DNAアダクトーム解析による未知DNA損傷の構造決定とその生物影響評価

2009 Fiscal Year Annual Research Report

• Project/Area Number: 18101003
• Research Institution: Kyoto University
• Principal Investigator: 松田 知成 Kyoto University, 工学研究科, 准教授 (50273488)
• Co-Investigator(Kenkyū-buntansha): 佐伯 憲一 横浜薬科大学, 薬学部, 准教授 (60254306) ; 高村 岳樹 神奈川工科大学, 工学部, 准教授 (50342910) ; 川西 優喜 大阪府立大学, 産学連携機構, 助教 (70332963)
• Keywords: LC / MS / MS / アダクトーム / DNA損傷 / 突然変異 / 過酸化脂質 / 3-ニトロベンゾアントロン

Research Abstract

まず、DNA付加体の定量研究の成果では、飲酒により生成するDNA付加体の解析を、アルコール依存症患者の血液やaldh2ノックアウトマウスの臓器を用いて進め、アルコール及びアセトアルデヒド代謝にかかわる遺伝子多型によりDNA損傷が有意に影響を受けることを示した。さらに、多くの食品中に含まれるアクリルアミドをラットに投与してDNA付加体の測定を行い、若いラットと大人のラットでその影響にはっきりとした差があることを突き止めた。
大気汚染物質の3-ニトロベンズアントロン(3-NBA)のDNA損傷性に関する研究も進めている。LC/MS/MS法と従来の32P-ポストラベル法を用いてヒト細胞内の3-NBA由来の各種付加体のDNA修復のキネティクスを明らかにした。また3-NBAの付加体とその生体影響の関連を調べる目的でCHL/IU細胞を用いた染色体異常試験を行った。その結果3-NBAは6ng/mlという極めて低濃度から染色体異常を誘発することが明らかとなった。一方で,代謝活性体であるヒドロキシアミノ体やN-アセチルヒドロキシアミノ体では5μg/mLで染色体異常が観測された。この理由は現在調査中である。
また、昨年までの研究で、脂質過酸化反応により生成するDNA付加体が、ヒト組織中に普遍的に存在し、時として非常に高い値を示すことがわかってきた。そこで、シャトルベクターの突然変異検出系を用いて、過酸化脂質により生成するDNA損傷の変異誘発性について検討し、突然変異の誘発を確認した。一方、付加体の修復系を明らかにするため、DNA損傷に結合するタンパク質をSILACという技術を用いて網羅的に解析する方法を確認した。

Research Products

• [Journal Article] Genotoxicity of acrylamide in vitro : Acrylamide is not metabolically activated in standard in vitro systems. (2010)
  • Author(s): Koyama N, Yasui M, Matsuda T, Honma M, et al.
  • Journal Title: Environ Mol Mutagen (in press)
  • Peer Reviewed
• [Journal Article] DNA modifications by the omega-3 lipid peroxidation-derived mutagen 4-oxo-2-hexenal in vitro and their (2010)
  • Author(s): Kawai K, Chou PH, Matsuda T, Kasai H, et al.
  • Journal Title: Chem Res Toxicol 23 Pages: 630-636
  • Peer Reviewed
• [Journal Article] Lipid peroxidation generates a body odor component trans-2-nonenal covalently bound to protein in vivo. (2010)
  • Author(s): Ishino K, Wakita C, Matsuda T, Uchida K, et al.
  • Journal Title: J Biol Chem (in press)
  • Peer Reviewed
• [Journal Article] N2-ethylidene-2'-deoxyguanosine DNA adducts in organs of Aldh2 Knockout mice treated with acetaldehyde inhalation. (2010)
  • Author(s): Oyama, T., Nagayoshi, H., Matsuda, T., Oka, M., Isse, T., Yu, H.S., Phuong, P.T.T., Tanaka, M., Kagawa, N., Kaneko, K., Kawamoto, T
  • Journal Title: Frontiers in Bioscience (in press)
  • Peer Reviewed
• [Journal Article] Genotoxicity of nanoparticles in in vitro micronuclei, in vivo comet and mutation assay systems. (2010)
  • Author(s): Yukari Totsuka, Takashi Higuchi, Toshio Imai, Akiyoshi Nishikawa, Takehiko Nohmi, Tatsuya Kato, Shuich Masuda, Naohide Kinae, Kyoko Hiyoshi, Sayaka Ogo, Masanobu Kawanishi, Takashi Yagi, Takamichi Ichinose, Takashi Sugimura, Keiji Wakabayashi.
  • Journal Title: Particle and fibre toxicology (in pree)
  • Peer Reviewed
• [Journal Article] Establishment of yeast reporter assay systems to detect ligands of thyroid hormone receptors alpha and beta. (2010)
  • Author(s): Kazuhiro Shiizaki, Shota Asai, Shingo Ebata, Masanobu Kawanishi, Takashi Yagi.
  • Journal Title: Toxicology in Vitro (in pree)
  • Peer Reviewed
• [Journal Article] Radiation Dose-Rate Effect on Mutation Induction in Spleen and Liver of gpt delta Mice (2010)
  • Author(s): Naohito Okudaira, Yoshihiko Uehara, Kazuo Fujikawa, Nao Kagawa, Akira Ootsuyama, Toshiyuki Norimura, Ken-ichi Saeki, Takehito Nohmi, Ken-ichi masumura, Tsuneya Matsumoto, Yoichi Oghiso, Kimio Tanaka, Kazuaki Ichinohe, Shingo Nakamura, Satoshi Tanaka, Tesuya Ono
  • Journal Title: Radiation Res 173 Pages: 138-147
  • Peer Reviewed
• [Journal Article] 液体クロマトグラフィータンデム質量分析法を用いたDNA損傷研究法. (2009)
  • Author(s): 松田, 永吉, 椙村, 周
  • Journal Title: J.Mass Spectrom.Soc.Jpn. 57 Pages: 301-304
• [Journal Article] アダクトミクス-DNAおよびタンパク質付加体の網羅的解析. (2009)
  • Author(s): 松田, 足立, 周
  • Journal Title: 実験医学増刊 27 Pages: 2481-2488
• [Journal Article] Quantification of 3-nitrobenzanthrone-DNA adducts using online column-switching HPLC-electrospray tandem mass spectrometry. (2009)
  • Author(s): Gamboa da Costa G, Singh R, Arlt VM, Mirza A, Richards M, Takamura-Enya T, Schmeiser HH, Farmer PB, Phillips DH
  • Journal Title: Chem Res Toxicol 22(11) Pages: 1860-1868
  • Peer Reviewed
• [Journal Article] Isolation and identification of a novel aromatic amine mutagen produced by the Maillard reaction (2009)
  • Author(s): Nishigaki R, Watanabe T, Kajimoto T, Tada A, Takamura-Enya T, Enomoto S, Nukaya H, Terao Y, Muroyama A, Ozeki M, Node M, Hasei T, Totsuka Y, Wakabayashi K
  • Journal Title: Chem Res Toxicol 22(9) Pages: 1588-1593
  • Peer Reviewed
• [Journal Article] H. Translesional DNA synthesis through a C8-guanyl adduct of 2-amino-1-methyl-6-phenylimidazo[4, 5-b]pyridine(PhIP)in Vitro : REV1 inserts dC oppostie the lesion, and DNA polymerase kappa potentially catalyzes extension reaction from the 3'-dC terminus. (2009)
  • Author(s): Fukuda H, Takamura-Enya T, Masuda Y, Nohmi T, Seki C, Kamiya K, Sugimura T, Masutani C, Hanaoka F, Nakagama H.
  • Journal Title: J Biol Chem 248(38) Pages: 25585-25592
  • Peer Reviewed
• [Journal Article] The genotoxic air pollutant 3-nitrobenzanthrone and its reactive metabolite N-hydroxy-3-aminobenzanthrone lack initiating and complete carcinogenic activity in NMRI mouse skin. (2009)
  • Author(s): Scheiser HH, Furstenberger G, Takamura-Enya T, Phillips DH, Arlt VM.
  • Journal Title: Cancer Lett 284(1) Pages: 21-29
  • Peer Reviewed
• [Journal Article] Genotoxicity of 3, 6-dintrobenzo[e]pyrene, a novel mutagen in ambient air and surface soil, in mammalian cells in vitro and in vivo. (2009)
  • Author(s): Masanobu Kawanishi, Tetsushi Watanabe, Soichiro Hagio, Sayaka Ogo, Chiaki Shimohara, Rika Jouchi, Saori Takayama, Tomohiro Hasei, Teruhisa Hirayama, Yoshimitsu Oda, Takashi Yagi
  • Journal Title: Mutagenesis 24(3) Pages: 279-284
  • Peer Reviewed
• [Journal Article] Differential responese of various pharmaceuticals to human estrogen receptors alpha and beta in newly-constructed yeast reporter assays. (2009)
  • Author(s): Wai-Ling Chu, Kazuhiro Shiizaki, Masanobu Kawanishi, Takashi Yagi.
  • Journal Title: Genes and Environment 31(3) Pages: 80-86
  • Peer Reviewed
• [Journal Article] ヒト細胞におけるアミノビフェニルDNA付加体の損傷乗り越えDNA合成を介した突然変異誘発 (2009)
  • Author(s): 澤井知子, 川西優喜, 八木孝司, 高村岳樹
  • Journal Title: 放射線生物研究 44(3) Pages: 274-284
• [Presentation] ALDH2 genotype is critical for DNA adducts formation in mice treated with alcohol and acetaldehyde. (2009)
  • Author(s): Matsuda T, Nagayoshi H, Oka M, et al.
  • Organizer: 10th International Conference on Environmental Mutagens (ICEM)
  • Place of Presentation: Firenze, Italy
  • Year and Date: 20090820-20090825
• [Presentation] Significance of DNA adductome analysis in in vitro micronucleus test (2009)
  • Author(s): Muto S, Kato K, Yamamura E, et al.
  • Organizer: 10th International Conference on Environmental Mutagens (ICEM)
  • Place of Presentation: Firenze, Italy.
  • Year and Date: 20090820-20090825
• [Presentation] Approach to toxicity evaluation of C60 fullerene using several in vitro methods (2009)
  • Author(s): Matsuda S, Matsui S, Matsuda T.
  • Organizer: 10th International Conference on Environmental Mutagens (ICEM)
  • Place of Presentation: Firenze, Italy.
  • Year and Date: 20090820-20090825
• [Presentation] Child-adult differences in evaluation of in vivo genotoxicity of acrylamide. (2009)
  • Author(s): Koyama N, Kimura A, Yasui M, et al
  • Organizer: 10th International Conference on Environmental Mutagens (ICEM)
  • Place of Presentation: Firenze, Italy.
  • Year and Date: 20090820-20090825
• [Presentation] Formation, DNA repair, and TLS of 3-nitrobenzanthrone-derived DNA adducts. (2009)
  • Author(s): Kawanishi M, Nishida H, Ishii H, et al.
  • Organizer: 10th International Conference on Environmental Mutagens (ICEM)
  • Place of Presentation: Firenze, Italy.
  • Year and Date: 20090820-20090825
• [Presentation] Frequent incorporation of formaldehyde derived N2-methyl-2'-deoxyguanosine triphosphate into DNA during DNA synthesis catalyzed by bacterial and mammarian DNA polymerase. (2009)
  • Author(s): Ihara M, Yasui M, Matsui S, Shibutani S, Matsuda T.
  • Organizer: 10th International Conference on Environmental Mutagens (ICEM)
  • Place of Presentation: Firenze, Italy.
  • Year and Date: 20090820-20090825
• [Presentation] Detection of liqid peroxidation-induced DNA adducts caused by 4-oxo-2-nonenal and 4-oxo-2-hexenal in human autopsy tissues. (2009)
  • Author(s): Chou P-H, Kageyama S, Matsuda S, et al.
  • Organizer: 10th International Conference on Environmental Mutagens (ICEM)
  • Place of Presentation: Firenze, Italy.
  • Year and Date: 20090820-20090825
• [Presentation] Oxidative modificomics analysis of cysteine thiols. (2009)
  • Author(s): Adachi J, Kihara K, Matsuda T.
  • Organizer: 10th International Conference on Environmental Mutagens (ICEM)
  • Place of Presentation: Firenze, Italy.
  • Year and Date: 20090820-20090825