Abstract 990: Functional genomic screen of kinases in high-grade serous ovarian cancer metastasis

Abstract High-Grade Serous Ovarian Cancer (HGSC) is a highly metastatic cancer with the majority of patients presenting in advanced stages. Currently there are limited targeted treatment options for patients, especially for women with high metastatic tumor burden. In order to improve patient outcomes, we have aimed to identify and characterize kinases that may regulate the steps of metastasis in HGSC. Kinases in particular serve as actionable targets because we can design inhibitors of the kinases and their ligands. Several kinases, such as tyrosine kinases and aurora kinases, have previously been implicated in ovarian cancer and seem to influence tumor cell metastasis. Our lab performed a functional genomic screen on 719 kinases in a tumor cell attachment and proliferation assay. In this screen, we cultured primary fibroblasts from patient omentum and plated them on top of a collagen and fibronectin matrix, to mimic the tumor microenvironment. We used siRNA transfection reagents, which were plated atop the fibroblasts along with GFP-labeled ovarian cancer cells that were then incubated with the other cell layers for 72 hours. Based on the florescence intensities, we calculated median average deviations (MAD) values normalized to controls to be a measure of ovarian cancer cell attachment and proliferation. This screen was replicated in 3 ovarian cancer cell lines, including OVCAR8, ES2, and A2780, across 9 panels for 2 libraries. Both libraries had 2 distinct siRNAs for each kinase, in total 4 siRNAs per kinase. From this initial screen, we have identified 20 candidate kinases to validate based on having significantly negative MAD values in at least two cells lines in the screen. Candidate kinases have been validated by knock-down in two ovarian cancer cell lines, ES2 and OVCAR8, and assessed on their ability to migrate in a wound-healing assay. The top kinases will be further studied for their ability to regulate the steps of tumor cell metastasis and the mechanisms that allow them to promote metastasis. Our study aims to identify actionable targets that influence these steps of metastasis, so we can develop drugs that target metastatic tumors and serve as maintenance therapies to help prevent further metastasis, mediate chemoresistance, and improve patient outcomes. Citation Format: Emilee Nicole Kotnik, Elena Lomonosova, Daniel Wilke, Gregory Longmore, Katherine C. Fuh. Functional genomic screen of kinases in high-grade serous ovarian cancer metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 990.