Abstract 2571: Effects of patient-derived ovarian cancer-associated fibroblasts and tumor progression and metastasis

Abstract Background: Ovarian cancer (OC) is a highly treatment-resistant malignancy, often diagnosed late due to minimal symptoms and extensive metastasis. Despite platinum-based chemotherapy, recurrence and chemoresistance highlight the need for new therapies. This study focuses on cancer-associated fibroblasts (CAFs), key TME components, to explore their impact on OC progression and identify novel CAF subtypes. Methods: CAFs were isolated from OC patient tumors via enzymatic dissociation and flow cytometry. Their markers, FAP and α-SMA, were analyzed using immunofluorescence and flow cytometry. Conditioned media from CAF cultures were tested on OC cell lines (A2780, A2780-cp, SKOV3, OVCAR3) to assess proliferation and migration via cell viability and scratch assays. Cytokines in CAF-conditioned media were quantified using ELISA. Results: CAFs were identified within OC tumors, with their expression of FAP validated in high-grade serous ovarian cancer (HGSOC) samples via immunohistochemistry and flow cytometry. CAFs were successfully isolated and cultured from patient-derived OC tumors. Flow cytometry revealed that CAFs represent a minor subset of the total tumor cell population. Morphologically, CAFs exhibited plump spindle or stellate forms. CAFs derived from HGSOC displayed elevated levels of FAP and α-SMA expression compared to those derived from non-HGSOC tumors. Exposure of four OC cell lines (A2780, A2780-cp, SKOV3, OVCAR3) to CAF-conditioned media significantly promoted their growth. Scratch assays revealed enhanced motility in two OC cell lines (SKOV3 and OVCAR3) following treatment with CAF-conditioned media. Notably, the HGSOC-derived CAF subtype, W-OVCAF7, exhibited significantly greater growth- and motility-enhancing effects compared to non-HGSOC-derived CAFs. Scratch assays further confirmed that W-OVCAF7 markedly increased the migratory capacity of both SKOV3 and OVCAR3 cells, suggesting that CAF activity may vary depending on the tumor cell type. These findings underscore the pivotal role of CAFs in modulating the proliferation and migration of OC cells. Conclusion: Tumor-derived CAFs significantly enhance OC cell proliferation and migration, underscoring their critical role in the TME. This study highlights the importance of patient-derived CAFs for precise analysis of OC dynamics and lays a foundation for therapeutic strategies targeting the CAF-TME interplay. Citation Format: Ji-Young Lee, Sung Wan Kang, Min-Seo Lee, Ok-Ju Kang, Yong-Man Kim, Shin-Wha Lee. Effects of patient-derived ovarian cancer-associated fibroblasts and tumor progression and metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 2571.