Synergistic Suppression of Choroidal Neovascularization by Cavtratin and Aflibercept via Inhibition of the eNOS Pathway

Choroidal neovascularization (CNV) is a key pathological feature of exudative age-related macular degeneration (AMD), leading to severe vision loss. Despite anti-vascular endothelial growth factor (anti-VEGF) therapies being the first-line treatment for neovascularization, their long-term application faces challenges including treatment insensitivity and drug resistance. This study aims to investigate the role of Caveolin-1 (Cav-1) in CNV pathogenesis and evaluate the therapeutic potential of Cavtratin, a Cav-1 scaffolding domain-targeting peptide, alone and in combination with Aflibercept. A laser-induced CNV model in aged mice and VEGF-stimulated human umbilical vein endothelial cells (HUVECs) were used to assess Cav-1 expression dynamics and its interaction with endothelial nitric oxide synthase (eNOS). The effects of Cavtratin on angiogenesis were evaluated using tube formation assays, choroidal sprouting assays, and fluorescein angiography. Western blot and immunofluorescence staining were employed to analyze changes in molecular expression, localization, and inflammatory responses. The efficacy of Cavtratin-Aflibercept combination therapy was examined. Cav-1 and eNOS were significantly upregulated during CNV progression (<i>p</i> &lt; 0.001). Cavtratin effectively inhibited tube formation in HUVECs, suppressed choroidal sprouting <i>ex vivo</i>, and reduced CNV leakage <i>in vivo</i> (<i>p</i> &lt; 0.01). Mechanistically, Cavtratin suppressed eNOS phosphorylation and enhanced the anti-angiogenic effects of Aflibercept (<i>p</i> &lt; 0.001). The combination therapy led to greater CNV inhibition, reduced inflammation, and allowed for a lower Aflibercept dosage while maintaining therapeutic efficacy. Cavtratin combined with Aflibercept can effectively enhance anti-angiogenic efficacy and reduce inflammatory responses. Targeting the Cav-1/eNOS axis with Cavtratin provides a novel strategy to complement the limitations of anti-VEGF therapy. The synergistic effects of Cavtratin and Aflibercept suggest a promising approach to overcoming treatment resistance and improving clinical outcomes in CNV management.