Normalization of vascular intussusception is essential for promoting lung regeneration over fibrosis

Proliferation of vascular endothelial cells (ECs) is critical for providing sufficient cells lining the neo-vascular lumen. Moreover, expanding ECs need to undergo multiple steps to line new lumen, functionalizing anti-thrombotic and anti-inflammatory interface that prevents extravasation of inflammatory cells, maintains blood perfusion, and facilitates cellular crosstalk. Therefore, productive angiogenesis in regeneration requires formation of endothelial tight junction, arrangement of endothelial apical lumen, and alignment of basolateral side juxtaposed with alveolar epithelial cells. As a result, we used single cell RNA sequencing (scRNA-seq) to detect the function of diverse EC clusters. Overall design: 7 days after PNX, wild type mice and their parallel control (Mettl3 KO) were harvested as well as Sham group. The lung tissues were cut off and digested to get cell suspension. Immunomagnetic bead cell sorting was used to isolated mouse lung vascular ECs and magnetic beads were pre-coated with CD31 and CD45 antibody respectively. The endothelial cells were obtained as CD45 negative and CD31 positive cells.