title: Myeloperoxidase generates 5-chlorouracil in human atherosclerotic tissue: a potential pathway for somatic mutagenesis by macrophages.
abstract: Somatic mutations induced by oxidative damage of DNA might play important roles in atherogenesis. However, the underlying mechanisms remain poorly understood. Myeloperoxidase, a heme protein expressed by select populations of artery wall macrophages, initiates one potentially mutagenic pathway by generating hypochlorous acid. This potent chlorinating agent reacts rapidly with primary amines to yield long-lived, selectively reactive N-chloramines. In the current studies, we demonstrate that myeloperoxidase produced by human macrophages differentiated in the presence of granulocyte macrophage colony-stimulating factor generates 5-chlorouracil, a mutagenic thymine analog. The primary amine taurine fails to block the reaction, suggesting that N-haloamines produced by macrophages might oxidize uracil. Model system studies demonstrated that N-chloramines convert uracil to 5-chlorouracil. Interestingly, the tertiary amine nicotine dramatically enhances uracil chlorination, suggesting that cigarette smoke might promote nucleobase oxidation by N-chloramines. To look for evidence that myeloperoxidase promotes uracil oxidation in vivo, we measured 5-chlorouracil levels in human aortic tissue, using isotope dilution gas chromatography-mass spectrometry. The level of 5-chlorouracil was 10-fold higher in atherosclerotic aortic tissue obtained during vascular surgery than in normal aortic tissue, suggesting that halogenated nucleobases produced by macrophages might contribute to atherogenesis. Because 5-chlorouracil can be incorporated into nuclear DNA, our observations raise the possibility that halogenation reactions initiated by phagocytes provide one pathway for mutagenesis, phenotypic modulation, and cytotoxicity during atherogenesis.
authors.affiliation: Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors.forename: Junko
authors.initials: J
authors.lastname: Takeshita
authors.forename: Jaeman
authors.initials: J
authors.lastname: Byun
authors.forename: Thomas Q
authors.initials: TQ
authors.lastname: Nhan
authors.forename: David K
authors.initials: DK
authors.lastname: Pritchard
authors.forename: Subramaniam
authors.initials: S
authors.lastname: Pennathur
authors.forename: Steven M
authors.initials: SM
authors.lastname: Schwartz
authors.forename: Alan
authors.initials: A
authors.lastname: Chait
authors.forename: Jay W
authors.initials: JW
authors.lastname: Heinecke
mesh_terms: D000975:Antioxidants / Q000737:chemistry; D001011:Aorta / Q000473:pathology; D050197:Atherosclerosis / Q000378:metabolism*; D002712:Chlorides / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D008401:Gas Chromatography-Mass Spectrometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D006418:Heme / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008264:Macrophages / Q000378:metabolism*; D008956:Models, Chemical; D009000:Monocytes / Q000378:metabolism; D016296:Mutagenesis; D009154:Mutation; D009538:Nicotine / Q000737:chemistry; D009195:Peroxidase / Q000378:metabolism*; D010641:Phenotype; D013696:Temperature; D013941:Thymine / Q000737:chemistry; D013997:Time Factors; D014498:Uracil / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
chemical_list: D000975:Antioxidants; D002712:Chlorides; D006418:Heme; D014498:Uracil; D009538:Nicotine; C010889:5-chlorouracil; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D004247:DNA; D009195:Peroxidase; D013941:Thymine