Although most ovarian cancer patients respond very well to first-line clinical therapy, the vast majority eventually succumb to recurrent, chemoresistant disease. This highlights the urgent need for studies that investigate mechanisms of relapse and identify potential therapeutic targets for the treatment of recurrent ovarian cancer Our lab previously identified ovarian cancer stem cells (OvCSC) marked by coexpression of ALDH and CD133, and discovered that these cells preferentially express high Nuclear Factor of Activated T-Cells 3 (NFAT3) levels. We have observed that cells with a constitutively active NFAT3 (cNFAT3) demonstrate dramatic growth reduction in vitro and significantly smaller tumor size in vivo. Furthermore, cNFAT3 cells are more resistant to cisplatin therapy than control cells; conversely, treatment with VIVIT, an NFAT inhibitor, sensitizes ovarian cancer cells to cisplatin. We hypothesize that NFAT3-induced cell cycle retardation decreases the sensitivity of ovarian cancer stem cells to chemotherapy, thereby allowing them to survive through treatment and later cause recurrent disease. We therefore propose to investigate mechanisms of quiescence and chemoresistance in OvCSC, with the ultimate aim of elucidating the role of OvCSC in relapse. Cell cycle analysis and quiescence-specific FACS analysis will be used to clarify the role of NFAT3 in growth retardation. Array analysis will be used to identify NFAT3-regulated cell cycle targets for further analysis and identification of potential therapeutic targets. We will usea novel single-cell microfluidics culture device that allows direct quantification of asymmetric vs. symmetric divisions to study the effects of NFAT3 on stem cell fate decisions. We will subsequently investigate the effects that NFAT3 has on chemoresistance and tumorigenesis in vitro and in vivo. We will also investigate quiescence-associated proteins and classic chemoresistance mechanisms, such as expression of P-glycoprotein, to identify the mechanism by which NFAT3 protects cells during cisplatin therapy. Finally, primary ovarian tumor samples will be studied to correlate NFAT3 levels with progression free survival, overall survival, and the presence of chemoresistant disease. Taken together, these studies of quiescence and chemoresistance will provide critical insight into mechanisms by which ovarian cancer cells and CSC are able to survive through cisplatin therapy, causing relapse.

Public Health Relevance

Due to a high relapse rate despite initially successful chemotherapy, ovarian cancer is the fifth leading cause of cancer deaths in American women. This proposal aims to investigate quiescence as a mechanism that allows ovarian cancer stem cells to survive through chemotherapy and repopulate tumors. Investigating chemotherapy resistance in these cancer stem cells may provide new therapeutic targets for the prevention or treatment of relapsed and recurrent ovarian cancer.

Agency
National Institute of Health (NIH)
Type
Individual Predoctoral NRSA for M.D./Ph.D. Fellowships (ADAMHA) (F30)
Project #
1F30CA189316-01
Application #
8780177
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Damico, Mark W
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109