2001 Fiscal Year Annual Research Report

遺伝子改変マウスを用いた環境異物モニタリングモデル動物の開発

Research Project

Project/Area Number	12794009
Research Institution	UNIVERSITY OF TSUKUBA
Principal Investigator	山本雅之筑波大学, 基礎医学系, 教授 (50166823)
Co-Investigator(Kenkyū-buntansha)	原田孝則残留農薬研究所, 毒性第2部, 部長青木康展国立環境研究所, 環境研究部, 室長 (20159297) 高橋智筑波大学, 基礎医学系, 教授 (50271896)
Keywords	異物代謝 / 環境異物 / 転写制御 / モデル動物 / bZip型転写因子 / bHLH-PAS型転写因子
Research Abstract	今年度は以下の成果を得た. (1)keap1欠失マウスとkeap7::nrf2複合欠失マウスの作製培養細胞を用いた実験から,Keap1はNrf2の活性を負に制御する因子であることが予測された.個体におけるKeap1の機能を明らかにするため,keap1欠失マウスを作製し,その表現型を解析したところ,同マウスの肝臓では,Nrf2の恒常的な活性化が起こっていることが明らかになった.さらに,個体におけるNrf2とKeap1の相互関係を明らかにするためにkeap1::nrf複合欠失マウスを作製し,肝臓におけるNrf2の標的遺伝子の発現を検討したところ,Nrf2の恒常的活性化は解除されていた.生体において,実際にKeap1がNrf2の抑制因子として機能していることが証明された. (2)ヒト型AhRマウスの作製ヒト型AhRは,マウス(C57/Black6)型AhRに比べて,ダイオキシンに対する親和性が著しく低い.ダイオキシンに対する感受性がヒトと同程度のマウスを得るために,マウスAhR遺伝子座をヒト型AhR cDNAで置換したマウスを作製した.同マウスの胎児に対するダイオキシンの影響を調べたところ,野生型マウス(C57/Black6)の胎児では,口蓋裂と重度の水腎症が全例で認められたのに対して,ヒト型AhRマウスの胎児では,口蓋裂は全く認められず,軽度の水腎症が低い頻度で認められるのみであった.AhRのダイオキシンに対する親和性が,ダイオキシンに対する個体の反応性を決定していることが確認された. (3)Nrf2欠損マウスの環境異物・薬剤にたいする応答反応の検討 Nrf2欠失マウスでは,異物代謝系第2相酵素群の誘導が欠落していることから,様々な外来の化学物質により障害を受けやすいことが予測された.解熱鎮痛消炎剤として頻用されているアセトアミノフェンを投与したところ,nrf2欠失マウスでは急性肝障害が高頻度に誘発された.また,ディーゼルエンジンの排気ガスに曝露したところ,nrf2欠失マウスではDNAの塩基の修飾(DNAアダクト)が高頻度に生成されていることが明らかになった.Nrf2が,実際に生体において,外来の化学物質の代謝に重要な役割を果たしていることが証明された.

Research Products

(31 results)

All Other

All Publications (31 results)

[Publications] Enomoto, A., et al.: "High sensitivity of Nrf2 knockout mice to acetoaminophen hepatotoxicity associated with decreased expression of ARE-regulated drug metabolizing enzyme and antioxidant genes"Toxicol. Sci.. 59. 169-177 (2001)
[Publications] Tabata, M. et al.: "Stimulation of GATA-2 as a mechanism of hydrogen peroxide suppression in hypoxia-induced erythropoietin gene expression"J. Cell. Physiol.. 186. 260-267 (2001)
[Publications] Ramos-Gomez, M.: "Sensitivity to carcinogenesis is increased and chemoprotective efficacy of enzyme inducers is lost in nrf2 transcription factor-deficient mice"Proc. Natl. Acad. Sci. USA. 98. 3410-3415 (2001)
[Publications] Kwak, M-K., et al.: "Role of Transcription factor Nrf2 in the induction of hepatic phase 2 and antioxidative enzymes in vivo by the cancer chemopreventive agent. 3H-1, 2-dithiole-3-thione"Mol. Med.. 7. 135-145 (2001)
[Publications] Kanezaki, R., et al.: "Transcription factor BACHI is recruited to nucleus by its novel alternative spliced isoform expressed in testis"J. Biol. Chem.. 276. 7278-7284 (2001)
[Publications] Kobayashi, M., et al.: "The homeobox protein Six3 interacts with the Groucho corepressor and acts as a transcriptional repressor in eye and forebrain formation"Dev. Biol.. 232. 315-326 (2001)
[Publications] Hosoya, T. et al.: "Defective development of secretary neurons in the hypothalamus of Arnt2-knockout mice"Genes Cells. 6. 361-374 (2001)
[Publications] Umetani, M.: "Function of GATA transcription factors in the induction of endothelial VCAM-1 by TNF-α"Arterioscler. Thromb. Vasc. Biol.. 21. 917-922 (2001)
[Publications] Kobayashi, M. et al.: "Positive regulation by GATA-1 of GATA-1 gene hematopoietic regulatory domain in zebrafish embryos in vivo"Development. 128. 2341-2350 (2001)
[Publications] Kim, Y-C. et al.: "Hemin-induced activation of the Thioredoxin gene by Nrf2 : a differential regulation of the antioxidant responsive element (ARE) by switch of its binding factors"J. Biol. Chem.. 276. 18399-18406 (2001)
[Publications] Ogawa, K.: "Heme mediates de-repression of Maf recognition element through direct binding to transcription factor Bach1"EMBO J.. 20. 2835-2843 (2001)
[Publications] Yoh, K.: "Nrf2 deficient mouse : a novel model for lupus nephritis"Kidney Int.. 60. 1343-1353 (2001)
[Publications] Katoh, Y. et al.: "Two domains of Nrf2 cooperatively bind CBP (CREB binding protein) and synergistically activate transcription"Genes Cells. 6. 857-868 (2001)
[Publications] Shimizu, R. et al.: "In vivo requirements for functional GATA-1 domains during primitive and definitive erythropoiesis"EMBO J.. 20. 5250-5260 (2001)
[Publications] Sun, J.et al.: "The promoter of mouse transcription repressor bach 1 is regulated by Sp 1 and transactivated by Bach1"J. Biochem.. 130. 385-392 (2001)
[Publications] McMahon, M. et al.: "The Cap' n' Collar basic leucine zipper transcription factor Nrt2 controls both constitutive and inducible expression of intestinal detoxification and glutathione biosynthetic enzymes"Cancer Res.. 61. 3299-3307 (2001)
[Publications] Imagawa, S. et al.: "Levels of vascular endothelial growth factor are elevated in patients with obstructive sleep apnea-hypopnea syndrome"Blood. 98. 1255-1257 (2001)
[Publications] Kwak, M.K.et al.: "Role of phase 2 enzyme induction in chemoprotection by dithiolethiones"Mutat Res. 481. 305-315 (2001)
[Publications] Yoh, K.et al.: "Transgenic overexpression Of MafK suppresses T cell proliferation and function in vivo"Genes Cells. 6. 1055-1066 (2001)
[Publications] Fujimoto, T. et al.: "Stepwise divergence of primitive and definitive hematopoietic and endothelial cell lineages"Genes Cells. 6. 1113-1127 (2001)
[Publications] Aoki, Y. et al.: "Accelerated DNA adducts formation in the lung of the Nrf2 knockout mouse exposed to diesel exhaust"Toxicol. Appl. Pharmacol. 173. 154-160 (2001)
[Publications] Cho, H.Y.et al.: "Linkage analysis of susceptibility to hyperoxic lung injury : role of Nrf2 as a candidate gene"Am. J. Res. Cell Mol. Biol.. (in press).
[Publications] Morimitsu, Y: "A sulforaphane analogue that potentially activates the Nrf2-dependent detoxification pathway"J. Biol. Chem.. (in press).
[Publications] Kirito, K. et al.: "A functional role of Stat3 in in vivo megakaryopoiesis"Blood. (in press).
[Publications] Kusunoki, H. et al.: "Solution structure of the DNA-binding domain of MafG"Nature Str. Biol.. (in press).
[Publications] Kwak, M.K.et al.: "Regulation of the expression of Nrt2 transcription factor by cancer chemopreventive agents through antioxidant response element (ARE)-like sequences in its proximal promoter region"Mel. Cell. Biol.. (in press).
[Publications] Ueno, Y. et al.: "Elimination of Pasteurella pneumotropica from a Contaminated Mouse Colony by Oral Administration of Enrofloxacin"Exp. Animals. (in press).
[Publications] 山本雅之編: "「遺伝子の構造と機能」シリーズ・バイオサイエンスの新世紀"日本生化学会編集,共立出版. 178 (2001)
[Publications] 勝岡史城, 山本雅之: "新臨床医のための分子医学シリーズ/造血幹細胞研究のいまと臨床応用「遺伝子操作マウスを用いた血液疾患モデルの作製」"羊土社. (2001)
[Publications] 中島修, 山本雅之: "Annual Review 血液 2002 「X染色体連鎖型鉄芽球性貧血モデルおよび鉄代謝異常関連遺伝子改変動物の分子生物学的解析」"中外医学社(印刷中).
[Publications] 鈴木教朗, 山本雅之: "造血幹細胞:基礎から遺伝子治療・再生医療へ「造血幹細胞の転写調節機構」"中外医学社(印刷中).

2001 Fiscal Year Annual Research Report

遺伝子改変マウスを用いた環境異物モニタリングモデル動物の開発

Principal Investigator

山本 雅之 筑波大学, 基礎医学系, 教授 (50166823)

Research Products

[Publications] Enomoto, A., et al.: "High sensitivity of Nrf2 knockout mice to acetoaminophen hepatotoxicity associated with decreased expression of ARE-regulated drug metabolizing enzyme and antioxidant genes"Toxicol. Sci.. 59. 169-177 (2001)

[Publications] Tabata, M. et al.: "Stimulation of GATA-2 as a mechanism of hydrogen peroxide suppression in hypoxia-induced erythropoietin gene expression"J. Cell. Physiol.. 186. 260-267 (2001)

[Publications] Ramos-Gomez, M.: "Sensitivity to carcinogenesis is increased and chemoprotective efficacy of enzyme inducers is lost in nrf2 transcription factor-deficient mice"Proc. Natl. Acad. Sci. USA. 98. 3410-3415 (2001)

[Publications] Kwak, M-K., et al.: "Role of Transcription factor Nrf2 in the induction of hepatic phase 2 and antioxidative enzymes in vivo by the cancer chemopreventive agent. 3H-1, 2-dithiole-3-thione"Mol. Med.. 7. 135-145 (2001)

[Publications] Kanezaki, R., et al.: "Transcription factor BACHI is recruited to nucleus by its novel alternative spliced isoform expressed in testis"J. Biol. Chem.. 276. 7278-7284 (2001)

[Publications] Kobayashi, M., et al.: "The homeobox protein Six3 interacts with the Groucho corepressor and acts as a transcriptional repressor in eye and forebrain formation"Dev. Biol.. 232. 315-326 (2001)

[Publications] Hosoya, T. et al.: "Defective development of secretary neurons in the hypothalamus of Arnt2-knockout mice"Genes Cells. 6. 361-374 (2001)

[Publications] Umetani, M.: "Function of GATA transcription factors in the induction of endothelial VCAM-1 by TNF-α"Arterioscler. Thromb. Vasc. Biol.. 21. 917-922 (2001)

[Publications] Kobayashi, M. et al.: "Positive regulation by GATA-1 of GATA-1 gene hematopoietic regulatory domain in zebrafish embryos in vivo"Development. 128. 2341-2350 (2001)

[Publications] Kim, Y-C. et al.: "Hemin-induced activation of the Thioredoxin gene by Nrf2 : a differential regulation of the antioxidant responsive element (ARE) by switch of its binding factors"J. Biol. Chem.. 276. 18399-18406 (2001)

[Publications] Ogawa, K.: "Heme mediates de-repression of Maf recognition element through direct binding to transcription factor Bach1"EMBO J.. 20. 2835-2843 (2001)

[Publications] Yoh, K.: "Nrf2 deficient mouse : a novel model for lupus nephritis"Kidney Int.. 60. 1343-1353 (2001)

[Publications] Katoh, Y. et al.: "Two domains of Nrf2 cooperatively bind CBP (CREB binding protein) and synergistically activate transcription"Genes Cells. 6. 857-868 (2001)

[Publications] Shimizu, R. et al.: "In vivo requirements for functional GATA-1 domains during primitive and definitive erythropoiesis"EMBO J.. 20. 5250-5260 (2001)

[Publications] Sun, J.et al.: "The promoter of mouse transcription repressor bach 1 is regulated by Sp 1 and transactivated by Bach1"J. Biochem.. 130. 385-392 (2001)

[Publications] McMahon, M. et al.: "The Cap' n' Collar basic leucine zipper transcription factor Nrt2 controls both constitutive and inducible expression of intestinal detoxification and glutathione biosynthetic enzymes"Cancer Res.. 61. 3299-3307 (2001)

[Publications] Imagawa, S. et al.: "Levels of vascular endothelial growth factor are elevated in patients with obstructive sleep apnea-hypopnea syndrome"Blood. 98. 1255-1257 (2001)

[Publications] Kwak, M.K.et al.: "Role of phase 2 enzyme induction in chemoprotection by dithiolethiones"Mutat Res. 481. 305-315 (2001)

[Publications] Yoh, K.et al.: "Transgenic overexpression Of MafK suppresses T cell proliferation and function in vivo"Genes Cells. 6. 1055-1066 (2001)

[Publications] Fujimoto, T. et al.: "Stepwise divergence of primitive and definitive hematopoietic and endothelial cell lineages"Genes Cells. 6. 1113-1127 (2001)

[Publications] Aoki, Y. et al.: "Accelerated DNA adducts formation in the lung of the Nrf2 knockout mouse exposed to diesel exhaust"Toxicol. Appl. Pharmacol. 173. 154-160 (2001)

[Publications] Cho, H.Y.et al.: "Linkage analysis of susceptibility to hyperoxic lung injury : role of Nrf2 as a candidate gene"Am. J. Res. Cell Mol. Biol.. (in press).

[Publications] Morimitsu, Y: "A sulforaphane analogue that potentially activates the Nrf2-dependent detoxification pathway"J. Biol. Chem.. (in press).

[Publications] Kirito, K. et al.: "A functional role of Stat3 in in vivo megakaryopoiesis"Blood. (in press).

[Publications] Kusunoki, H. et al.: "Solution structure of the DNA-binding domain of MafG"Nature Str. Biol.. (in press).

[Publications] Kwak, M.K.et al.: "Regulation of the expression of Nrt2 transcription factor by cancer chemopreventive agents through antioxidant response element (ARE)-like sequences in its proximal promoter region"Mel. Cell. Biol.. (in press).

[Publications] Ueno, Y. et al.: "Elimination of Pasteurella pneumotropica from a Contaminated Mouse Colony by Oral Administration of Enrofloxacin"Exp. Animals. (in press).

[Publications] 山本雅之 編: "「遺伝子の構造と機能」シリーズ・バイオサイエンスの新世紀"日本生化学会編集,共立出版. 178 (2001)

[Publications] 勝岡史城, 山本雅之: "新臨床医のための分子医学シリーズ/造血幹細胞研究のいまと臨床応用「遺伝子操作マウスを用いた血液疾患モデルの作製」"羊土社. (2001)

[Publications] 中島修, 山本雅之: "Annual Review 血液 2002 「X染色体連鎖型鉄芽球性貧血モデルおよび鉄代謝異常関連遺伝子改変動物の分子生物学的解析」"中外医学社(印刷中).

[Publications] 鈴木教朗, 山本雅之: "造血幹細胞:基礎から遺伝子治療・再生医療へ 「造血幹細胞の転写調節機構」"中外医学社(印刷中).

山本雅之筑波大学, 基礎医学系, 教授 (50166823)

[Publications] 山本雅之編: "「遺伝子の構造と機能」シリーズ・バイオサイエンスの新世紀"日本生化学会編集,共立出版. 178 (2001)

[Publications] 鈴木教朗, 山本雅之: "造血幹細胞:基礎から遺伝子治療・再生医療へ「造血幹細胞の転写調節機構」"中外医学社(印刷中).