Chronic myeloid leukemia (CML) is a hematopoietic malignancy characterized by a (9;22) translocation, which generates the constitutively active BCR-ABL tyrosine kinase. BCR-ABL is the critical mediator of disease pathogenesis, exhibiting constitutive kinase activity that drives survival and proliferation through multiple downstream pathways. First-line therapy with the ABL kinase inhibitor imatinib leads to durable responses in the majority of patients with CML, although a substantial number of patients develop resistance to therapy. The second-generation ABL inhibitors nilotinib and dasatinib have proven largely effective as salvage therapies in this clinical scenario. However, the """"""""gatekeeper"""""""" BCR-ABLT315I mutant is insensitive to all three approved drugs, and new ABLT315I inhibitors are in development. Recent clinical reports suggest that sequential ABL tyrosine kinase inhibitor therapy may select for BCR-ABLT315I-inclusive compound mutants (two mutations within the same BCR-ABL molecule) that confer high-level resistance. A cell-based screen will be used to identify BCR-ABL compound mutations that confer resistance to ABLT315I inhibitors, and strategies to minimize emergence of compound mutants will be explored. A second subset of patients exhibits disease progression via a BCR-ABL-independent, """"""""oncogene switching"""""""" mechanism, and the molecular events leading to this phenomenon remain unidentified. To better understand this process, the combination of functional screens using panels of siRNA and kinase inhibitors with high-throughput sequencing and microarray analysis will accelerate the identification of molecular mechanisms of BCR-ABL-independent resistance. To gain a more complete understanding of how to maximize efficacy of ABL kinase-inhibitor-based therapy, it is important to determine the molecular mechanism and kinetics by which these inhibitors commit CML cells to undergo apoptosis. The surprising clinical observation that dasatinib is effective in vivo despite only transient inhibition of BCR-ABL kinase activity challenges the dogma that kinase inhibitor efficacy is predicated on continual target inhibition. A detailed investigation of the core requirements for an ABL kinase inhibitor to irrevocably commit BCR-ABL-positive cells to apoptosis in CML will shed light on this process. Finally, further insight into the regulation of ABL kinase activity in the context of the multimeric oncogenic kinase BCR-ABL will be critical to improvements in our understanding of the biology and treatment of CML. We will apply a systematic approach to generate, screen, and pursue BCR-ABL protein constructs that are candidates for x-ray crystallography. Structural studies would enhance our understanding of the structural basis for activation and regulation of this oncogenic kinase. Thus, this proposal centers on: 1) elucidation of resistance mechanisms in CML, 2) identification of critical apoptosis commitment pathways associated with tyrosine kinase inhibitor sensitivity, and 3) improving our understanding of the regulation of kinase activity of BCR-ABL. This knowledge will inform and improve the disease management for patients with CML and other malignancies.

Public Health Relevance

Targeted therapy for chronic myeloid leukemia (CML) with imatinib (Gleevec) has dramatically altered the prognosis for patients with this disease, but resistance to imatinib occurs in some patients. The proposed studies will improve our understanding and ability to control resistance, aid in the design of novel therapies by better understanding how cells are killed by imatinib, and provide insight into how the target of imatinib causes the growth of leukemia cells. Together, these findings will have important implications for the treatment of CML and for the use of targeted therapies in many other cancers.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37CA065823-17
Application #
8245031
Study Section
Cancer Molecular Pathobiology Study Section (CAMP)
Program Officer
Mufson, R Allan
Project Start
1995-07-01
Project End
2016-02-29
Budget Start
2012-03-01
Budget End
2013-02-28
Support Year
17
Fiscal Year
2012
Total Cost
$310,293
Indirect Cost
$108,804
Name
Oregon Health and Science University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
Watanabe-Smith, Kevin; Godil, Jamila; Agarwal, Anupriya et al. (2017) Analysis of acquired mutations in transgenes arising in Ba/F3 transformation assays: findings and recommendations. Oncotarget 8:12596-12606
Pietarinen, Paavo O; Eide, Christopher A; Ayuda-DurĂ¡n, Pilar et al. (2017) Differentiation status of primary chronic myeloid leukemia cells affects sensitivity to BCR-ABL1 inhibitors. Oncotarget 8:22606-22615
Watanabe-Smith, K; Tognon, C; Tyner, J W et al. (2016) Discovery and functional characterization of a germline, CSF2RB-activating mutation in leukemia. Leukemia 30:1950-3
Zabriskie, M S; Eide, C A; Yan, D et al. (2016) Extreme mutational selectivity of axitinib limits its potential use as a targeted therapeutic for BCR-ABL1-positive leukemia. Leukemia 30:1418-21
Jacob, Thomas; Agarwal, Anupriya; Ramunno-Johnson, Damien et al. (2016) Ultrasensitive proteomic quantitation of cellular signaling by digitized nanoparticle-protein counting. Sci Rep 6:28163
Davare, Monika A; Vellore, Nadeem A; Wagner, Jacob P et al. (2015) Structural insight into selectivity and resistance profiles of ROS1 tyrosine kinase inhibitors. Proc Natl Acad Sci U S A 112:E5381-90
Beer, Philip A; Knapp, David J H F; Miller, Paul H et al. (2015) Disruption of IKAROS activity in primitive chronic-phase CML cells mimics myeloid disease progression. Blood 125:504-15
Deininger, Michael W; Kopecky, Kenneth J; Radich, Jerald P et al. (2014) Imatinib 800 mg daily induces deeper molecular responses than imatinib 400 mg daily: results of SWOG S0325, an intergroup randomized PHASE II trial in newly diagnosed chronic phase chronic myeloid leukaemia. Br J Haematol 164:223-32
Agarwal, Anupriya; Mackenzie, Ryan J; Besson, Arnaud et al. (2014) BCR-ABL1 promotes leukemia by converting p27 into a cytoplasmic oncoprotein. Blood 124:3260-73
Agarwal, Anupriya; MacKenzie, Ryan J; Pippa, Raffaella et al. (2014) Antagonism of SET using OP449 enhances the efficacy of tyrosine kinase inhibitors and overcomes drug resistance in myeloid leukemia. Clin Cancer Res 20:2092-103

Showing the most recent 10 out of 39 publications