Characterizing Extracellular Matrix Remodeling of Reprogrammed Cancer-Associated Fibroblasts — 105p — Makarie Wiebersick ¹, Karla Arcelay Garcia ¹, Hailey Axemaker ¹, Pilar de la Puente ^1,2,3

¹   Cancer Biology and Immunotherapies Group, Sanford Research, Sioux Falls, SD 57104, USA

²   Department of Obstetrics and Gynecology, University of South Dakota Sanford School of Medicine, Sioux Falls, SD 57105, USA

³   Department of Surgery, University of South Dakota Sanford School of Medicine, Sioux Falls, SD 57105, USA

Ovarian carcinoma (OC) is one of the leading causes of cancer-associated death in women. The five-year survival rate for OC patients is significantly lower than other cancers, at 50%. One of the main contributors to OC progression is extracellular matrix (ECM) remodeling in the tumor microenvironment (TME). In the OC-TME normal fibroblasts are reprogrammed into cancer-associated fibroblasts (CAFs), which are known to promote drug resistance, tumor progression, and metastasis in patients. In this study we produced cell derived extracellular matrices (CD-ECMs) using normal human uterine fibroblasts (HUFs) and normal ovarian fibroblasts (NOFs) as well as reprogrammed CAFs from two different cell lines (KURAMOCHI and SKOV-3) to characterize the ECM remodeling. We plated the different cell types and supplemented cell media with ascorbic acid to promote ECM secretion and cultured for 5-7 days. Using the cellularized ECM, we utilized immunofluorescence staining to characterize ECM marker expression. We also denuded the cells from the ECMs, and added ovarian cancer cells to these decellularized ECMs in order to functionally analyze cell attachment, tumor progression, and drug resistance. In immunofluorescence staining, collagen, laminin, and fibronectin expression were increased in both types of CAFs compared to normal fibroblasts. Cell attachment showed that cancer cells adhere to the CD-ECMs in CAFs at a higher rate compared to normal fibroblasts. Tumor progression and drug resistance was enhanced in the CD-ECMs of uterine and ovarian derived CAFs compared to the normal fibroblasts. In conclusion, our results suggest that the ECM remodeling of CAFs is increasingly dysregulated in comparison to normal fibroblasts. Future studies using these resources could help in identifying the underlying mechanisms of ECM remodeling, which could improve chemotherapy treatments, and improve patient outcomes.

Sanford Research
Pilar de la Puente