Yan Gong1, Yohei Tomita2, Matthew L Edin3, Anli Ren4, Minji Ko2, Jay Yang2, Edward Bull2, Darryl C Zeldin3, Ann Hellström5, Zhongjie Fu2, Lois E H Smith6. 1. Department of Biological Repositories, Zhongnan Hospital of Wuhan University, Wuhan, China; Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA. 2. Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA. 3. Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC, USA. 4. Department of Biological Repositories, Zhongnan Hospital of Wuhan University, Wuhan, China; Department of Ophthalmology, Zhongnan Hospital of Wuhan University, Wuhan, China. 5. Institute of Neuroscience and Physiology, Sahlgrenska Academy, Gothenburg University, Göteborg, Sweden. 6. Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA. Electronic address: lois.smith@childrens.harvard.edu.
Abstract
INTRODUCTION: Choroidal neovascularization (CNV) in age-related macular degeneration (AMD) leads to blindness. It has been widely reported that increased intake of ω-3 long-chain polyunsaturated fatty acids (LCPUFA) diets reduce CNV. Of the three major pathways metabolizing ω-3 (and ω-6 LCPUFA), the cyclooxygenase and lipoxygenase pathways generally produce pro-angiogenic metabolites from ω-6 LCPUFA and anti-angiogenic ones from ω-3 LCPUFA. Howevehr, cytochrome P450 oxidase (CPY) 2C produces pro-angiogenic metabolites from both ω-6 and ω-3 LCPUFA. The effects of CYP2J2 products on ocular neovascularization are still unknown. Understanding how each metabolic pathway affects the protective effect of ω-3 LCPUFA on retinal neovascularization may lead to therapeutic interventions. OBJECTIVES: To investigate the effects of LCPUFA metabolites through CYP2J2 pathway and CYP2J2 regulation on CNV both in vivo and ex vivo. METHODS: The impact of CYP2J2 overexpression and inhibition on neovascularization in the laser-induced CNV mouse model was assessed. The plasma levels of CYP2J2 metabolites were measured by liquid chromatography and tandem mass spectroscopy. The choroidal explant sprouting assay was used to investigate the effects of CYP2J2 inhibition and specific LCPUFA CYP2J2 metabolites on angiogenesis ex vivo. RESULTS: CNV was exacerbated in Tie2-Cre CYP2J2-overexpressing mice and was associated with increased levels of plasma docosahexaenoic acids. Inhibiting CYP2J2 activity with flunarizine decreased CNV in both ω-6 and ω-3 LCPUFA-fed wild-type mice. In Tie2-Cre CYP2J2-overexpressing mice, flunarizine suppressed CNV by 33 % and 36 % in ω-6, ω-3 LCPUFA diets, respectively, and reduced plasma levels of CYP2J2 metabolites. The pro-angiogenic role of CYP2J2 was corroborated in the choroidal explant sprouting assay. Flunarizine attenuated ex vivo choroidal sprouting, and 19,20-EDP, a ω-3 LCPUFA CYP2J2 metabolite, increased sprouting. The combined inhibition of CYP2J2 with flunarizine and CYP2C8 with montelukast further enhanced CNV suppression via tumor necrosis factor-α suppression. CONCLUSIONS: CYP2J2 inhibition augmented the inhibitory effect of ω-3 LCPUFA on CNV. Flunarizine suppressed pathological choroidal angiogenesis, and co-treatment with montelukast inhibiting CYP2C8 further enhanced the effect. CYP2 inhibition might be a viable approach to suppress CNV in AMD.
INTRODUCTION: Choroidal neovascularization (CNV) in age-related macular degeneration (AMD) leads to blindness. It has been widely reported that increased intake of ω-3 long-chain polyunsaturated fatty acids (LCPUFA) diets reduce CNV. Of the three major pathways metabolizing ω-3 (and ω-6 LCPUFA), the cyclooxygenase and lipoxygenase pathways generally produce pro-angiogenic metabolites from ω-6 LCPUFA and anti-angiogenic ones from ω-3 LCPUFA. Howevehr, cytochrome P450 oxidase (CPY) 2C produces pro-angiogenic metabolites from both ω-6 and ω-3 LCPUFA. The effects of CYP2J2 products on ocular neovascularization are still unknown. Understanding how each metabolic pathway affects the protective effect of ω-3 LCPUFA on retinal neovascularization may lead to therapeutic interventions. OBJECTIVES: To investigate the effects of LCPUFA metabolites through CYP2J2 pathway and CYP2J2 regulation on CNV both in vivo and ex vivo. METHODS: The impact of CYP2J2 overexpression and inhibition on neovascularization in the laser-induced CNV mouse model was assessed. The plasma levels of CYP2J2 metabolites were measured by liquid chromatography and tandem mass spectroscopy. The choroidal explant sprouting assay was used to investigate the effects of CYP2J2 inhibition and specific LCPUFA CYP2J2 metabolites on angiogenesis ex vivo. RESULTS: CNV was exacerbated in Tie2-Cre CYP2J2-overexpressing mice and was associated with increased levels of plasma docosahexaenoic acids. Inhibiting CYP2J2 activity with flunarizine decreased CNV in both ω-6 and ω-3 LCPUFA-fed wild-type mice. In Tie2-Cre CYP2J2-overexpressing mice, flunarizine suppressed CNV by 33 % and 36 % in ω-6, ω-3 LCPUFA diets, respectively, and reduced plasma levels of CYP2J2 metabolites. The pro-angiogenic role of CYP2J2 was corroborated in the choroidal explant sprouting assay. Flunarizine attenuated ex vivo choroidal sprouting, and 19,20-EDP, a ω-3 LCPUFA CYP2J2 metabolite, increased sprouting. The combined inhibition of CYP2J2 with flunarizine and CYP2C8 with montelukast further enhanced CNV suppression via tumor necrosis factor-α suppression. CONCLUSIONS: CYP2J2 inhibition augmented the inhibitory effect of ω-3 LCPUFA on CNV. Flunarizine suppressed pathological choroidal angiogenesis, and co-treatment with montelukast inhibiting CYP2C8 further enhanced the effect. CYP2 inhibition might be a viable approach to suppress CNV in AMD.
Authors: Matthew L Edin; Zhongjing Wang; J Alyce Bradbury; Joan P Graves; Fred B Lih; Laura M DeGraff; Julie F Foley; Robert Torphy; Oline K Ronnekleiv; Kenneth B Tomer; Craig R Lee; Darryl C Zeldin Journal: FASEB J Date: 2011-06-22 Impact factor: 5.191
Authors: Alexander J Nelson; Daniel J Stephenson; Christopher L Cardona; Xiaoyong Lei; Abdulaziz Almutairi; Tayleur D White; Ying G Tusing; Margaret A Park; Suzanne E Barbour; Charles E Chalfant; Sasanka Ramanadham Journal: J Lipid Res Date: 2019-12-09 Impact factor: 5.922
Authors: Yan Gong; Jie Li; Ye Sun; Zhongjie Fu; Chi-Hsiu Liu; Lucy Evans; Katherine Tian; Nicholas Saba; Thomas Fredrick; Peyton Morss; Jing Chen; Lois E H Smith Journal: PLoS One Date: 2015-07-10 Impact factor: 3.240
Authors: Johanna M Colijn; Gabriëlle H S Buitendijk; Elena Prokofyeva; Dalila Alves; Maria L Cachulo; Anthony P Khawaja; Audrey Cougnard-Gregoire; Bénédicte M J Merle; Christina Korb; Maja G Erke; Alain Bron; Eleftherios Anastasopoulos; Magda A Meester-Smoor; Tatiana Segato; Stefano Piermarocchi; Paulus T V M de Jong; Johannes R Vingerling; Fotis Topouzis; Catherine Creuzot-Garcher; Geir Bertelsen; Norbert Pfeiffer; Astrid E Fletcher; Paul J Foster; Rufino Silva; Jean-François Korobelnik; Cécile Delcourt; Caroline C W Klaver Journal: Ophthalmology Date: 2017-07-14 Impact factor: 12.079
Authors: Zhuo Shao; Mollie Friedlander; Christian G Hurst; Zhenghao Cui; Dorothy T Pei; Lucy P Evans; Aimee M Juan; Houda Tahiri; Houda Tahir; François Duhamel; Jing Chen; Przemyslaw Sapieha; Sylvain Chemtob; Jean-Sébastien Joyal; Lois E H Smith Journal: PLoS One Date: 2013-07-26 Impact factor: 3.240