ゲノム－核内受容体の相互作用アレイによる化学物質影響評価系の開発

2015 Fiscal Year Annual Research Report

• Project/Area Number: 25660228
• Research Institution: Ehime University
• Principal Investigator: 岩田 久人 愛媛大学, 沿岸環境科学研究センター, 教授 (10271652)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 核内受容体 / 化学物質 / スクリーニング / 種差

Outline of Annual Research Achievements

昨年度までの研究で、野生生物のERを介した反応をスクリーニングする方法が構築できた。一方、バイカルアザラシ（Pusa sibirica）はポリ塩化ビフェニル（PCBs）などの残留性有機汚染物質（POPs）を高蓄積している。これらPCBsは本種の肝臓で、シトクロムP450（CYP）によって水酸化代謝物であるOH-PCBsへと代謝される。そこで今年度は、バイカルアザラシERアイソフォーム（bsERα・ERβ）を介したOH-PCBsの転写活性化能の分子機序について解析することを目的とした。
16種のOH-PCBsによるER アゴニスト転写活性化能を測定したところ、多くのOH-PCBsが活性を示し、異性体によって誘導倍率が大きく異なっていた。一方、一部の OH-PCBs は E2 と共処理時に相加・相乗作用を示した。そこでER転写活性化能に対するOH-PCBsの構造的特徴について理解するために、OH基置換位置の異なる3種のOH-CB30異性体の用量－応答関係を評価した。その結果、OH基置換位置がpara位 > meta位 > ortho位の順でEC50値が低く、高活性を示す傾向がみられた。次に、これらの物質におけるERリガンド結合ポケット内の結合状態を、in silicoシミュレーションで解析した。その結果、4’OH-CB30はE2と同様に、para位のOH基がE353と水素結合し、相互作用エネルギーは低値を示した。 一方で3’OH-CB30は、4’OH-CB30とは異なり、L346と水素結合を形成していたが、2’OH-CB30は近傍のアミノ酸残基と相互作用していなかった。これらのことから、OH基の置換位置の違いがERリガンドポケット内での相互作用に寄与し、転写活性化能に影響することが示唆された。
本研究の結果、多くのOH-PCBsは ERs に対しアゴニスト活性をもち、OH基や塩素の数・置換位置で異なる転写活性化能を有することが明らかとなった。

Research Products

• [Int'l Joint Research] Kyung Hee University(韓国)
  • Country Name: KOREA (REP. OF KOREA)
  • Counterpart Institution: Kyung Hee University
• [Journal Article] Organohalogen compounds in pet dog and cat: do pets biotransform natural brominated products in food to harmful hydroxlated substances? (2016)
  • Author(s): Mizukawa, H., Nomiyama, K., Nakatsu, S., Iwata, H., Yoo, J., Kubota, A., Yamamoto, M., Ishizuka, M., Ikenaka, Y., Nakayama, S. M. M., Kunisue, T. and Tanabe, S.
  • Journal Title: Environmental Science and Technology Volume: 50(1) Pages: 444-452
  • DOI: 10.1021/acs.est.5b04216
  • Peer Reviewed
• [Journal Article] In vitro and in silico evaluation of transactivation potencies of avian AHR1 and AHR2 by endogenous ligands: Implications for the physiological role of avian AHR2 (2016)
  • Author(s): Kim, I. S., Hwang, J. H., Hirano, M., Iwata, H. and Kim, E. Y.
  • Journal Title: Comparative Biochemistry and Physiology, Part C Volume: 187 Pages: 109
  • DOI: 10.1016/j.cbpc.2016.03.011
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Auto-induction mechanism of aryl hydrocarbon receptor 2 (AHR2) gene by TCDD-activated AHR1 AHR2 in the red seabream (Pagrus major) (2016)
  • Author(s): Bak, S. M., Iida, M., Soshilov, A. A., Denison, M. S., Iwata, H. and Kim, E. Y.
  • Journal Title: Archives of Toxicology Volume: 印刷中 Pages: 印刷中
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] HP genotyping of Vietnamese: global distribution of HPdel, complete deletion allele of the haptoglobin gene. (2015)
  • Author(s): Soejima, M., Agusa, T., Iwata, H., Fujihara, J., Kunito, T., Takeshita, H., Lan, V. .T., Minh, T. B., Takahashi , S., Trang, P. T. K., Viet, P. H., Tanabe, S. and Koda, Y.
  • Journal Title: Legal Medicine Volume: 17(1) Pages: 14-16
  • DOI: 10.1016/j.legalmed.2014.08.004
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] In vitro and in silico analyses for predicting hepatic cytochrome P450-dependent metabolic potencies of polychlorinated bipenyls in the baikal seal. (2015)
  • Author(s): Yoo, J., Hirano, M., Mizukawa, H., Nomiyama, K, Agusa, T., Kim, E. Y., Tanabe, S. and Iwata, H.
  • Journal Title: Environmental Science & Technology Volume: 49(24) Pages: 14588-14596
  • DOI: 10.1021/acs.est.5b03890
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Arsenic (＋3 Oxidation state) methyl transferase (AS3MT)遺伝子多型と無機ヒ素のメチル化能力との関連 (2015)
  • Author(s): 阿草哲郎，國頭恭，Nguyen Minh Tue， Vi Thi Mai Lan，Tu Binh Minh， Pham Thi Kim Trang，藤原純子，竹下治男，高橋真，Pham Hung Viet，田辺信介，岩田久人
  • Journal Title: 日本衛生学雑誌 Volume: 70 Pages: 186-196
  • DOI: 10.1265/jjh.70.186
  • Peer Reviewed
• [Presentation] Past, present, and future perspectives of environmental toxicology at Ehime University: advent of an era of the authentic animal study (2016)
  • Author(s): Iwata, H.
  • Organizer: International Symposium of Environmental Chemistry and toxicology - To Accelerate a Global Network of Environmental Researchers
  • Place of Presentation: 愛媛大学（愛媛県松山市）
  • Year and Date: 2016-03-19
  • Int'l Joint Research
• [Presentation] Assessment of effects of dioxins in the Baikal seal (Pusa Sibirica) by proteome and transcriptome analyses (2016)
  • Author(s): Lauan, M. C., Yoo, J., Agusa, T., Hirano, M., Kim, E. Y. and Iwata, H.
  • Organizer: International Symposium of Environmental Chemistry and toxicology - To Accelerate a Global Network of Environmental Researchers
  • Place of Presentation: 愛媛大学（愛媛県松山市）
  • Year and Date: 2016-03-19
  • Int'l Joint Research
• [Presentation] Effects of a naturally occurring dioxin, 1,3,7-tribromodibenzo-p-dioxin on the mysid transcriptome (2016)
  • Author(s): Hirano, M., Uchida, M., Kusano, T., Arizono, K. and Iwata, H.
  • Organizer: International Symposium of Environmental Chemistry and toxicology - To Accelerate a Global Network of Environmental Researchers
  • Place of Presentation: 愛媛大学（愛媛県松山市）
  • Year and Date: 2016-03-19
  • Int'l Joint Research
• [Presentation] Liver transcriptome analysis to assess the effects of prenatal exposure to bisphenol a of rat offspring (2016)
  • Author(s): Hoa, T. N., Yamamoto, K., Agusa, T., Hirano, M., Ochiai, M., Kim, E. Y. and Iwata, H.
  • Organizer: International Symposium of Environmental Chemistry and toxicology - To Accelerate a Global Network of Environmental Researchers
  • Place of Presentation: 愛媛大学（愛媛県松山市）
  • Year and Date: 2016-03-19
  • Int'l Joint Research
• [Presentation] Ecological factors drive natural selection pressure affecting avian dioxin receptor genotypes (2016)
  • Author(s): Hwang, J. H., Park, J. Y., Park, H. J., Bak, S. M., Hirano, M., Iwata, H., Park, Y. S. and Kim, E. Y.
  • Organizer: International Symposium of Environmental Chemistry and toxicology - To Accelerate a Global Network of Environmental Researchers
  • Place of Presentation: 愛媛大学（愛媛県松山市）
  • Year and Date: 2016-03-19
  • Int'l Joint Research
• [Presentation] Transcriptome analysis to identify adverse outcome pathways mediated by the aryl hydrocarbon receptor in TCDD-ｔreated red seabream embryos (2016)
  • Author(s): Iida, M., Fujii, S., Uhida, M., Nakamura, H., Kagami, Y., Agusa, T., Hirano, M., Bak, S. M., Kim, E. Y. and Iwata, H.
  • Organizer: International Symposium of Environmental Chemistry and toxicology - To Accelerate a Global Network of Environmental Researchers
  • Place of Presentation: 愛媛大学（愛媛県松山市）
  • Year and Date: 2016-03-19
  • Int'l Joint Research
• [Presentation] Effect of prenatal exposure of BPA on the transcriptome in chicken embryos (2016)
  • Author(s): Itoh, S., Agusa, T., Hirano, M., Ochiai, M., Kim, E. Y. and Iwata, H.
  • Organizer: International Symposium of Environmental Chemistry and toxicology - To Accelerate a Global Network of Environmental Researchers
  • Place of Presentation: 愛媛大学（愛媛県松山市）
  • Year and Date: 2016-03-19
  • Int'l Joint Research
• [Presentation] Hepatic transcriptome analysis to assess the effects of polychlorinated biphenyls on beagle dogs (2016)
  • Author(s): Tamura, S., Yoo, J., Mizukawa, H., Agusa, T., Hirano, M., Eguchi A., Nomiyama, K., Tanabe, S., Kim, E. Y. and Iwata, H.
  • Organizer: International Symposium of Environmental Chemistry and toxicology - To Accelerate a Global Network of Environmental Researchers
  • Place of Presentation: 愛媛大学（愛媛県松山市）
  • Year and Date: 2016-03-19
  • Int'l Joint Research
• [Presentation] Assessment of metabolic potential of polyhlorinated biphenyl congeners in the beagle dog liver by in vitro and in silico approaches (2016)
  • Author(s): Yoo, J., Hirano, M., Nomiyama, K., Kim, E. Y., Tanabe, S. and Iwata, H.
  • Organizer: International Symposium of Environmental Chemistry and toxicology - To Accelerate a Global Network of Environmental Researchers
  • Place of Presentation: 愛媛大学（愛媛県松山市）
  • Year and Date: 2016-03-19
  • Int'l Joint Research
• [Presentation] Structural preference of OH-PCBs to activate the Baikal Seal estrogen receptors (2016)
  • Author(s): Yoshinouchi, Y., Hirano, M., Nomiyama, K., Tanabe, S., Kim, E. Y. and Iwata, H.
  • Organizer: International Symposium of Environmental Chemistry and toxicology - To Accelerate a Global Network of Environmental Researchers
  • Place of Presentation: 愛媛大学（愛媛県松山市）
  • Year and Date: 2016-03-19
  • Int'l Joint Research
• [Presentation] OH-PCBｓによるバイカルアザラシ エストロゲン受容体α・β転写活性化能の評価 (2015)
  • Author(s): 芳之内結加，平野将司，野見山桂，田辺信介，金恩英，岩田久人
  • Organizer: 環境ホルモン学会第１８回研究発表会
  • Place of Presentation: 自治医科大学地域医療学情報センター（栃木県下野市）
  • Year and Date: 2015-12-11
• [Presentation] Effects of prenatal exposure to bisphenol A on the liver transcriptome of rat offspring (2015)
  • Author(s): Hoa, N. T., Yamamoto, K., Agusa, T., Hirano, M., Ochiai, M., Kim, E. Y. and Iwata, H.
  • Organizer: 環境ホルモン学会第１８回研究発表会
  • Place of Presentation: 自治医科大学地域医療学情報センター（栃木県下野市）
  • Year and Date: 2015-12-10
• [Presentation] In silico and in vitro screening of the effects of PPCPs on mysid ecdysteroid receptor signaling (2015)
  • Author(s): Hirano, M., Ohno, Y. and Iwata, H.
  • Organizer: 環境ホルモン学会第１８回研究発表会
  • Place of Presentation: 自治医科大学地域医療学情報センター（栃木県下野市）
  • Year and Date: 2015-12-10
• [Presentation] 鳥類の化学物質感受性を規定する分子機序 (2015)
  • Author(s): 岩田久人，平野将司， 金 恩英
  • Organizer: 第４２回日本毒性学会学術年会（The Japanese Society of Toxicology）
  • Place of Presentation: ホテル日航金沢（石川県金沢市）
  • Year and Date: 2015-06-30
  • Invited
• [Presentation] バイカルアザラシ エストロゲン受容体を介したビスフェノール類によるアンタゴニスト作用の評価 (2015)
  • Author(s): 芳之内結加、清水沙千子、李 鎮善、平野将司、鈴木賢一、中田晴彦、金 恩英、岩田久人
  • Organizer: 第24回環境化学討論会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2015-06-24
• [Presentation] Hepatic cytochrome P450-dependent metabolic potency of polychlorinated biphenyls in the baikal seal (Pula sibirica) (2015)
  • Author(s): Yoo, J., Hirano, M., Mizukawa, H., Nomiyama, K., Agusa, T., Kim, E. Y., Tanabe, S. and Iwata, H.
  • Organizer: 18th International Symposium on Pollutant Responses in Marine Organisms
  • Place of Presentation: クラリオンホテル＆コングレス（トロンハイム、ノルウェイ）
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Presentation] Identification of functional xenobiotic-responsive elements controlling AHR2 expression of red seabream (Pagrus major) (2015)
  • Author(s): Bak, S. M., Iida, M., Soshilov, A. A., Denison, M. S., Iwata, H. and Kim, E. Y.
  • Organizer: 18th International Symposium on Pollutant Responses in Marine Organisms
  • Place of Presentation: クラリオンホテル＆コングレス（トロンハイム、ノルウェイ）
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Presentation] Hepatic proteome analysis of baikal seals contaminated with dioxins and related compounds (2015)
  • Author(s): Lauan, M. C., Agusa, T., Hirano, M., Iwata, H., Kim, E. Y. and Yoo, J.
  • Organizer: 18th International Symposium on Pollutant Responses in Marine Organisms
  • Place of Presentation: クラリオンホテル＆コングレス（トロンハイム、ノルウェイ）
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Presentation] Comparison of hepatic cytochrome P450-dependent metabolic potencies of polychlorinated biphenyls in the caniformia, the baikal seal and the beagle dog. (2015)
  • Author(s): Yoo, J., Mizukawa, H., Nomiyama, K., Kim, E. Y., Tanabe, S. and Iwata, H.
  • Organizer: 18th International Symposium on Pollutant Responses in Marine Organisms
  • Place of Presentation: クラリオンホテル＆コングレス（トロンハイム、ノルウェイ）
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Presentation] In vitro and in silico assessment of transactivation potencies of baikal seal estrogen receptors (2015)
  • Author(s): Yoshinouchi, Y., Shimizu, S., Lee, J. S., Hirano, M., Agusa, T., Suzuki, K. T., Nakata, H., Kim, E. Y. and Iwata, H.
  • Organizer: 18th International Symposium on Pollutant Responses in Marine Organisms
  • Place of Presentation: クラリオンホテル＆コングレス（トロンハイム、ノルウェイ）
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Presentation] Ecological factors affecting natural selection of avian AHR1 genotypes (2015)
  • Author(s): Hwang, J. H., Park, H. J., Bak, S. M., Hirano, M., Iwata, H., Park, Y. S. and Kim, E. Y.
  • Organizer: 18th International Symposium on Pollutant Responses in Marine Organisms
  • Place of Presentation: クラリオンホテル＆コングレス（トロンハイム、ノルウェイ）
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Remarks] 岩田研究室のサイト日本語版
  • URL: http://ecotoxiwata.jp/
• [Remarks] 岩田研究室のサイト英語版
  • URL: http://ecotoxiwata.jp/en/index.html