title: R-Ketorolac Targets Cdc42 and Rac1 and Alters Ovarian Cancer Cell Behaviors Critical for Invasion and Metastasis.
abstract: Cdc42 (cell division control protein 42) and Rac1 (Ras-related C3 botulinum toxin substrate 1) are attractive therapeutic targets in ovarian cancer based on established importance in tumor cell migration, adhesion, and invasion. Despite a predicted benefit, targeting GTPases has not yet been translated to clinical practice. We previously established that Cdc42 and constitutively active Rac1b are overexpressed in primary ovarian tumor tissues. Through high-throughput screening and computational shape homology approaches, we identified R-ketorolac as a Cdc42 and Rac1 inhibitor, distinct from the anti-inflammatory, cyclooxygenase inhibitory activity of S-ketorolac. In the present study, we establish R-ketorolac as an allosteric inhibitor of Cdc42 and Rac1. Cell-based assays validate R-ketorolac activity against Cdc42 and Rac1. Studies on immortalized human ovarian adenocarcinoma cells (SKOV3ip) and primary patient-derived ovarian cancer cells show that R-ketorolac is a robust inhibitor of growth factor or serum-dependent Cdc42 and Rac1 activation with a potency and cellular efficacy similar to small-molecule inhibitors of Cdc42 (CID2950007/ML141) and Rac1 (NSC23766). Furthermore, GTPase inhibition by R-ketorolac reduces downstream p21-activated kinases (PAK1/PAK2) effector activation by >80%. Multiple assays of cell behavior using SKOV3ip and primary patient-derived ovarian cancer cells show that R-ketorolac significantly inhibits cell adhesion, migration, and invasion. In summary, we provide evidence for R-ketorolac as a direct inhibitor of Cdc42 and Rac1 that is capable of modulating downstream GTPase-dependent, physiologic responses, which are critical to tumor metastasis. Our findings demonstrate the selective inhibition of Cdc42 and Rac1 GTPases by an FDA-approved drug, racemic ketorolac, that can be used in humans.
authors.affiliation: Department of Pathology, University of New Mexico School of Medicine, Albuquerque, New Mexico. Cancer Center, University of New Mexico, Albuquerque, New Mexico.
authors.forename: Yuna
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Cancer Center, University of New Mexico, Albuquerque, New Mexico. Department of Pharmaceutical Sciences, University of New Mexico College of Pharmacy, Albuquerque, New Mexico.
authors.forename: S Ray
authors.initials: SR
authors.lastname: Kenney
authors.affiliation: Cancer Center, University of New Mexico, Albuquerque, New Mexico. Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of New Mexico School of Medicine, Albuquerque, New Mexico.
authors.forename: Carolyn Y
authors.initials: CY
authors.lastname: Muller
authors.affiliation: Cancer Center, University of New Mexico, Albuquerque, New Mexico. Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of New Mexico School of Medicine, Albuquerque, New Mexico.
authors.forename: Sarah
authors.initials: S
authors.lastname: Adams
authors.affiliation: Cancer Center, University of New Mexico, Albuquerque, New Mexico. Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of New Mexico School of Medicine, Albuquerque, New Mexico.
authors.forename: Teresa
authors.initials: T
authors.lastname: Rutledge
authors.affiliation: Department of Pathology, University of New Mexico School of Medicine, Albuquerque, New Mexico.
authors.forename: Elsa
authors.initials: E
authors.lastname: Romero
authors.affiliation: Division of Epidemiology, Biostatistics and Preventive Medicine, Department of Internal Medicine, University of New Mexico School of Medicine, Albuquerque, New Mexico.
authors.forename: Cristina
authors.initials: C
authors.lastname: Murray-Krezan
authors.affiliation: Department of Cell and Developmental Biology, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
authors.forename: Rytis
authors.initials: R
authors.lastname: Prekeris
authors.affiliation: Department of Pathology, University of New Mexico School of Medicine, Albuquerque, New Mexico. Cancer Center, University of New Mexico, Albuquerque, New Mexico.
authors.forename: Larry A
authors.initials: LA
authors.lastname: Sklar
authors.affiliation: Cancer Center, University of New Mexico, Albuquerque, New Mexico. Department of Pharmaceutical Sciences, University of New Mexico College of Pharmacy, Albuquerque, New Mexico.
authors.forename: Laurie G
authors.initials: LG
authors.lastname: Hudson
authors.affiliation: Department of Pathology, University of New Mexico School of Medicine, Albuquerque, New Mexico. Cancer Center, University of New Mexico, Albuquerque, New Mexico. wness@unm.edu.
authors.forename: Angela
authors.initials: A
authors.lastname: Wandinger-Ness
mesh_terms: D000494:Allosteric Regulation; D000634:Aminoquinolines / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D000077216:Carcinoma, Ovarian Epithelial; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002465:Cell Movement; D004305:Dose-Response Relationship, Drug; D005260:Female; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D020910:Ketorolac / Q000494:pharmacology*; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D009375:Neoplasms, Glandular and Epithelial / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D011485:Protein Binding; D011554:Pseudopodia; D011743:Pyrimidines / Q000494:pharmacology; D015398:Signal Transduction; D020764:cdc42 GTP-Binding Protein / Q000037:antagonists & inhibitors* / Q000378:metabolism; D020830:rac1 GTP-Binding Protein / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D000634:Aminoquinolines; D000970:Antineoplastic Agents; C487513:NSC 23766; D011743:Pyrimidines; C107953:RAC1 protein, human; D006160:Guanosine Triphosphate; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein; D020910:Ketorolac