Ovarian cancer patients are burdened with a lack of effective therapeutic options resulting in a low survival rate due to chemotherapy resistance, recurrence, and metastasis. The overall goal of the proposed project is to establish innovative anti-cancer drug targets by mapping mechanical regulators of ovarian cancer progression. Cancer metastasis requires cells to deform and migrate through confined spaces. To survive, cancer cells must sense physical forces and adapt to maintain cellular and nuclear mechanical homeostasis. If we could map the mechanical regulators of cancer cells—molecules that control cell deformability and migration—this would enable us to define novel, complementary treatment strategies. To identify novel mechanical regulators, we developed a high throughput deformability screen that tested the effects of 1280 compounds (Library of Pharmacologically Active Compounds) on the deformability of human high-grade serous ovarian cancer (OVCAR5) cells. Our screen revealed drug compounds that reduced cell deformability. A meta-analysis across top drug hits revealed NUDT5 as a predicted regulator of cellular mechanical behaviors. NUDT5 is a member of the Nudix (nucleoside diphosphate linked moiety X) hydrolase superfamily. Our preliminary analyses revealed NUDT5 is highly expressed in various cancer types compared to normal tissue. My preliminary data shows that NUDT5 regulates cell stiffness, morphology, and migration in OVCAR5 cells. The goal of my project is to test the hypothesis that NUDT5 drives ovarian cancer progression by regulating cancer cell mechanical behaviors. In Aim 1, I will assess how NUDT5 relates to ovarian cancer progression by measuring NUDT5 expression and localization at different disease stages in distinct histological subtypes, including high-grade serous (HGSOC) and clear cell (OCCC) carcinoma tumors using patient tissue microarrays. I will also examine in vitro how NUDT5 regulates PARP inhibitor resistance and ovarian cancer cell behaviors, such as proliferation, migration, spheroid formation, and invasion. In Aim 2, I will determine the role of NUDT5 activity in catalyzing ATP production for actin cytoskeletal and nuclear remodeling following confined migration. Understanding how NUDT5 regulates cancer cell mechanical behaviors will guide future clinical treatment strategies to improve patient survival.