Selective inhibition of matrix mechanosensing in stromal cells attenuates cardiovascular fibrosis
Matrix-derived biophysical cues are known to regulate the activation of fibroblasts and their subsequent transdifferentiation into myofibroblasts, but whether modulation of these signals can suppress fibrosis in intact tissues remains unclear, particularly in the cardiovascular system. We have employed single-cell/nucleus RNA sequencing of myofibroblasts differentiated from induced pluripotent stem cell-derived cardiac fibroblasts (iPSC-CFs), as well as mouse transverse aortic constriction (TAC) hearts to investigate whether modulation of matrix mechanosensing, in concert with TGF-beta inhibition, can potentiate the de-activation of fibrosis-promoting cardiac stromal populations in vitro and in vivo. Overall design: In vitro samples (scRNA-seq): after 48h of treatment, aSMA-expressing myofibroblasts derived from iPSC-CFs were were harvested (n = 3 wells pooled into a single library per condition) for droplet-based scRNA-seq using the 10X Genomics platform. In vivo samples (snRNSA-seq): following 4 weeks of TAC and 2 weeks of subsequent drug treatment, apical tissue samples were harvested from male C57BL/6 mice (n = 3 animals pooled into a single sample per experimental group). Joint snRNA- and snATAC-seq libraries were prepared using the 10X scMultiome platform, but only the snRNA-seq (GEX) libraries were used for this study.