Abstract B81: Selected NSAIDs target GTPases for ovarian cancer therapy
Authors: Guo Yuna , Kenney S. Ray , Romero Elsa , Oprea Tudor , Adams Sarah , Muller Carolyn , Sklar Larry , Buranda Tione , Hudson Laurie , Wandinger-Ness Angela 
Published: August 2014

Abstract
Rac1 and Cdc42 GTPases are key regulators of actin reorganization, cell mobility, cell-cell and cell-extracellular matrix (ECM) adhesion, whose activities are found closely correlated with tumor cell properties (increased proliferation, migration and adhesion), which are important for tumor expansion and malignant progression. We found that both the constitutively active Rac1b and Cdc42 GTPases are overexpressed in primary ovarian tumors and cancer cell lines. Using high throughput screening and computational methods, selected non-steroidal anti-inflammatory drugs (NSAIDs) were identified as regulators of Rac1 and Cdc42 in vitro, which we confirmed in cellular assays and xenograft animal model studies. Among the NSAIDs, ketorolac, inhibited cell adhesion, migration and invasion. When administrated intraperitoneally (IP) in a human xenograft model of ovarian cancer a 3-4 fold reduction in tumor cell number and decreased total tumor burden resulted. The S-enantiomer of ketorolac is a well-established inhibitor of cyclooxygenase (COX) enzymes. We determined that R-enantiomer had little activity against COX and using G-LISA assays to measure GTPase activation established that R-, but not S-ketorolac was an effective GTPase inhibitor. These data prompted us to conduct a clinical pilot study in ovarian cancer patients assessing the bioavailability and impact of clinically approved Toradol (racemic [R, S] ketorolac administrated clinically for pain after surgery) on tumor cell behaviors. Patient serum and ascites samples analyzed by HPLC established that ketorolac was detectable in the ascites fluid within 1 hour after intravenous (IV) administration with predominantly the R-form persisting after 6 hours. Serum concentrations were found to predict ascites concentrations. Purified tumor cells from fresh patient ascites (identified by tumor markers MUC16, EpCAM) when treated in vitro with ketorolac exhibited reduced adhesion to ECM; furthermore, GTPase activity assays demonstrated that R-, but not S-ketorolac significantly inhibited Cdc42 activity. Post-Toradol treatment patient ascites samples also exhibited reduced Cdc42 activity at all timepoints. In sum, we show that the R-enantiomers of selected NSAIDs are novel chemical entities that selectively target Rho-family GTPases and facilitate the development of new chemical entities based on these scaffolds. Such mechanistic tumor-relevant functional activities have not been described previously and suggest that investigation of FDA approved NSAIDs offer opportunities for repurposing and rapid human translation for the treatment of ovarian cancer. The composite results validate Rho-family GTPases and their downstream effectors as potential prognostic indicators and as therapeutic targets for ovarian cancer.
Citation Format: Yuna Guo, S. Ray Kenney, Elsa Romero, Tudor Oprea, Sarah Adams, Carolyn Muller, Larry Sklar, Tione Buranda, Laurie Hudson, Angela Wandinger-Ness. Selected NSAIDs target GTPases for ovarian cancer therapy. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B81.