The goal of this proposal is to identify the genetic basis for polycythemia vera (PV), essential thrombocythemia (ET) and agnogenic myeloid metaplasia (AMM), and to develop more effective treatments or these disorders based on genetic insights into disease pathogenesis. We hypothesize that PV, ET and AMM are due to activating mutations in tyrosine kinases. The hypothesis is based on the observation that the majority of myeloproliferative disorders (MPDs) characterized thus far are caused by activating mutations n tyrosine kinases, including chronic myelogenous leukemia, hypereosinophilic syndrome, and chronic myelomonocytic leukemia. Despite these examples of advances in our understanding of molecular pathogenesis of MPD, the genetic cause of PV, ET or AMM is completely unknown, due in part to rarity of the diseases, lack of heritability, and lack of cytogenetic clues to the identity of the mutant kinases.
The Specific Aims of this proposal are to characterize the role of tyrosine kinase activation in the pathogenesis of PV, ET and AMM. We will use state-of-the-art genome wide screens to discover novel tyrosine kinase mutations, characterize the functional consequences of mutations, and develop specific inhibitors to mutant kinases as potential therapeutic agents. Because these diseases are relatively rare, a key to success of the proposal is acquisition of blood and buccal mucosal swabs from a large cohort of patients which has been successfully achieved. As an exciting proof-of-principle for this approach, we have recently identified JAK2V617Fmutations in most patients with PV and in some patients with ET/AMM.
The Specific Aims are: 1. To use DNA sequence analysis to identify mutations in conserved domains of tyrosine kinases. 2. To use a custom designed oligonucleotide CGH array to discover intersitial deletions that activate tyrosine kinases. 3. Functional characterization of JAK2V617F and other novel genomic events leading to tyrosine kinase activation using in vitro and in vivo assays. 4. Develop and test kinase inhibitors against JAK2V617F and other activated tyrosine kinase using in vitro and in vivo assays. Although beyond the scope of this K08 proposal, the long term goal of this research is to expand the paradigm of tyrosine kinase inhibition as safe and effective therapy for cancer to include the myeloproliferative diseases PV, ET and AMM. We plan to develop preclinical data that will provide a platform for initiating clinical trials with novel kinase inhibitors. (End 0f Abstract)

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08HL082677-06
Application #
7928212
Study Section
Special Emphasis Panel (ZHL1-CSR-O (M1))
Program Officer
Mondoro, Traci
Project Start
2006-08-21
Project End
2011-07-31
Budget Start
2010-08-01
Budget End
2011-07-31
Support Year
6
Fiscal Year
2010
Total Cost
$133,920
Indirect Cost
Name
Sloan-Kettering Institute for Cancer Research
Department
Type
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10065
Lin, X; Rice, K L; Buzzai, M et al. (2013) miR-433 is aberrantly expressed in myeloproliferative neoplasms and suppresses hematopoietic cell growth and differentiation. Leukemia 27:344-52
Berkofsky-Fessler, Windy; Buzzai, Monica; Kim, Marianne K-H et al. (2010) Transcriptional profiling of polycythemia vera identifies gene expression patterns both dependent and independent from the action of JAK2V617F. Clin Cancer Res 16:4339-52
Abdel-Wahab, Omar; Manshouri, Taghi; Patel, Jay et al. (2010) Genetic analysis of transforming events that convert chronic myeloproliferative neoplasms to leukemias. Cancer Res 70:447-52
Busque, Lambert; Paquette, Yves; Provost, Sylvie et al. (2009) Skewing of X-inactivation ratios in blood cells of aging women is confirmed by independent methodologies. Blood 113:3472-4
Abdel-Wahab, Omar; Mullally, Ann; Hedvat, Cyrus et al. (2009) Genetic characterization of TET1, TET2, and TET3 alterations in myeloid malignancies. Blood 114:144-7
Tefferi, A; Pardanani, A; Lim, K-H et al. (2009) TET2 mutations and their clinical correlates in polycythemia vera, essential thrombocythemia and myelofibrosis. Leukemia 23:905-11
Bernard, L; Belisle, C; Mollica, L et al. (2009) Telomere length is severely and similarly reduced in JAK2V617F-positive and -negative myeloproliferative neoplasms. Leukemia 23:287-91
Loriaux, Marc M; Levine, Ross L; Tyner, Jeffrey W et al. (2008) High-throughput sequence analysis of the tyrosine kinome in acute myeloid leukemia. Blood 111:4788-96
Pikman, Yana; Levine, Ross L (2007) Advances in the molecular characterization of Philadelphia-negative chronic myeloproliferative disorders. Curr Opin Oncol 19:628-34
Frohling, Stefan; Scholl, Claudia; Levine, Ross L et al. (2007) Identification of driver and passenger mutations of FLT3 by high-throughput DNA sequence analysis and functional assessment of candidate alleles. Cancer Cell 12:501-13