Combination of Bcl2 Antagonist with Pim Kinase Inhibitor

Combination of Bcl-2 Antagonist with Pim Kinase Inhibitor in Chronic Lymphocytic Leukemia Bethany La. Vergne and Curt Phifer, Ph. D. at Northwestern State University Fabiola Gomez, Ph. D. and Varsha Gandhi, Ph. D. ; MD Anderson Cancer Center

What is cancer? Why is it important? �“A disease in which abnormal cells divide without control and can invade nearby tissues” (NCI dictionary) �Enhanced survival mechanisms difficult to kill cancer cells �Physical, emotional, and financial impact �Prevention and treatment protocols for many cancers are available

Chronic Lymphocytic Leukemia (CLL) �Leukemia = cancer of the blood �Lymphocytic = B-lymphocytes �Chronic = slow, gradual development �Patients are 60 -80 years old; rarely under 40 �Complications with immune system �Family history is good predictor of risk �Elevated levels of Bcl-2 and Pim kinase

Apoptosis in lymphocytes

Bcl-2 antagonist and Pim kinase inhibitor �ABT-737 (Abbot Labs) binds to and sequesters Bcl-2 �AZD-1208 (Astra Zeneca) inhibits Pim kinase function

Apoptosis in CLL

Method �Blood samples were obtained from 7 CLL patients and 1 healthy blood donor �Mononuclear cells (including B-lymphocytes) were isolated from blood �Each sample was treated 24 hours total �Cell death determined by Annexin V/Propidium iodide staining �Additive, Antagonistic, and Synergistic

ABT-737 Dose Response 1 nm was used in combination treatments 3μM and 10μM AZD-1208 were determined sufficient by a similar experiment

Combination Treatment Results 1 n. M ABT-737 3μM AZD-1208 10μM AZD-1208 1 n. M ABT-737 + 3μM AZD-1208 1 n. M ABT-737 + 10μM AZD-1208

Combination Treatment Results 1 n. M ABT-737 3μM AZD-1208 10μM AZD-1208 1 n. M ABT-737 + 3μM AZD-1208 1 n. M ABT-737 + 10μM AZD-1208 No treatment DMSO

Conclusions �Variation between samples �Additive results at best �Highest % cell death induced: 40% from 1 n. M ABT-737 + 10μM AZD-1208

References Kang, M, Reynolds, C. “Bcl-2 Inhibitors: Targeting Mitochondrial Apoptotic Pathways in Cancer Therapy. ” Clinical Cancer Research 2009, 15: 1126 -1132. Martinou, J, Youle, R. “Mitochondria in Apoptosis: Bcl-2 Family Members and Mitochondrial Dynamics. ” Developmental Cell 2011, 21: 92 -101. Nawijn, M, Alendar, A. Berns, A. For better or for worse: the role of Pim oncogenes in tumorigenesis. Nature Reviews 2011, 72: 23 -34. Perez, K, Winer, E. “Chronic Lymphocytic Leukemia: Something Old, Something New and Something Borrowed…” Rhode Island Medicine and Health 2011, 94: 1518. Reed, J, Pellecchia, M. Apoptosis-based therapies for hematologic malignancies. Blood 2005, 106: 408 -416. Song, J, Kraft, A. Pim kinase inhibitors sensitize prostate cancer cells to apoptosis triggered by Bcl-2 family inhibitor ABT-737. Cancer Research 2012, 72: 294 -303.