Apoptotic cell death induced by chemotherapeutics and other cell-damaging agents typically involves release of cytochrome c from the mitochondrial intermembrane space to the cytoplasm, where it induces the oligomerization of Apaf-1 to form the """"""""apoptosome"""""""" and activate procaspase 9. Active caspase 9 then activates the executioner caspases 3 and 7 to dismantle the doomed cell. Impaired function or formation of the apoptosome is a hallmark of many cancers, including ovarian cancers, melanoma, lung cancers and others. We have demonstrated that apoptosome formation is defective in leukemias expressing activated tyrosine kinases (Bcr-Abl, Tel-PDGFR2, and activated FLT3). This defect was traced to tight inhibitory binding of the beta isoform of Hsp90 to Apaf-1 in leukemic, but not normal cells. Specifically, a difference in the phosphorylation status of Hsp902 at two sites (S226/S255) could account for the difference in Apaf-1 binding and cytochrome c-induced caspase activation between leukemic and untransformed cells. Moreover, expression of a non- phosphorylatable Hsp902 variant that mimicked the leukemic form conferred resistance to cytochrome c-induced apoptosis in untransformed cells and promoted resistance to the front-line therapeutic imatinib in Bcr-Abl-transformed bone marrow-derived hematopoietic stem cells in vitro. These findings point to the kinases/phosphatases regulating Hsp90 as potentially attractive therapeutic targets.
The aims of this proposal are to elucidate how Hsp902 phosphorylation is misregulated in leukemic cells, to determine how Hsp902 hypophosphorylation impacts Apaf-1 and other Hsp902 targets, to analyze the effects of Hsp902 hypophosphorylation in animals and to evaluate Hsp902 phosphorylation status in patient samples. The long term goal of this work is to fully understand the molecular basis for apoptosome resistance in leukemia and, ultimately, to translate this knowledge into therapeutic agents for leukemias.

Public Health Relevance

This proposal addresses the molecular pathways linking activated tyrosine kinases to the suppression of programmed cell death (apoptosis) in leukemias. Understanding these pathways may have broad implications for the treatment of leukemias and other cancers expressing activated tyrosine kinases. The goal of this work is to fully understand these signaling pathways in order to design better therapeutic agents for the treatment of leukemias.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA102707-09
Application #
8433980
Study Section
Cancer Molecular Pathobiology Study Section (CAMP)
Program Officer
Salnikow, Konstantin
Project Start
2003-07-01
Project End
2015-01-31
Budget Start
2013-02-01
Budget End
2014-01-31
Support Year
9
Fiscal Year
2013
Total Cost
$247,272
Indirect Cost
$88,764
Name
Duke University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
Matsuura, K; Huang, N-J; Cocce, K et al. (2017) Downregulation of the proapoptotic protein MOAP-1 by the UBR5 ubiquitin ligase and its role in ovarian cancer resistance to cisplatin. Oncogene 36:1698-1706
Yang, C-S; Sinenko, S A; Thomenius, M J et al. (2014) The deubiquitinating enzyme DUBAI stabilizes DIAP1 to suppress Drosophila apoptosis. Cell Death Differ 21:604-11
Kurokawa, Manabu; Ito, Takahiro; Yang, Chih-Sheng et al. (2013) Engineering a BCR-ABL-activated caspase for the selective elimination of leukemic cells. Proc Natl Acad Sci U S A 110:2300-5
Parrish, Amanda B; Freel, Christopher D; Kornbluth, Sally (2013) Cellular mechanisms controlling caspase activation and function. Cold Spring Harb Perspect Biol 5:
Kurokawa, Manabu; Kim, Jiyeon; Geradts, Joseph et al. (2013) A network of substrates of the E3 ubiquitin ligases MDM2 and HUWE1 control apoptosis independently of p53. Sci Signal 6:ra32
Chen, Chen; Zhang, Liguo; Huang, Nai-Jia et al. (2013) Suppression of DNA-damage checkpoint signaling by Rsk-mediated phosphorylation of Mre11. Proc Natl Acad Sci U S A 110:20605-10
Kim, Jiyeon; Parrish, Amanda B; Kurokawa, Manabu et al. (2012) Rsk-mediated phosphorylation and 14-3-3? binding of Apaf-1 suppresses cytochrome c-induced apoptosis. EMBO J 31:1279-92
Zhang, Liguo; Huang, Nai-Jia; Chen, Chen et al. (2012) Ubiquitylation of p53 by the APC/C inhibitor Trim39. Proc Natl Acad Sci U S A 109:20931-6
Huang, Nai-Jia; Zhang, Liguo; Tang, Wanli et al. (2012) The Trim39 ubiquitin ligase inhibits APC/CCdh1-mediated degradation of the Bax activator MOAP-1. J Cell Biol 197:361-7
Johnson, C E; Freel, C D; Kornbluth, S (2010) Features of programmed cell death in intact Xenopus oocytes and early embryos revealed by near-infrared fluorescence and real-time monitoring. Cell Death Differ 17:170-9

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