A Single-Short Partial Reprogramming of the Endothelial Cells in vivo decreases Blood Pressure and Vascular Contractility Through the Attenuation of Endothelial-to-Mesenchymal Transition in Male Hypertensive Mice

Endothelial cell (EC) reprogramming represents an innovative strategy in regenerative medicine to prevent deleterious effects induced by cardiovascular diseases. Here, we showed for the first time, that a single-short partial reprogramming of ECs, via the overexpression of three master transcription factors Oct-3/4, Sox-2, and Klf-4 (OSK), was able to bring ECs back to a youthful phenotype in hypertensive mice. Accordingly, we effectively generated in vitro ECs with endothelial progenitor cell (EPC)-like features that expressed lower migratory capability and prevented cellular senescence. Furthermore, in vivo OSK treatment of hypertensive mice normalized blood pressure and resistance arteries hypercontractility, via the attenuation of the process of endothelial-to-mesenchymal transition (EndMT) and alleviation of elastin breaks in arteries from hypertensive mice. Human hypertensive ECs treated with OSK recovered their ability to activate endothelial nitric oxide synthase (eNOS) and enhanced nitric oxide (NO) production, with a decrease in reactive oxygen species (ROS) generation. In addition, single-cell RNA analysis showed that OSK could alleviate EC senescence and mitigate the EndMT process, facilitating the restoration of EC phenotypes in human hypertensive ECs. Overall design: Human male and female normal and hypertensive aortic endothelial cells were treated with OSK or control EGFP sendai virus for 3 or 7 days. The attentuation of EndMT in HAoECs by OSK were determined using scRNA-seq.