Cytochrome P450 Oxidase 2J Inhibition Suppresses Choroidal Neovascularization

Abstract

Purpose : Choroidal neovascularization (CNV) in age-related macular degeneration (AMD) can cause blindness. Increased dietary intake of ω-3 long-chain polyunsaturated fatty acids (LCPUFAs) reduces CNV. ω-3 LCPUFA metabolites from most metabolic pathways (COX, LOX) are anti-angiogenic, but ω-3 LCPUFA metabolites of cytochrome P450 oxidase (CYP)2J, promote angiogenesis. We hypothesized that inhibition of CYP2J activity would augment the protective effects of ω-3 LCPUFAs on CNV.

Methods : We investigated the effects of the CYP2J inhibitor (Flunarizine) in the laser-induced CNV mouse model. The plasma levels of CYP2J metabolites were determined by liquid chromatography and tandem mass spectroscopy. Mouse choroidal and aortic explant sprouting assays were used to investigate the effects of CYP2J inhibition and CYP2J metabolites on angiogenesis ex vivo.

Results : CNV was increased in Tie2-driven CYP2J2-overexpressing mice, associated with increased plasma epoxide: diol ratio (n ≥ 25, both genders, p < 0.001). Inhibiting CYP2J activity with flunarizine augmented the protective effects of ω-3 LCPUFAs on CNV by 20% (n ≥ 26, both genders, p < 0.05). In Tie2-driven CYP2J2-overexpressing mice fed a ω-3 LCPUFA diet, flunarizine suppressed choroidal neovascularization by 39% (n ≥ 33, both genders, p < 0.001) and reduced the plasma levels of CYP2J2 metabolites. In addition, the combination therapy of Flunaridine and Montelukast (CYP2C inhibitor) enhanced the prevention of CNV. The proangiogenic role of CYP2J2 was corroborated in choroid and aortic sprouting assays. Exposure to the ω-3 LCPUFA CYP2J metabolite, 19,20-epoxydocosapentaenoic acid, reversed the suppression of angiogenesis ex vivo by the CYP2J inhibitor flunarizine.

Conclusions : CYP2J Inhibition augmented the protective effects of ω-3 LCPUFAs on pathological choroidal angiogenesis. Flunarizine suppressed angiogenesis by inhibiting CYP2J activity, and Montelukast enhanced the effect. CYP2 inhibition may be a viable approach to inhibit CNV in AMD.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.