Crosstalk Between EnNaC and VEGFR2 as a Novel Mechanism Driving ROS-dependent Angiogenesis

Angiogenesis is essential for vascular repair and is a key therapeutic target in peripheral arterial disease (PAD). VEGF activates VEGF receptor-2 (VEGFR2) within caveolae/lipid rafts (C/LR) microdomains, triggering downstream signaling that drives endothelial cells (EC) angiogenic response. We previously showed that reactive oxygen species (ROS), particularly H 2 O 2 , enhance VEGFR2 phosphorylation and angiogenesis; however, how VEGF rapidly induces ROS to amplify VEGFR2 signaling remains unclear. The epithelial sodium channel (ENaC), whose surface expression is regulated by serum/glucocorticoid-regulated kinase 1 (SGK1), is expressed in ECs (EnNaC) and modulates membrane mechanics and barrier function, but its role in angiogenic signaling is unknown. Here, we show that EC-specific EnNaC knockout mice exhibited markedly impaired blood flow recovery following hindlimb ischemia (33.3% at day 14). In in-vitro, EnNaC knockdown in human ECs significantly suppressed VEGF-induced migration and proliferation. Mechanistically, loss of EnNaC inhibitedrapid VEGF-induced ROS generation (3.4-fold), VEGFR2 tyrosine phosphorylation, and downstream Akt activation. VEGF stimulation induced rapid physical association of EnNaC with VEGFR2, and C/LR fractionation revealed co-localization of both proteins within C/LR microdomains, suggesting spatial signaling crosstalk. Pharmacological inhibition of sodium channels with amiloride or benzamil blocked VEGF-induced EC migration and capillary tube formation without suppressing VEGFR2 phosphorylation or downstream signaling, suggesting that ENaC-dependent angiogenesis is at least partially uncoupled from Na + uptake. In addition, VEGF stimulated SGK1 phosphorylation in ECs, supporting a mechanism whereby VEGF enhances EnNaC membrane stability to facilitate receptor coupling. Collectively, these findings demonstrate that VEGF-induced EnNaC binding to VEGFR2 within C/LR promotes localized ROS production that amplifies VEGFR2 signaling, thereby promoting Na + -dependent angiogenesis after ischemic injury. Thus, endothelial EnNaC represents a previously unrecognized hub linking membrane compartmentalization, VEGF-induced ROS signaling and angiogenesis, providing new mechanistic insight and potential therapeutic target for vascular repair in PAD. This abstract was presented at the American Physiology Summit 2026 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.