{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T05:23:24Z","timestamp":1784697804517,"version":"3.55.0"},"posted":{"date-parts":[[2025,7,28]]},"group-title":"Developmental Biology","reference-count":46,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,7,28]],"date-time":"2025-07-28T00:00:00Z","timestamp":1753660800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nd\/4.0\/"}],"funder":[{"name":"Marc and Lynne Benioff Endowed Chair"},{"name":"NIH NHLBI","award":["R01-HL154002"],"award-info":[{"award-number":["R01-HL154002"]}]},{"name":"NIH NHLBI","award":["HL155828"],"award-info":[{"award-number":["HL155828"]}]},{"name":"NIH NHLBI","award":["HL160018"],"award-info":[{"award-number":["HL160018"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,11,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  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\n                  <jats:sup>INT<\/jats:sup>\n                  ), characterized by co-expression of Cap1 and Cap2 markers, and heightened proliferation. Cap\n                  <jats:sup>INT<\/jats:sup>\n                  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\n                  <jats:sup>INT<\/jats:sup>\n                  abundance and persistence and expands Cap2 EC. Cap\n                  <jats:sup>INT<\/jats:sup>\n                  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\n                  <jats:sup>INT<\/jats:sup>\n                  and Cap2 EC, we show that postnatal abrogation of VEGFR2 markedly increases Cap\n                  <jats:sup>INT<\/jats:sup>\n                  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\n                  <jats:sup>INT<\/jats:sup>\n                  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\n                  <jats:sup>INT<\/jats:sup>\n                  EC has the potential to reveal new therapeutic targets for lung diseases that disrupt the alveolar capillary formation and integrity.\n                <\/jats:p>","DOI":"10.1101\/2025.07.23.666389","type":"posted-content","created":{"date-parts":[[2025,7,29]],"date-time":"2025-07-29T06:45:30Z","timestamp":1753771530000},"source":"Crossref","is-referenced-by-count":5,"title":["Postnatal Abrogation of VEGFR2 Blocks Terminal Cap2 Differentiation by Preventing the Developmental Progression from a Capillary Intermediate Cell State"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7694-8119","authenticated-orcid":false,"given":"Daoqin","family":"Zhang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8691-6428","authenticated-orcid":false,"given":"Carsten","family":"Knutsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David J.","family":"Stroud","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xibing","family":"Che","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1684-2056","authenticated-orcid":false,"given":"David N.","family":"Cornfield","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7097-8539","authenticated-orcid":false,"given":"Fabio","family":"Zanini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6921-0001","authenticated-orcid":false,"given":"Cristina M.","family":"Alvira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025111205500527000_2025.07.23.666389v2.1","doi-asserted-by":"publisher","DOI":"10.1164\/ajrccm.164.10.2101140"},{"key":"2025111205500527000_2025.07.23.666389v2.2","doi-asserted-by":"publisher","DOI":"10.1152\/ajplung.2000.279.3.L600"},{"key":"2025111205500527000_2025.07.23.666389v2.3","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.105.541524"},{"key":"2025111205500527000_2025.07.23.666389v2.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.imbio.2011.03.006"},{"key":"2025111205500527000_2025.07.23.666389v2.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.trsl.2013.11.010"},{"key":"2025111205500527000_2025.07.23.666389v2.6","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.jss.2008.10.007","article-title":"Vascular Endothelial Growth Factor Isoform and Receptor Expression During Compensatory Lung Growth","volume":"160","year":"2010","journal-title":"J. Surg. Res"},{"key":"2025111205500527000_2025.07.23.666389v2.7","doi-asserted-by":"publisher","DOI":"10.1038\/nature17040"},{"key":"2025111205500527000_2025.07.23.666389v2.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2011.10.003"},{"key":"2025111205500527000_2025.07.23.666389v2.9","doi-asserted-by":"publisher","DOI":"10.1038\/nature09493"},{"key":"2025111205500527000_2025.07.23.666389v2.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2009.01.006"},{"key":"2025111205500527000_2025.07.23.666389v2.11","doi-asserted-by":"publisher","DOI":"10.1038\/nature12681"},{"key":"2025111205500527000_2025.07.23.666389v2.12","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.119.313901"},{"key":"2025111205500527000_2025.07.23.666389v2.13","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2822-7"},{"key":"2025111205500527000_2025.07.23.666389v2.14","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2020.01.009"},{"key":"2025111205500527000_2025.07.23.666389v2.15","doi-asserted-by":"crossref","unstructured":"Zanini, F. et al. Developmental diversity and unique sensitivity to injury of lung endothelial subtypes during postnatal growth. iScience 26, 106097 (2023).","DOI":"10.1016\/j.isci.2023.106097"},{"key":"2025111205500527000_2025.07.23.666389v2.16","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-21865-2"},{"key":"2025111205500527000_2025.07.23.666389v2.17","doi-asserted-by":"publisher","DOI":"10.1242\/dev.199512"},{"key":"2025111205500527000_2025.07.23.666389v2.18","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.53072"},{"key":"2025111205500527000_2025.07.23.666389v2.19","doi-asserted-by":"crossref","unstructured":"Ellis, L. V. , Bywaters, J. D. & Chen, J. Endothelial deletion of p53 generates transitional endothelial cells and improves lung development during neonatal hyperoxia. bioRxiv (2024).","DOI":"10.1101\/2024.05.07.593014"},{"key":"2025111205500527000_2025.07.23.666389v2.20","doi-asserted-by":"crossref","unstructured":"Sveiven, S. N. , Knutsen, C. , Zanini, F. , Cornfield, D. & Alvira, C. Single Cell RNA Sequencing Reveals Gene Expression Continuums Along the Spatial Hierarchy of the Pulmonary Circulation. bioRxiv (2025).","DOI":"10.1101\/2025.05.17.654540"},{"key":"2025111205500527000_2025.07.23.666389v2.21","doi-asserted-by":"crossref","unstructured":"Bhattacharya, S. et al. Single-Cell Transcriptomic Profiling Identifies Molecular Phenotypes of Newborn Human Lung Cells. Genes 15, 298 (2024).","DOI":"10.3390\/genes15030298"},{"key":"2025111205500527000_2025.07.23.666389v2.22","doi-asserted-by":"publisher","DOI":"10.1152\/ajplung.00408.2021"},{"key":"2025111205500527000_2025.07.23.666389v2.23","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2020.06.020"},{"key":"2025111205500527000_2025.07.23.666389v2.24","doi-asserted-by":"crossref","unstructured":"Zou, M. et al. Latent Transforming Growth Factor-\u03b2 Binding Protein-2 Regulates Lung Fibroblast-to-Myofibroblast Differentiation in Pulmonary Fibrosis via NF-\u03baB Signaling. Front. Pharmacol. 12, 788714 (2021).","DOI":"10.3389\/fphar.2021.788714"},{"key":"2025111205500527000_2025.07.23.666389v2.25","doi-asserted-by":"publisher","DOI":"10.1096\/fj.201500173"},{"key":"2025111205500527000_2025.07.23.666389v2.26","doi-asserted-by":"publisher","DOI":"10.1126\/scisignal.abb4282"},{"key":"2025111205500527000_2025.07.23.666389v2.27","doi-asserted-by":"publisher","DOI":"10.1164\/rccm.2105057"},{"key":"2025111205500527000_2025.07.23.666389v2.28","doi-asserted-by":"publisher","DOI":"10.1016\/S0002-9343(02)01562-0"},{"key":"2025111205500527000_2025.07.23.666389v2.29","doi-asserted-by":"crossref","unstructured":"Shah, A. V. et al. The endothelial transcription factor ERG mediates Angiopoietin-1-dependent control of Notch signalling and vascular stability. Nat. Commun. 8, 16002 (2017).","DOI":"10.1038\/ncomms16002"},{"key":"2025111205500527000_2025.07.23.666389v2.30","doi-asserted-by":"publisher","DOI":"10.1038\/nature12930"},{"key":"2025111205500527000_2025.07.23.666389v2.31","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2020.06.020"},{"key":"2025111205500527000_2025.07.23.666389v2.32","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2024.12.002"},{"key":"2025111205500527000_2025.07.23.666389v2.33","unstructured":"Cantu, A. et al. Remarkable sex-specific differences at Single-Cell Resolution in Neonatal Hyperoxic Lung Injury. bioRxiv."},{"key":"2025111205500527000_2025.07.23.666389v2.34","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2009.01.006"},{"key":"2025111205500527000_2025.07.23.666389v2.35","doi-asserted-by":"publisher","DOI":"10.1038\/nn.2467"},{"key":"2025111205500527000_2025.07.23.666389v2.36","doi-asserted-by":"crossref","first-page":"L299","DOI":"10.1152\/ajplung.00034.2023","article-title":"Endothelial-specific loss of IKK\u03b2 disrupts pulmonary endothelial angiogenesis and impairs postnatal lung growth","volume":"325","year":"2023","journal-title":"Am. J. Physiol.-Lung Cell. Mol. Physiol"},{"key":"2025111205500527000_2025.07.23.666389v2.37","doi-asserted-by":"publisher","DOI":"10.7554\/elife.56890"},{"key":"2025111205500527000_2025.07.23.666389v2.38","doi-asserted-by":"publisher","DOI":"10.1165\/rcmb.2013-0014TR"},{"key":"2025111205500527000_2025.07.23.666389v2.39","doi-asserted-by":"crossref","unstructured":"Young, M. D. & Behjati, S . SoupX removes ambient RNA contamination from droplet-based single-cell RNA sequencing data. GigaScience 9, (2020).","DOI":"10.1093\/gigascience\/giaa151"},{"key":"2025111205500527000_2025.07.23.666389v2.40","doi-asserted-by":"crossref","unstructured":"Wolf, F. A. , Angerer, P. & Theis, F. J . SCANPY: large-scale single-cell gene expression data analysis. Genome Biol. 19, (2018).","DOI":"10.1186\/s13059-017-1382-0"},{"key":"2025111205500527000_2025.07.23.666389v2.41","doi-asserted-by":"publisher","DOI":"10.1038\/S41576-023-00586-W"},{"key":"2025111205500527000_2025.07.23.666389v2.42","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/j.cels.2018.11.005","article-title":"Scrublet: Computational Identification of Cell Doublets in Single-Cell Transcriptomic Data","volume":"8","year":"2019","journal-title":"Cell Syst"},{"key":"2025111205500527000_2025.07.23.666389v2.43","doi-asserted-by":"crossref","unstructured":"Traag, V. A. , Waltman, L. & Van Eck, N. J . From Louvain to Leiden: guaranteeing well-connected communities. Sci. Rep. 9, (2019).","DOI":"10.1038\/s41598-019-41695-z"},{"key":"2025111205500527000_2025.07.23.666389v2.44","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0414-6"},{"key":"2025111205500527000_2025.07.23.666389v2.45","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0591-3"},{"key":"2025111205500527000_2025.07.23.666389v2.46","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0439-x"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.07.23.666389","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T09:13:11Z","timestamp":1768468391000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.07.23.666389"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,28]]},"references-count":46,"URL":"https:\/\/doi.org\/10.1101\/2025.07.23.666389","relation":{},"subject":[],"published":{"date-parts":[[2025,7,28]]},"subtype":"preprint"}}