Abstract A32: Adipose-derived mesenchymal stem cell has the differentiation/reprogramming capacity towards two distinct cancer-associated fibroblasts

Cancer-associated fibroblasts (CAFs) are well known of its heterogeneity with distinct phenotypes and functions in tumor progression. CAFs are believed to be composed mainly of resident tissue fibroblast, such as pancreas stellate cells, but bone marrow-derived mesenchymal stem cells (MSC) are also known to be their source (Ishii G et al., Adv Drug Deliv Rev 2016). Herein, we focus on adipose-derived MSC (AD-MSC) as an alternative source of CAF, as one has high reactivity of differentiation and reprogramming to the many cell types (Ronnov-Jessen L et al., J Clin Invest 1995; Sugii/Kida et al., PNAS 2010). It was reported that CAFs include myofibroblastic CAFs (myCAFs), which are found adjacent to tumor cells, and inflammatory CAFs (iCAFs), which localize farther away within the dense stroma (Ohlund, Tuveson et al., J Exp Med 2017). We investigated whether AD-MSC can be a CAF progenitor and differentiated to the different subpopulations of CAFs by employing different culture condition. To evaluate the differentiation capacity, a GFP labeled AD-MSC cell line and the human pancreatic-cancer cell line (Capan-1) were co-cultured in two condition, including direct (DR) co-culture or indirect transwell (TW) co-culture. We then performed qRT-PCR analysis and immunocytochemistry to examine their myCAF marker (ACTA-2, CTGF) or iCAF markers (IL-6, CXCL1, LIF) and compared with those in monocultured AD-MSC. We also analyzed in vitro tumor formation in DR co-culture using time-lapse imaging. In DR co-culture, the morphology of Capan-1 and AD-MSC recapitulated the clinical pancreatic cancer, with those CAF markers expressions. In the course of co-culture, sequential morphologic alterations were observed in AD-MSCs. The AD-MSC with stellate shape changed to large and small spindle-like cells through crosstalk with cancer cells. In sharp contrast, only iCAF marker expression was elevated in TW co-culture. These results collectively suggest that the AD-MSC have a potential to be progenitors of both myCAF and iCAF, explaining the heterogeneity of CAFs under the direct crosstalk with cancer cells. In the presentation, we discuss the differentiation/reprogramming capacity of AD-MSC towards two distinct CAFs (i.e., myCAF and iCAF), focusing on the cell morphology and RNA-seq analysis. Citation Format: Yoshihiro Miyazaki, Yutato Kumagai, Hiroko Kushige, Osamu Shimomura, Yasuyuki Kida, Tatsuya Oda. Adipose-derived mesenchymal stem cell has the differentiation/reprogramming capacity towards two distinct cancer-associated fibroblasts [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2019 Sept 6-9; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2019;79(24 Suppl):Abstract nr A32.