TGF-ß1-dependent expression of FOXS1 attenuates adipogenic potential and enhances a myofibroblast cellular phenotype [10T_RNAseq]

White adipose tissue (WAT) fibrosis is a major determinant of obesity-induced cardiometabolic dysfunction and is characterized by excessive extracellular matrix (ECM) deposition and myofibroblast activation. Transforming growth factor (TGF)-ß1 is a profibrotic cytokine that potently induces myofibroblast activation in adipocyte stem cells (ASC). How TGF-ß1 orchestrates ASC activation in WAT fibrosis is not completely understood. We identified FOXS1, a member of the forkhead box transcription factor superfamily, as a transcriptional target of TGF-ß1 signaling in primary human WAT ASC (hASC). FOXS1 potentiated TGF-ß1-dependent upregulation of several myofibroblast genes (e.g. Acta2, Col1a1, Fn1, Il11) in 10T1/2 fibroblasts. FOXS1 also mitigated the induction of several adipogenic factors (e.g. Pparg, Stat5a, Fabp4, Adipoq) in 10T1/2 fibroblasts and sensitized these cells to the anti-adipogenic effects of TGF-ß1. Furthermore, loss of endogenous FOXS1 improved adipogenic permissiveness and activated proadipogenic gene programs in 10T1/2 cells, even after TGF-ß1 stimulation. These results indicate that FOXS1 is a positive regulator of profibrotic TGF-ß1-dependent cellular responses, orchestrating the regulation of molecular phenotypes that promote myofibroblast activation and block adipogenesis. These findings offer novel insight into the TGF-ß1-dependent roles of FOXS1 in fibroblasts within the context of profibrotic ASC activation and provide a foundation for further investigation into the role of FOXS1 in WAT fibrosis and obesity-induced cardiometabolic dysfunction. Overall design: RNA-seq profiling of wildtype (WT) or FOXS1 knockout (KO) C3H10T1/2 cells stimulated with TGF-ß1 (1 nM) for 12 hours then stimulated with adipogenic differentiation media (DMII) for 48 hours.