The vascular endothelial growth factor trap aflibercept induces vascular dysfunction and hypertension via attenuation of eNOS/NO signaling in mice.

Abstract

Aflibercept, as a soluble decoy vascular endothelial growth factor receptor,Which has been used as afirst-line monotherapy for cancers.Aflibercept often causes cardiovascular toxicities including hypertension, but the mechanisms underlyingaflibercept-induced hypertension remain unknown. In this study we investigated the effect of short-term and long-term administrationof aflibercept on blood pressure (BP), vascular function, NO bioavailability, oxidative stress and endothelin 1 (ET-1) in mice and cultured endothelial cells. We showed that injection of a single-dose of aflibercept (18.2, 36.4 mg/kg, iv) rapidly and dose-dependently elevated BP in mice. Aflibercept treatment markedly impaired endothelial-dependent relaxation (EDR) andresulted in NADPH oxidases 1 (NOX1)- and NADPH oxidases4 (NOX4)-mediated generation of ROS, decreased theactivation of protein kinase B (Akt) and endothelial nitric oxide synthase (eNOS) concurrently with a reduction in nitric oxide (NO) production and elevation of ET-1levelsin mouse aortas;theseeffects were greatly attenuated by supplementation ofL-arginine (L-arg, 0.5 or 1.0 g/kg, bid, ig) before aflibercept injection. Similar results were observed in L-arg-pretreated cultured endothelial cells, showing markedly decreased ROS accumulation and AKT/eNOS/NO signaling impairment induced by aflibercept. In order to assess the effects of long-term aflibercept on hypertension and to evaluate the beneficial effects ofL-arg supplementation, we administered these two drugs to WT mice for up to 14 days(at an interval of two days). Long-term administration of aflibercept resulted in a sustained increase in BP and a severely impaired EDR, which are associated with NOX1/NOX4-mediated production of ROS, increase in ET-1, inhibition of AKT/eNOS/NO signaling and a decreased expression of cationic amino acid transporter (CAT-1). The effects causedby long-term administration were greatly attenuated by L-arg supplementation in a dose-dependent manner. We conclude that aflibercept leads to vascular dysfunction and hypertension by inhibiting CAT-1/AKT/eNOS/NO signaling, increasing ET-1, and activating NOX1/NOX4-mediated oxidative stress, which can be suppressed by supplementation ofL-arg. Therefore, L-arg could be a potential therapeutic agent for aflibercept-induced hypertension.