Endothelium-Intrinsic Requirement for Hif-2 α During Vascular Development

BACKGROUND: The development of the vascular system is a complex process that involves communications among multiple cell types. As such, it is important to understand whether a specific gene regulates vascular development directly from within the vascular system or indirectly from nonvascular cells. Hypoxia-inducible factor-2alpha (Hif-2alpha, or endothelial PAS protein-1 [EPAS-1]) is required for vascular development in mice, but it is not clear whether its requirement resides directly in endothelial cells. METHODS AND RESULTS: To address this issue, we expressed Hif-2alpha cDNA in the vascular endothelium of Hif-2alpha-/- embryos by an embryonic stem (ES) cell-mediated transgenic approach and assessed whether endothelium-specific reexpression of Hif-2alpha could rescue vascular development. Here we report that although ES cell-derived Hif-2alpha-/- embryos developed severe vascular defects by embryonic day (E) 11.5 and died in utero before E12.5, endothelium-specific expression of Hif-2alpha cDNA restored normal vascular development at all stages examined (up to E14.5) and allowed Hif-2alpha-/- embryos to survive at a frequency comparable to that of Hif-2alpha+/- embryos. Furthermore, we found that Tie-2 expression was significantly reduced in Hif-2alpha-/- mutants but was restored by Hif-2alpha cDNA expression. CONCLUSIONS: These data demonstrate an intrinsic requirement for Hif-2alpha by endothelial cells and imply that hypoxia may control endothelial functions directly via Hif-2alpha-regulated Tie-2 expression.