Sox9 and Rbpj differentially regulate endothelial to mesenchymal transition and wound scarring in murine endovascular progenitors

Endothelial to mesenchymal transition (EndMT) is a leading cause of fibrosis and disease, however its mechanism has yet to be elucidated. The endothelium possesses a profound regenerative capacity to adapt and reorganize that is attributed to a population of vessel-resident endovascular progenitors (EVP) governing an endothelial hierarchy. Here, using fate analysis, we show that two transcription factors SOX9 and RBPJ specifically affect the murine EVP numbers and regulate lineage specification. Conditional knock-out of Sox9 from the vasculature ( Sox9 fl/fl /Cdh5-Cre ER RosaYFP ) depletes EVP while enhancing Rbpj expression and canonical Notch signalling. Additionally, skin wound analysis from Sox9 conditional knock-out mice demonstrates a significant reduction in pathological EndMT resulting in reduced scar area. The converse is observed with Rbpj conditionally knocked-out from the murine vasculature ( Rbpj fl/fl /Cdh5-CreER RosaYFP ) or inhibition of Notch signaling in human endothelial colony forming cells, resulting in enhanced Sox9 and EndMT related gene ( Snail, Slug, Twist1, Twist2, TGF-β ) expression. Similarly, increased endothelial hedgehog signaling ( Ptch1 fl/fl /Cdh5-CreER RosaYFP ), that upregulates the expression of Sox9 in cells undergoing pathological EndMT, also results in excess fibrosis. Endothelial cells transitioning to a mesenchymal fate express increased Sox9 , reduced Rbpj and enhanced EndMT. Importantly, using topical administration of siRNA against Sox9 on skin wounds can substantially reduce scar area by blocking pathological EndMT. Overall, here we report distinct fates of EVPs according to the relative expression of Rbpj or Notch signalling and Sox9 , highlighting their potential plasticity and opening exciting avenues for more effective therapies in fibrotic diseases. How endothelial to mesenchymal transition is regulated in endovascular progenitors is unclear. Here, the authors show that blocking Sox9 expression in murine endovascular progenitors regulates this transition on skin wounding, affecting the size of scarring, with changes in Rbpj having the opposite effect.