotide polymorphisms (SNPs) genome-wide ; b) the t-AL genetic profile, by Array-CGH (Comparative Genomic Hybridization)and RNA-seq; c) changes in PTEN expression by RT-PCR and Western-Blot, or tyrosin-kinases (FLT3, RAS, Ja,k-2, ATM) mutations by sequencing; epigenetic changes using HELP methylation arrays, based on digestion of DNA by methylation-sensitive enzymes, as HpaII or MspI.

Expected results: the use of advanced diagnostic technologies gives the basis the definition of refined risk-groups in de novo acute lymphoblastic leukemia, allowing the reduction of dosis and toxicity of classical chemotherapy drugs in favourable risk groups, and the identification of subgroups requiring alternative treatment strategies. In secondary AL, we aim at the identification of a reliable diagnostic test, which allows the definition of a risk-profile patient- and drug-specific. The collaboration of the 9 research groups with outstanding clinical, epidemiological and molecular expertise, will be the ideal ground for the success of this project.
StatusActive
Effective start/end date1/1/11 → …

Funding

  • MIUR - FIRB

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Precursor Cell Lymphoblastic Leukemia-Lymphoma
Leukemia
Registries
Polymerase Chain Reaction
Single Nucleotide Polymorphism
RNA
Technology
Drug Therapy
Small Nucleolar RNA
Social Identification
Comparative Genomic Hybridization
Hematology
DNA Methylation
Fluorescence In Situ Hybridization
MicroRNAs
Routine Diagnostic Tests
Acute Myeloid Leukemia
Epigenomics
Methylation
Multicenter Studies