{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T02:49:58Z","timestamp":1787021398766,"version":"build-2736575974"},"reference-count":53,"publisher":"The Company of Biologists","issue":"11","license":[{"start":{"date-parts":[[2024,5,31]],"date-time":"2024-05-31T00:00:00Z","timestamp":1717113600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/www.biologists.com\/user-licence-1-1\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["5R01EY031593"],"award-info":[{"award-number":["5R01EY031593"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,6,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Prolyl hydroxylase domain (PHD) proteins are oxygen sensors that use intracellular oxygen as a substrate to hydroxylate hypoxia-inducible factor (HIF) \u03b1 proteins, routing them for polyubiquitylation and proteasomal degradation. Typically, HIF\u03b1 accumulation in hypoxic or PHD-deficient tissues leads to upregulated angiogenesis. Here, we report unexpected retinal phenotypes associated with endothelial cell (EC)-specific gene targeting of Phd2 (Egln1) and Hif2alpha (Epas1). EC-specific Phd2 disruption suppressed retinal angiogenesis, despite HIF\u03b1 accumulation and VEGFA upregulation. Suppressed retinal angiogenesis was observed both in development and in the oxygen-induced retinopathy (OIR) model. On the other hand, EC-specific deletion of Hif1alpha (Hif1a), Hif2alpha, or both did not affect retinal vascular morphogenesis. Strikingly, retinal angiogenesis appeared normal in mice double-deficient for endothelial PHD2 and HIF2\u03b1. In PHD2-deficient retinal vasculature, delta-like 4 (DLL4, a NOTCH ligand) and HEY2 (a NOTCH target) were upregulated by HIF2\u03b1-dependent mechanisms. Inhibition of NOTCH signaling by a chemical inhibitor or DLL4 antibody partially rescued retinal angiogenesis. Taken together, our data demonstrate that HIF2\u03b1 accumulation in retinal ECs inhibits rather than stimulates retinal angiogenesis, in part by upregulating DLL4 expression and NOTCH signaling.<\/jats:p>","DOI":"10.1242\/dev.202802","type":"journal-article","created":{"date-parts":[[2024,5,21]],"date-time":"2024-05-21T07:42:22Z","timestamp":1716277342000},"source":"Crossref","is-referenced-by-count":7,"title":["Endothelial HIF2\u03b1 suppresses retinal angiogenesis in neonatal mice by upregulating NOTCH signaling"],"prefix":"10.1242","volume":"151","author":[{"given":"Li-Juan","family":"Duan","sequence":"first","affiliation":[{"name":"Center for Vascular Biology, University of Connecticut Health Center 1 , Farmington, CT 06030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3835-2944","authenticated-orcid":false,"given":"Yida","family":"Jiang","sequence":"additional","affiliation":[{"name":"Center for Vascular Biology, University of Connecticut Health Center 1 , Farmington, CT 06030, USA"},{"name":"University of Connecticut Health Center 2 Department of Cell Biology , , Farmington, CT 06030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5792-6486","authenticated-orcid":false,"given":"Guo-Hua","family":"Fong","sequence":"additional","affiliation":[{"name":"Center for Vascular Biology, University of Connecticut Health Center 1 , Farmington, CT 06030, USA"},{"name":"University of Connecticut Health Center 2 Department of Cell Biology , , Farmington, CT 06030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[2024,5,31]]},"reference":[{"key":"2024053113560947300_DEV202802C1","doi-asserted-by":"publisher","first-page":"1124","DOI":"10.1016\/j.cell.2009.03.025","article-title":"The notch ligands Dll4 and Jagged1 have opposing effects on angiogenesis","volume":"137","author":"Benedito","year":"2009","journal-title":"Cell"},{"key":"2024053113560947300_DEV202802C2","doi-asserted-by":"publisher","first-page":"849","DOI":"10.1161\/CIRCRESAHA.120.317025","article-title":"Tamoxifen-activated CreERT impairs retinal angiogenesis independently of gene deletion","volume":"127","author":"Brash","year":"2020","journal-title":"Circ. Res."},{"key":"2024053113560947300_DEV202802C3","doi-asserted-by":"publisher","first-page":"1337","DOI":"10.1126\/science.1066373","article-title":"A conserved family of prolyl-4-hydroxylases that modify HIF","volume":"294","author":"Bruick","year":"2001","journal-title":"Science"},{"key":"2024053113560947300_DEV202802C4","doi-asserted-by":"publisher","first-page":"1565","DOI":"10.1038\/nprot.2009.187","article-title":"Quantification of oxygen-induced retinopathy in the mouse: a model of vessel loss, vessel regrowth and pathological angiogenesis","volume":"4","author":"Connor","year":"2009","journal-title":"Nat. Protoc."},{"key":"2024053113560947300_DEV202802C5","doi-asserted-by":"publisher","first-page":"4781","DOI":"10.1158\/0008-5472.CAN-13-0566","article-title":"PanIN-specific regulation of Wnt signaling by HIF2alpha during early pancreatic tumorigenesis","volume":"73","author":"Criscimanna","year":"2013","journal-title":"Cancer Res."},{"key":"2024053113560947300_DEV202802C6","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1046\/j.1471-4159.2001.00012.x","article-title":"Functional gamma-secretase inhibitors reduce beta-amyloid peptide levels in brain","volume":"76","author":"Dovey","year":"2001","journal-title":"J. Neurochem."},{"key":"2024053113560947300_DEV202802C7","doi-asserted-by":"publisher","first-page":"dev175117","DOI":"10.1242\/dev.175117","article-title":"Developmental vascular pruning in neonatal mouse retinas is programmed by the astrocytic oxygen-sensing mechanism","volume":"146","author":"Duan","year":"2019","journal-title":"Development"},{"key":"2024053113560947300_DEV202802C8","doi-asserted-by":"publisher","first-page":"1881","DOI":"10.1016\/j.ajpath.2010.12.016","article-title":"Prolyl hydroxylase domain protein 2 (PHD2) mediates oxygen-induced retinopathy in neonatal mice","volume":"178","author":"Duan","year":"2011","journal-title":"Am. J. Pathol."},{"key":"2024053113560947300_DEV202802C9","doi-asserted-by":"publisher","first-page":"1240","DOI":"10.1016\/j.ajpath.2013.12.014","article-title":"Hematological, hepatic, and retinal phenotypes in mice deficient for prolyl hydroxylase domain proteins in the liver","volume":"184","author":"Duan","year":"2014","journal-title":"Am. J. Pathol."},{"key":"2024053113560947300_DEV202802C10","doi-asserted-by":"publisher","first-page":"e84736","DOI":"10.1371\/journal.pone.0084736","article-title":"Hypoxia inducible factor-2alpha regulates the development of retinal astrocytic network by maintaining adequate supply of astrocyte progenitors","volume":"9","author":"Duan","year":"2014","journal-title":"PLoS ONE"},{"key":"2024053113560947300_DEV202802C11","doi-asserted-by":"publisher","first-page":"17608","DOI":"10.1038\/s41598-017-17962-2","article-title":"Retinal angiogenesis regulates astrocytic differentiation in neonatal mouse retinas by oxygen dependent mechanisms","volume":"7","author":"Duan","year":"2017","journal-title":"Sci. Rep."},{"key":"2024053113560947300_DEV202802C12","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1016\/S0092-8674(01)00507-4","article-title":"C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation","volume":"107","author":"Epstein","year":"2001","journal-title":"Cell"},{"key":"2024053113560947300_DEV202802C13","doi-asserted-by":"publisher","first-page":"27280","DOI":"10.1038\/srep27280","article-title":"HIF-1alpha and HIF-2alpha induced angiogenesis in gastrointestinal vascular malformation and reversed by thalidomide","volume":"6","author":"Feng","year":"2016","journal-title":"Sci. Rep."},{"key":"2024053113560947300_DEV202802C14","doi-asserted-by":"publisher","first-page":"221","DOI":"10.1016\/S1050-1738(03)00082-3","article-title":"Hey genes in cardiovascular development","volume":"13","author":"Fischer","year":"2003","journal-title":"Trends Cardiovasc. Med."},{"key":"2024053113560947300_DEV202802C15","doi-asserted-by":"publisher","first-page":"1513","DOI":"10.1101\/gad.5.9.1513","article-title":"Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice","volume":"5","author":"Friedrich","year":"1991","journal-title":"Genes Dev."},{"key":"2024053113560947300_DEV202802C16","first-page":"522","article-title":"Development of the mouse retinal vasculature: angiogenesis versus vasculogenesis","volume":"43","author":"Fruttiger","year":"2002","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"2024053113560947300_DEV202802C17","doi-asserted-by":"publisher","first-page":"688","DOI":"10.1093\/embo-reports\/kvf124","article-title":"A gamma-secretase inhibitor blocks Notch signaling in vivo and causes a severe neurogenic phenotype in zebrafish","volume":"3","author":"Geling","year":"2002","journal-title":"EMBO Rep."},{"key":"2024053113560947300_DEV202802C18","doi-asserted-by":"publisher","first-page":"1163","DOI":"10.1083\/jcb.200302047","article-title":"VEGF guides angiogenic sprouting utilizing endothelial tip cell filopodia","volume":"161","author":"Gerhardt","year":"2003","journal-title":"J. Cell Biol."},{"key":"2024053113560947300_DEV202802C19","doi-asserted-by":"publisher","first-page":"1253","DOI":"10.1161\/CIRCULATIONAHA.116.023027","article-title":"Loss of endothelial CXCR7 impairs vascular homeostasis and cardiac remodeling after myocardial infarction: implications for cardiovascular drug discovery","volume":"135","author":"Hao","year":"2017","journal-title":"Circulation"},{"key":"2024053113560947300_DEV202802C20","doi-asserted-by":"publisher","first-page":"776","DOI":"10.1038\/nature05571","article-title":"Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis","volume":"445","author":"Hellstr\u00f6m","year":"2007","journal-title":"Nature"},{"key":"2024053113560947300_DEV202802C21","doi-asserted-by":"publisher","first-page":"39792","DOI":"10.1074\/jbc.M206955200","article-title":"Sequence determinants in hypoxia-inducible factor-1\u03b1 for hydroxylation by the prolyl hydroxylases PHD1, PHD2, and PHD3","volume":"277","author":"Huang","year":"2002","journal-title":"J. Biol. Chem."},{"key":"2024053113560947300_DEV202802C22","doi-asserted-by":"publisher","first-page":"13459","DOI":"10.1073\/pnas.192342099","article-title":"Biochemical purification and pharmacological inhibition of a mammalian prolyl hydroxylase acting on hypoxia-inducible factor","volume":"99","author":"Ivan","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2024053113560947300_DEV202802C23","doi-asserted-by":"publisher","first-page":"943","DOI":"10.1038\/ncb2103","article-title":"Endothelial cells dynamically compete for the tip cell position during angiogenic sprouting","volume":"12","author":"Jakobsson","year":"2010","journal-title":"Nat. Cell Biol."},{"key":"2024053113560947300_DEV202802C24","doi-asserted-by":"publisher","first-page":"dev200030","DOI":"10.1242\/dev.200030","article-title":"Oxygen-sensing mechanisms in development and tissue repair","volume":"148","author":"Jiang","year":"2021","journal-title":"Development"},{"key":"2024053113560947300_DEV202802C25","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1007\/BF01680633","article-title":"Griffonia simplicifolia lectins bind specifically to endothelial cells and some epithelial cells in mouse tissues","volume":"19","author":"Laitinen","year":"1987","journal-title":"Histochem. J."},{"key":"2024053113560947300_DEV202802C26","doi-asserted-by":"publisher","first-page":"3219","DOI":"10.1073\/pnas.0611206104","article-title":"Delta-like ligand 4 (Dll4) is induced by VEGF as a negative regulator of angiogenic sprouting","volume":"104","author":"Lobov","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2024053113560947300_DEV202802C27","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1038\/nn.2467","article-title":"A robust and high-throughput Cre reporting and characterization system for the whole mouse brain","volume":"13","author":"Madisen","year":"2010","journal-title":"Nat. Neurosci."},{"key":"2024053113560947300_DEV202802C28","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1038\/20459","article-title":"The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis","volume":"399","author":"Maxwell","year":"1999","journal-title":"Nature"},{"key":"2024053113560947300_DEV202802C29","doi-asserted-by":"publisher","first-page":"dev199418","DOI":"10.1242\/dev.199418","article-title":"Environmental oxygen regulates astrocyte proliferation to guide angiogenesis during retinal development","volume":"148","author":"Perelli","year":"2021","journal-title":"Development"},{"key":"2024053113560947300_DEV202802C30","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1007\/s10456-021-09816-3","article-title":"PFKFB3 gene deletion in endothelial cells inhibits intraplaque angiogenesis and lesion formation in a murine model of venous bypass grafting","volume":"25","author":"Perrotta","year":"2022","journal-title":"Angiogenesis"},{"key":"2024053113560947300_DEV202802C31","doi-asserted-by":"publisher","first-page":"905","DOI":"10.1073\/pnas.92.3.905","article-title":"Vascular endothelial growth factor\/vascular permeability factor expression in a mouse model of retinal neovascularization","volume":"92","author":"Pierce","year":"1995","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2024053113560947300_DEV202802C32","doi-asserted-by":"publisher","first-page":"3807","DOI":"10.1172\/JCI126655","article-title":"Roles of HIFs and VEGF in angiogenesis in the retina and brain","volume":"129","author":"Rattner","year":"2019","journal-title":"J. Clin. Invest."},{"key":"2024053113560947300_DEV202802C33","doi-asserted-by":"publisher","first-page":"2435","DOI":"10.1002\/dvdy.22376","article-title":"The TGFbeta type II receptor plays a critical role in the endothelial cells during cardiac development","volume":"239","author":"Robson","year":"2010","journal-title":"Dev. Dyn."},{"key":"2024053113560947300_DEV202802C34","first-page":"4010","article-title":"Hypoxia-inducible factor-1alpha is a positive factor in solid tumor growth","volume":"60","author":"Ryan","year":"2000","journal-title":"Cancer Res."},{"key":"2024053113560947300_DEV202802C35","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1146\/annurev-pharmtox-010818-021637","article-title":"Pharmacologic targeting of hypoxia-inducible factors","volume":"59","author":"Semenza","year":"2019","journal-title":"Annu. Rev. Pharmacol. Toxicol."},{"key":"2024053113560947300_DEV202802C36","doi-asserted-by":"publisher","first-page":"16002","DOI":"10.1038\/ncomms16002","article-title":"The endothelial transcription factor ERG mediates Angiopoietin-1-dependent control of Notch signalling and vascular stability","volume":"8","author":"Shah","year":"2017","journal-title":"Nat. Commun."},{"key":"2024053113560947300_DEV202802C37","first-page":"723","article-title":"Expression and function of Delta-like ligand 4 in a rat model of retinopathy of prematurity","volume":"8","author":"Shi","year":"2013","journal-title":"Neural Regen. Res."},{"key":"2024053113560947300_DEV202802C38","doi-asserted-by":"publisher","first-page":"469","DOI":"10.1182\/blood-2008-12-193581","article-title":"Endothelial deletion of hypoxia-inducible factor-2alpha (HIF-2alpha) alters vascular function and tumor angiogenesis","volume":"114","author":"Skuli","year":"2009","journal-title":"Blood"},{"key":"2024053113560947300_DEV202802C39","doi-asserted-by":"publisher","first-page":"1427","DOI":"10.1172\/JCI57322","article-title":"Endothelial HIF-2alpha regulates murine pathological angiogenesis and revascularization processes","volume":"122","author":"Skuli","year":"2012","journal-title":"J. Clin. Invest."},{"key":"2024053113560947300_DEV202802C40","doi-asserted-by":"publisher","first-page":"3225","DOI":"10.1073\/pnas.0611177104","article-title":"The Notch ligand Delta-like 4 negatively regulates endothelial tip cell formation and vessel branching","volume":"104","author":"Suchting","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2024053113560947300_DEV202802C41","doi-asserted-by":"publisher","first-page":"178","DOI":"10.1161\/01.HYP.0000251360.40838.0f","article-title":"Prolyl hydroxylase domain 2 protein suppresses hypoxia-induced endothelial cell proliferation","volume":"49","author":"Takeda","year":"2007","journal-title":"Hypertension"},{"key":"2024053113560947300_DEV202802C42","doi-asserted-by":"publisher","first-page":"8336","DOI":"10.1128\/MCB.00425-06","article-title":"Placental but not heart defects are associated with elevated hypoxia-inducible factor alpha levels in mice lacking prolyl hydroxylase domain protein 2","volume":"26","author":"Takeda","year":"2006","journal-title":"Mol. Cell. Biol."},{"key":"2024053113560947300_DEV202802C43","doi-asserted-by":"publisher","first-page":"686","DOI":"10.1016\/j.ajpath.2013.11.032","article-title":"Improved vascular survival and growth in the mouse model of hindlimb ischemia by a remote signaling mechanism","volume":"184","author":"Takeda","year":"2014","journal-title":"Am. J. Pathol."},{"key":"2024053113560947300_DEV202802C44","doi-asserted-by":"publisher","first-page":"485","DOI":"10.1016\/j.ccr.2004.09.026","article-title":"Loss of HIF-1alpha in endothelial cells disrupts a hypoxia-driven VEGF autocrine loop necessary for tumorigenesis","volume":"6","author":"Tang","year":"2004","journal-title":"Cancer Cell"},{"key":"2024053113560947300_DEV202802C45","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1101\/gad.11.1.72","article-title":"Endothelial PAS domain protein 1 (EPAS1), a transcription factor selectively expressed in endothelial cells","volume":"11","author":"Tian","year":"1997","journal-title":"Genes Dev."},{"key":"2024053113560947300_DEV202802C46","doi-asserted-by":"publisher","first-page":"1230","DOI":"10.1074\/jbc.270.3.1230","article-title":"Purification and characterization of hypoxia-inducible factor 1","volume":"270","author":"Wang","year":"1995","journal-title":"J. Biol. Chem."},{"key":"2024053113560947300_DEV202802C47","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1038\/nature09002","article-title":"Ephrin-B2 controls VEGF-induced angiogenesis and lymphangiogenesis","volume":"465","author":"Wang","year":"2010","journal-title":"Nature"},{"key":"2024053113560947300_DEV202802C48","doi-asserted-by":"publisher","DOI":"10.1101\/2023.04.18.537330","article-title":"Monocyte-derived Dll4 is a novel contributor to persistent systemic inflammation in HIV patients","author":"Wang","year":"2023","journal-title":"bioRxiv"},{"key":"2024053113560947300_DEV202802C49","doi-asserted-by":"publisher","first-page":"E3910","DOI":"10.1073\/pnas.1309276110","article-title":"Nrf2 acts cell-autonomously in endothelium to regulate tip cell formation and vascular branching","volume":"110","author":"Wei","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2024053113560947300_DEV202802C50","doi-asserted-by":"publisher","first-page":"1855","DOI":"10.1242\/dev.01732","article-title":"Stabilization of the retinal vascular network by reciprocal feedback between blood vessels and astrocytes","volume":"132","author":"West","year":"2005","journal-title":"Development"},{"key":"2024053113560947300_DEV202802C51","doi-asserted-by":"publisher","first-page":"931","DOI":"10.1182\/blood-2005-03-1000","article-title":"Up-regulation of the Notch ligand Delta-like 4 inhibits VEGF-induced endothelial cell function","volume":"107","author":"Williams","year":"2006","journal-title":"Blood"},{"key":"2024053113560947300_DEV202802C52","doi-asserted-by":"publisher","first-page":"779","DOI":"10.1016\/j.devcel.2020.01.016","article-title":"Sphingosine 1-phosphate receptor signaling establishes AP-1 gradients to allow for retinal endothelial cell specialization","volume":"52","author":"Yanagida","year":"2020","journal-title":"Dev. Cell"},{"key":"2024053113560947300_DEV202802C53","doi-asserted-by":"publisher","first-page":"e27385","DOI":"10.1371\/journal.pone.0027385","article-title":"A computational tool for quantitative analysis of vascular networks","volume":"6","author":"Zudaire","year":"2011","journal-title":"PLoS ONE"}],"container-title":["Development"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.biologists.com\/dev\/article-pdf\/doi\/10.1242\/dev.202802\/3427562\/dev202802.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/journals.biologists.com\/dev\/article-pdf\/doi\/10.1242\/dev.202802\/3467632\/dev202802.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/journals.biologists.com\/dev\/article-pdf\/doi\/10.1242\/dev.202802\/3467632\/dev202802.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,31]],"date-time":"2024-05-31T09:56:29Z","timestamp":1717149389000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.biologists.com\/dev\/article\/151\/11\/dev202802\/352237\/Endothelial-HIF2-suppresses-retinal-angiogenesis"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,31]]},"references-count":53,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2024,6,1]]}},"URL":"https:\/\/doi.org\/10.1242\/dev.202802","relation":{},"ISSN":["0950-1991","1477-9129"],"issn-type":[{"value":"0950-1991","type":"print"},{"value":"1477-9129","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,31]]},"article-number":"dev202802"}}