ジーンターゲティングによるヒト発癌機構の解析

• Project/Area Number: 08267203
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Research Institution: Tohoku University (1997-1999); Japanese Foundation For Cancer Research (1996)
• Principal Investigator: 野田 哲生 東北大学, 大学院・医学系研究科, 教授 (10183550)
• Co-Investigator(Kenkyū-buntansha): 高野 洋志 東北大学, 大学院・医学系研究科, 助手 (00241555) ; 柴田 浩行 財団法人癌研究會, 癌研究所・細胞生物部, 研究員 (50260071) ; 芝 清隆 財団法人癌研究會, 癌研究所・細胞生物部, 主任研究員 (40196415)
• Project Period (FY): 1996 – 1999
• Project Status: Completed (Fiscal Year 1999)
• Budget Amount: ¥47,000,000 (Direct Cost: ¥47,000,000); Fiscal Year 1999: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 1998: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 1997: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 1996: ¥17,000,000 (Direct Cost: ¥17,000,000)
• Keywords: コンディショナル・ジーンターゲティング / APC遺伝子 / ケラチン14遺伝子 / βカテニン / WNTシグナル / ノックアウト・マウス / βカテニン遺伝子 / 大腸癌 / 膵癌 / コンディショナル・ノックアウト / アデノウイルスベクター / Cre組換え酵素 / がん抑制遺伝子 / ジーンターゲティング / モデルマウス / VHL遺伝子 / MLH1遺伝子 / p53遺伝子

Research Abstract

我々は、世界に先駆けて、Cre組換え酵素遺伝子を発現する組換えアデノウイルスを用いた、新たなコンディショナル・ジーンターゲティング法を開発し、これを用いることによりAPC遺伝子の不活化が(1)マウスの胆道上皮を癌化させること、及び(2)マウスの膵臓上皮を癌化させるには、さらにp53遺伝子が不活化されている必要があることを明かにした。この膵癌ではβカテニンが細胞質及び核内に貯留しており、APC遺伝子の不活化がWNTシグナルの活性化を介して細胞を癌化させたことが示唆されたが、このβカテニンの貯留の頻度及び程度は、p53変異の有無との関係は認められず、p53遺伝子変異はWNTシグナルが活性された細胞が増殖を始めるために必要であると推察された。一方、我々はマウスの発生における特定の細胞系列でのみCreを発現するトランスジェニックマウスを作成し、これをコンディショナルノックアウトマウスと交配させることにより、マウスの個体の発生におけるAPC遺伝子の機能の解析を行っている。本年度は、ケラチン14遺伝子のプロモーターを用い、表皮の基底細胞特異的にCreを発現するマウスを樹立し、これを交配することにより表皮でのみAPC遺伝子が不活化されているマウスを作成した。この結果、表皮でもβカテニンの貯留が観察され、このWNTシグナルの活性化された基底細胞では細胞増殖の刺激も観察されたが、同時に著明なアポトーシスが観察された。

Research Products

• [Publications] Noda, T., et al.: "Renal Carcinogenesis, hemangiomatosis, and embryonic lethality caused by a gern-line Tsc2 mutation in mice"Cancer. Res.. 59. 1206-1211 (1999)
• [Publications] Noda, T., et al.: "RhoN, a novel small GTP-binding protein expressed predominantly in neurons and hepatic stellate cells"Mol. Brain. Res.. 67. 74-81 (1999)
• [Publications] Noda, T., et al.: "Fas ligand-mediated exocrinopathy rersembling Sjogren's syndrome in mice transgenic for IL-10"J. Immunol.. 162. 2488-2494 (1999)
• [Publications] Noda, T., et al.: "α-Catenin deficient F9 cells differenciate into signet ring cells"Am. J. Pathol.. 154(5). 1323-1327 (1999)
• [Publications] Noda, T., et al.: "Altered cochlear fibrocytes in a mous model of DFN3 nonsyndromic deafness"Science. 285. 1408-1411 (1999)
• [Publications] Noda, T., et al.: "Disruption of cholecystokinin (CCK)-B receptor gene did not modify bile or pancreatic secretion or pancreatic growth : A study in CCK-B receptor gene knockout mice"Pancreas. 19(2). 114-118 (1999)
• [Publications] Noda, T., et al.: "Abnormal features in skeletal muscle from mice lacking mitsugumin29"J. Cell Biol.. 147(7). 1473-1480 (1999)
• [Publications] Noda, T., et al.: "Impairment of anrigen-presenting cell function in mice lacking expression of OX40 ligand"J. Exp. Med.. 191(2). 365-374 (2000)
• [Publications] Noda, T., et al.: "Mmh/Ogg1 gene inactivation resulfs in accumulation of 8-OH-G in Mice"Proc. Natl. Acad. Sci. USA. ( in press ). (2000)
• [Publications] Noda, T., et al.: "Targeted disruption of the homeobox transcription factor Bapxl results in lethal skeletal dysplasia eith asplenia and gastroduodenal malformation"Gene to Cells. ( in press ). (2000)
• [Publications] Noda, T., et al.: "Anal. Biochem."successful germ-line transmission of chimeras generated by coculture aggregation with J1 ES cells and eight-cell embryos ( in press ). (1999)
• [Publications] Noda, T., et al.: "Proceedings"Underdeveloped uterus and reduced estrogen responsiveness in mice with disruption of the estrogen-responsive finger protein (efp) gene, which is a direct of estrogen receptor α(ER α) ( in press ). (1999)
• [Publications] Y.Yonekawa,T.Noda et al.: "Defect in synaptic vesicle precursor transport and neuronal cell death in DIF1A motor protein-deficient mice." J.Cell Biol.141. 431-441 (1998)
• [Publications] M.Saitou,T.Noda et al.: "Occuludin-deficient embryonic stem cells can differentiate into polarized epithelial cells bearing tight junctions." J.Cell Biol.141(2). 397-408 (1998)
• [Publications] H.Kawate,T.Noda et al.: "Separation of Killing and tumorigenic effects of an alkylating agent in mice defective in two of the DNA repair genes." Proc.Natl.Acad.Sci.USA.95. 5116-5120 (1998)
• [Publications] H.Takeshima,T.Noda et al.: "Embryonic lethality and abnormal cardiac myocytes mice lacking ryanodine receptor type2." EMBO J.17. 3309-3316 (1998)
• [Publications] T.Manabe,T.Noda et al.: "Facilitation of long-term potentiation and memory in mice lacking nociceptin receptors." Nature. 394. 577-581 (1998)
• [Publications] S.Hiratsuka,T.Noda et al.: "Fit-1 lacking the tyrosine kinase domain is sufficient for normal development and angiogenesis in mice." Proc.Natl.Acad.Sci.95. 9349-9354 (1998)
• [Publications] M.Nishi, T.Noda et al.: "Unrestrained nociceptive response and disregulation of hearing ability in mice lacking noniceptin/orphanin FQ receptor." EMBOJ.16. 1854-1864 (1997)
• [Publications] H.Suzuki, T.Noda et al.: "A role for macrophage scavenger receptors in atherosclerosis and susceptivility to infection." Nature. 386. 292-296 (1997)
• [Publications] K.Shida, T.Noda et al.: "Creation of libraries with long open reading frames by polymerization of a microgene." Proc.Natl.Acad.Sci.USA. 94. 3805-3810 (1997)
• [Publications] M.Niki, T.Noda et al.: "Hematopoiesis in the fetal liver is impaired by targeted mutagenesis of a gene encoding a non-DNA binding subunit of the transcription factor,PEBP2/CBP." Proc.Natl.Acad.Sci USA. 94. 5697-5702 (1997)
• [Publications] A.Terunuma, T.Noda et al.: "A novel genetic system to isolate a domonant negative effecter on DNA-binding activity of Oct-2" Nucl.Acid.Res.,. 25. 1984-1990 (1997)
• [Publications] Y.Wakabayashi, T.Noda et al.: "Genetic and physical delineation of the region of the mouse deafness mutation shaker-2." Biochem.Biophis.Res.Comm.232. 107-110 (1997)
• [Publications] Y.Takei, T.Noda et al.: "Delayed development of nervous system in mice homozygous for disrupted microtubuleassociated protein IB (MAP1B) gene." J.Cell Biol.137. 1615-1626 (1997)
• [Publications] K.Shiba, T.Noda et al.: "Human lysyl-tRNA synthetase accepts N73 variants and rescues E.coli double-defective mutant." J.Biol.Chem.272. 22809-22816 (1997)
• [Publications] H.Shibata, T.Noda et al.: "Rapid colorectal adenoma formation initiated by conditional targeting of Apc gene" Science. 278. 120-123 (1997)
• [Publications] J.Aruga, T.Noda et al.: "Mouse Zicl is involved in cerebellar development." J.Neurosci. (in press). (1998)
• [Publications] J.Toshima, T.Noda et al.: "Structural organization and chromosomal localization of the mouse Tesk1 (testis-specific protein kinase 1) gene." Gene. (in press). (1998)
• [Publications] C.Ikebe, T.Noda et al.: "Mouse LIM-kinase 2 gene : cDNA cloning,genomic organization and tissue-specific expression of two alternatively initiated transcripts." Genomics. (in press). (1998)
• [Publications] Matsumoto,M.,Noda,T.,et al.: "Ataxia and epileptic seizure in mice lacking type 1 inositol 1,4,5-trisphophate receptor." Nature. 379. 168-171 (1996)
• [Publications] Sakaguchi,S.Noda,T.,et al.: "Loss of cerebellar Purkinje cells in aged mice homozygous for a disrupted PrP gene." Nature. 380. 528-531 (1996)
• [Publications] Takeshima,H.,Noda,T.,et al.: "Generation and characterization of mutant mice lacking ryanodine receptor type3." J.Biol.Chem.271. 19649-19652 (1996)
• [Publications] Nagata,A.,Noda,T.,et al.: "Gprotein-coupled cholecystokinin-B/gastrin receptors are responsible for physiological cell growth of the stomach mucosa in vivo." Proc.Natl.Acad.Sci.USA. 93. 11825-11830 (1996)
• [Publications] Nishi,M.,Noda,T.,et al.: "Unrestrained nociceptive response and disregulation of hearing ability in mice lacking noniceptin/orphaninFQ receptor." EMBOJ.(in press). (1997)
• [Publications] Suzuki,H.,Noda,T.,et al.: "A role for macrophage scavenger receptors in atherosclerosis and susceptibility to infection." Nature. (in press). (1997)
• [Publications] Shiba,K.,Noda,T.,et al.: "Creation of libraries with long open reading frames by polymerization of a microgene." Proc.Natl.Acad.Sci.USA. (in press). (1997)
• [Publications] Niki,M.,Noda,T.,et al.: "Hematopoiesis in the fetal liver is impaired by targeted mutagenesis of a gene encoding a non-DNA binding subunit of the transcription factor,PEBP2/CBP." Proc.Natl.Acad.Sci.USA. (in press). (1997)