title: Cytochrome P450 Oxidase 2C Inhibition Adds to ω-3 Long-Chain Polyunsaturated Fatty Acids Protection Against Retinal and Choroidal Neovascularization.
abstract: Pathological ocular neovascularization is a major cause of blindness. Increased dietary intake of ω-3 long-chain polyunsaturated fatty acids (LCPUFA) reduces retinal neovascularization and choroidal neovascularization (CNV), but ω-3 LCPUFA metabolites of a major metabolizing pathway, cytochrome P450 oxidase (CYP) 2C, promote ocular pathological angiogenesis. We hypothesized that inhibition of CYP2C activity will add to the protective effects of ω-3 LCPUFA on neovascular eye diseases.
  
  
  
  The mouse models of oxygen-induced retinopathy and laser-induced CNV were used to investigate pathological angiogenesis in the retina and choroid, respectively. The plasma levels of ω-3 LCPUFA metabolites of CYP2C were determined by mass spectroscopy. Aortic ring and choroidal explant sprouting assays were used to investigate the effects of CYP2C inhibition and ω-3 LCPUFA-derived CYP2C metabolic products on angiogenesis ex vivo. We found that inhibition of CYP2C activity by montelukast added to the protective effects of ω-3 LCPUFA on retinal neovascularization and CNV by 30% and 20%, respectively. In CYP2C8-overexpressing mice fed a ω-3 LCPUFA diet, montelukast suppressed retinal neovascularization and CNV by 36% and 39% and reduced the plasma levels of CYP2C8 products. Soluble epoxide hydrolase inhibition, which blocks breakdown and inactivation of CYP2C ω-3 LCPUFA-derived active metabolites, increased oxygen-induced retinopathy and CNV in vivo. Exposure to selected ω-3 LCPUFA metabolites of CYP2C significantly reversed the suppression of both angiogenesis ex vivo and endothelial cell functions in vitro by the CYP2C inhibitor montelukast.
  
  
  
  Inhibition of CYP2C activity adds to the protective effects of ω-3 LCPUFA on pathological retinal neovascularization and CNV.
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Yan
authors.initials: Y
authors.lastname: Gong
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Zhongjie
authors.initials: Z
authors.lastname: Fu
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Matthew L
authors.initials: ML
authors.lastname: Edin
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Chi-Hsiu
authors.initials: CH
authors.lastname: Liu
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Zhongxiao
authors.initials: Z
authors.lastname: Wang
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Shao
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Fredrick
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Saba
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Peyton C
authors.initials: PC
authors.lastname: Morss
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Samuel B
authors.initials: SB
authors.lastname: Burnim
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Steven S
authors.initials: SS
authors.lastname: Meng
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Fred B
authors.initials: FB
authors.lastname: Lih
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Kin Sing Stephen
authors.initials: KS
authors.lastname: Lee
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Elizabeth P
authors.initials: EP
authors.lastname: Moran
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: John Paul
authors.initials: JP
authors.lastname: SanGiovanni
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Ann
authors.initials: A
authors.lastname: Hellström
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Hammock
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.).
authors.forename: Darryl C
authors.initials: DC
authors.lastname: Zeldin
authors.affiliation: From the Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (Y.G., Z.F., C.-H.L., Z.W., Z.S., T.W.F., N.J.S., P.C.M., S.B.B., S.S.M., E.P.M., L.E.H.S.); Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC (M.L.E., F.B.L., D.C.Z.); Department of Entomology and Comprehensive Cancer Center, University of California, Davis (K.S.S.L., B.D.H.); Section on Nutritional Neurosciences in the Laboratory of Membrane Biophysics and Biochemistry, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD (J.P.S.G.); and Department of Ophthalmology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden (A.H.). lois.smith@childrens.harvard.edu.
authors.forename: Lois E H
authors.initials: LE
authors.lastname: Smith
mesh_terms: D000085:Acetates / Q000494:pharmacology*; D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000201:enzymology; D002478:Cells, Cultured; D020256:Choroidal Neovascularization / Q000201:enzymology / Q000235:genetics / Q000503:physiopathology / Q000517:prevention & control*; D003521:Cyclopropanes; D065727:Cytochrome P-450 CYP2C8 / Q000235:genetics / Q000378:metabolism*; D065687:Cytochrome P-450 CYP2C8 Inhibitors / Q000494:pharmacology*; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000187:drug effects / Q000201:enzymology; D015525:Fatty Acids, Omega-3 / Q000378:metabolism / Q000494:pharmacology*; D005838:Genotype; D006801:Humans; D018496:Hyperoxia / Q000150:complications; D007834:Lasers; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018919:Neovascularization, Physiologic / Q000187:drug effects; D010641:Phenotype; D011804:Quinolines / Q000494:pharmacology*; D015861:Retinal Neovascularization / Q000201:enzymology / Q000235:genetics / Q000503:physiopathology / Q000517:prevention & control*; D012178:Retinopathy of Prematurity / Q000201:enzymology / Q000235:genetics / Q000503:physiopathology / Q000517:prevention & control*; D013440:Sulfides; D046509:Tissue Culture Techniques
chemical_list: D000085:Acetates; D020533:Angiogenesis Inhibitors; D003521:Cyclopropanes; D065687:Cytochrome P-450 CYP2C8 Inhibitors; D015525:Fatty Acids, Omega-3; D011804:Quinolines; D013440:Sulfides; C450259:CYP2C8 protein, human; D065727:Cytochrome P-450 CYP2C8; C093875:montelukast
keywords: CYP2C inhibitor; arachidonic acid; choroidal neovascularization; diabetic retinopathy; docosahexaenoic acid