Abstract 380: Notch4 Uncouples Endothelial Nitric Oxide Synthase Leading to Arteriovenous Malformations

Abstract

Notch is distinctively expressed in arterial but not venous endothelial cells. Notch signaling regulates arteriovenous (AV) specification, and mutations in Notch signaling lead to AV malformation (AVM) in mice. AVMs are characterized by abnormal AV shunts that displace capillaries. Mechanisms underlying AVM pathogenesis remain poorly understood, hindering therapeutic development. We reported that endothelial expression of constitutively active Notch4 (Notch4*) in mice initiates AVMs de novo through enlargement of microvessels without an increase in endothelial cell number or proliferation. Here, we hypothesized that Notch4* disrupts endothelial nitric oxide synthase (eNOS) signaling and vascular tone, thereby permitting vessel enlargement and AV shunting. We show that arteries isolated from Notch4* mutant mice exhibited decreased arterial tone compared to controls, suggesting that Notch4* impaired vascular tone. Administering the NOS inhibitor N G -nitro-L-arginine abolished Notch4*-mediated vascular tone impairment. Deletion of the eNOS gene and administration of the NOS inhibitor N G -nitro-L-arginine in the Notch4* mutant mice attenuated Notch4*-induced AVM initiation, measured by decreased AV shunt diameter, delayed AV shunting, reduced hemorrhage, pathological lesions, and improved survival, suggesting that eNOS is essential for Notch4* action. In addition, uncoupled eNOS-derived superoxide production was elevated in the Notch4* mice. Our results show that inhibition of eNOS signaling attenuates Notch4*-mediated AVM formation. Furthermore, Notch4* impairs eNOS activity, leading to superoxide production, which results in arterial dysfunction and AV shunt formation.