Distinct fibroblast functional states drive clinical outcomes in ovarian cancer and are regulated by TCF21

Recent studies indicate that cancer-associated fibroblasts (CAFs) are phenotypically and functionally heterogeneous. However, little is known about CAF subtypes and the roles they play in cancer progression. Here we identify and characterize two CAF subtypes that coexist within high grade serous ovarian cancers: Fibroblast activation protein (FAP)-high (FH) CAFs resemble the classical myofibroblast-type CAF, whereas FAP-low (FL) CAFs possesses a preadipocyte-like molecular signature. High-grade serous ovarian cancer single cell suspensions of 12 patients were stained for fluorescence activated cell sorting (FACS) CD31-CD45-EpCAM-CD49e+ (CAFs) cells, and CD31-CD45-EpCAM+CD133+ (epithelial) cells were gated for sorting based on FMO controls. Cells were sorted and RNA was extracted immediately using the RNeasy Plus Micro kit (Qiagen).