Postnatal Abrogation of VEGFR2 Blocks Terminal Cap2 Differentiation by Preventing the Developmental Progression from...

After birth, the alveolar capillary network expands to increase gas exchange surface area and endothelial-derived signals promote alveolarization. Lung capillaries are comprised of two distinct subsets, one with proliferative potential to facilitate growth and repair (Cap1), and the other serving a specialized role in gas exchange (Cap2). However, the molecular mechanisms directing capillary speciation, developmental plasticity, and fate transitions during development and repair are not well understood. Here, we show that Cap2 are absent in late embryonic life but rapidly appear and expand immediately after birth. We show that Cap1 progenitors first transition to a novel, intermediate cell state (Cap^INT), characterized by co-expression of Cap1 and Cap2 markers, and heightened proliferation. Cap^INT are present in both the developing mouse and human lung. Hyperoxia, an experimental model of bronchopulmonary dysplasia (BPD), a chronic lung disease marked by impaired alveolarization, increases Cap^INT abundance and persistence and expands Cap2 EC. Cap^INT EC are also increased in human infants dying with active BPD. Using genetic lineage tracing, single cell transcriptomics, ATAC-sequencing and a mouse model that permits inducible deletion of VEGFR2 in Cap^INT and Cap2 EC, we show that postnatal abrogation of VEGFR2 markedly increases Cap^INT EC abundance, blocks Cap2 terminal differentiation, impairs alveolarization, and activates alveolar fibroblasts. Finally, we identify ERG as a putative VEGFR2-downstream mechanism that promotes Cap^INT to Cap2 differentiation. Taken together, our data show that Cap1-Cap2 differentiation is a two-step process that only requires VEGFR2 for the second step. Elucidation of the physiologic and molecular pathways that control the initial transition of Cap1 to Cap^INT EC has the potential to reveal new therapeutic targets for lung diseases that disrupt the alveolar capillary formation and integrity.