造血器腫瘍の遺伝子診断法の開発と実用化

• Project/Area Number: 06283206
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Research Institution: The University of Tokyo
• Principal Investigator: 平井 久丸 東京大学, 医学部・附属病院, 助教授 (90181130)
• Co-Investigator(Kenkyū-buntansha): 今井 陽一 東京大学, 医学部・附属病院, 医員 ; 高橋 宗春 東京大学, 医学部・附属病院, 助手 ; 千葉 滋 東京大学, 医学部・附属病院, 助手 (60212049) ; 山形 哲也 東京大学, 医学部・附属病院, 医員 ; 黒川 峰夫 東京大学, 医学部・附属病院, 医員 ; 小川 誠司 東京大学, 医学部・附属病院, 医員 ; 平野 直人 東京大学, 医学部(病), 医員 ; 田中 智之 東京大学, 医学部(病), 助手 (50227154)
• Project Period (FY): 1994 – 1999
• Project Status: Completed (Fiscal Year 1999)
• Budget Amount: ¥109,000,000 (Direct Cost: ¥109,000,000); Fiscal Year 1999: ¥25,000,000 (Direct Cost: ¥25,000,000); Fiscal Year 1998: ¥22,000,000 (Direct Cost: ¥22,000,000); Fiscal Year 1997: ¥20,000,000 (Direct Cost: ¥20,000,000); Fiscal Year 1996: ¥18,000,000 (Direct Cost: ¥18,000,000); Fiscal Year 1995: ¥12,000,000 (Direct Cost: ¥12,000,000); Fiscal Year 1994: ¥12,000,000 (Direct Cost: ¥12,000,000)
• Keywords: 白血病 / Evi-1 / 核内蛋白質 / zinc finger / MAP kinase / JNK / ストレス刺激 / アポトーシス / 急性リンパ性白血病 / 非ホジキンリンパ腫 / 染色体欠失 / 癌抑制遺伝子 / 発癌機構 / FISH法 / t(3;21)染色体転座 / AML1 / EVI-1キメラ遺伝子 / 転写因子 / リン酸化 / レドックス制御 / 造血器腫瘍 / 染色体転座 / 11q23転座 / MLL遺伝子 / MEN遺伝子 / 転写伸長活性 / AML1遺伝子 / 慢性骨髄性白血病 / Evi-1融合蛋白質 / 転写活性 / 造血細胞 / 細胞増殖 / 細胞分化 / キメラ蛋白質 / 転写制御因子 / 癌抑制 / p16遺伝子 / ALL

Research Abstract

Evi-1は2つのcluster(第1 zinc finger領域、第2 zinc finger領域)に分かれた10個のzinc finger構造を持つ核内蛋白質をコードし、ヒトでは染色体の3q26上に存在する。Evi-1は正常骨髄細胞ではほとんど発現が認められないが、ヒトの白血病において高発現をすることが知られている。これらのことから、Evi-1は造血細胞の悪性化に深く関与する遺伝子であり、その機能を明らかにすることは造血細胞の悪性化の機構を解明する上で重要である。本研究ではヒトの白血病発症に深く関わるEvi-1遺伝子産物の機能についてさらに解析を進め、以下のことを明らかとした。Evi-1はTGFβの細胞内シグナル伝達分子の一つであるSmad3と核内で結合してTGFβシグナルを阻害するが、この機序には、転写抑制因子であるCtBPがヒストン脱アセチル化酵素(HDAC1)をリクルートしてEvi-1に会合する機構が関与することが明らかとなった。また、Evi-1は、種々のストレスシグナルによるJNKの活性化を特異的に抑制することを明らかにした。Evi-1はJNKと結合し、この結合がEvi-1によるJNKの抑制に必須であった。Evi-1は、JNKによるFasLの発現誘導や細胞のアポトーシスの誘導を抑制した。これらの結果から、Evi-1はJNKと結合し、その機能を不活化することによってアポトーシスを抑制し、細胞の悪性化に関わることが明らかになった。このようにEvi-1は種々のメカニズムを通して細胞の増殖制御を破綻させ、造血細胞を悪性化に導くと考えられる。

Research Products

• [Publications] Maki K: "Transcriptional inhibition of p53 by the MLL/MEN chimeric protein found in myeloid leukemia"Blood. 93. 3216-3224 (1999)
• [Publications] Honda H: "Expression of E2A-HLF chimeric protein induced T-cell apoptosis, B-cell maturation arrest and development of acute lymphoblastic leukemia"Blood. 93. 2780-2790 (1999)
• [Publications] Hirai H: "Molecular characterization of the genomic breakpoints in a case of t(3;21)(q26;q22)"Genes chromosomes Cancer. 26. 92-96 (1999)
• [Publications] Shimizu K: "Mouse Jagged physically interacts with Notch2 and other Notch receptors: assessment by quantitative methods"J. Biol. Chem.. 274. 32961-32969 (1999)
• [Publications] Honda H: "Acquired loss of p53 induced blastic transformation of p210bcr/abl-expressing hematopoietic cells: A mouse model for blastic crisis of human CML"Blood. (In press).
• [Publications] Nakamoto T: "CIZ: a zinc finger protein that interacts with p130cas and activates the expression of matrix metalloproteinases"Mol. Cell. Biol.. (In press).
• [Publications] Kurokawa M: "The oncoprotein Evi-1 represses TGFβ signalling by inhibiting Smad3." Nature. 394. 92-96 (1998)
• [Publications] Honda H: "Development of acute lymphoblastic leukemia and myeloproliferative disorder in transgenic mice expressing p210bcr/abl : a novel transgenic model for human Ph1-positive leukemias." Blood. 91. 2067-2075 (1998)
• [Publications] Tanaka K: "The AML1/ETO(MTG8) and AML1/Evi-1 leukemia-associated chimeric oncoproteins accumulate PEBP2β(CBFβ) in the nucleus more efficiently than wild-type AML1." Blood. 91. 1688-1699 (1998)
• [Publications] Kanda Y: "Overexpression of the MEN/ELL protein,an RNA polymerase II elongation factor,results in transformation of Rat1 cells with dependence on the lysine-rich region." J.Biol.Chem.273. 5248-5252 (1998)
• [Publications] Kurokawa M: "The t(3;21) fusion product,AML1/Evi-1,interacts with Smad3 and blocks TGFβ-mediated growth inhibition of myeloid cells." Blood. 92. 4003-4012 (1998)
• [Publications] Honda H: "Cardiovascular anomaly,impaired actin bundling and resistance to Src-induced transformation in mice lacking p130Cas." Nature Genet. 19. 361-365 (1998)
• [Publications] Mitani K: "Generation of the AML1/Evi-1 fusion gene in the t(3;21)(q26;q22)causes blastic crisis in chronic myelocytic leukemia." EMBO J.13. 504-510 (1994)
• [Publications] Tanaka T: "Evi-1 raises AP-1 activity and stimulates c-fos promoter transactivation with dependence on the second zinc finger domain." J.Biol.Chem.269. 24020-24026 (1994)
• [Publications] Tanaka T: "An acute myeloid leukemia gene,AML1,regulates hemopoietic myeloid cell differentiation and transcriptional activation antagonistically by two alternative spliced forms." EMBO J.14. 341-350 (1995)
• [Publications] Tanaka T: "Dual functions of the AML1/Evi-1 chimeric protein in the mechanism of leukemogenesis in t(3;21)leukemias." Mol.Cell.Biol.15. 2383-2392 (1995)
• [Publications] Tanaka T: "The extracellular signal-regulated kinase pathway phosphorylates AML1,an acute myeloid leukemia gene product,and potentially regulates its transactivation ability." Mol.Cell.Biol.16. 3967-3979 (1996)
• [Publications] Kurokawa M: "A conserved cysteine residue in the runt homology domain of AML1 is required for the DNA-binding activity and the transforming activity on fibroblasts." J.Biol.Chem.271. 16870-16876 (1996)
• [Publications] Kurokawa M.: "Overexpression of the human AML1b proto-oncoprotein leads to neoplastic transformation of NIH3T3 cells." Oncogene.12. 883-892 (1996)
• [Publications] Yamagata T: "Molecular cloning of a novel IRF family transcription factor,ICSAT/Pip/LSIRF,that negatively regulates the activity of the interferon-regulated genes." Mol.Cell.Biol.16. 1283-1294 (1996)
• [Publications] Nakamoto T.: "Direct binding of C-terminal region of p130Cas to SH2 and SH3 domains of Src kinase." J.Biol.Chem.271. 8959-8965 (1996)
• [Publications] Ogawa S: "Structurally altered Evi-1 transcript is generated by inv(3)(q21q26) in a cell line derived from chronic myelocytic leukemias in blastic crisis." Oncogene. 13. 183-192 (1996)
• [Publications] Tanaka T.: "The extracellular signal-regulated kinase pathway phosphorylates AML1,an acute myeloid leukemia gene product,and potentially regulates its transactivation ability." Mol.Cell.Biol.16. 3967-3979 (1996)
• [Publications] Kurokawa M: "A conserved cysteine residue in the runt homology domain of AML1 is required for the DNA-binding activity and the transforming activity on fibroblasts." J.Biol.Chem.271. 16870-16876 (1996)
• [Publications] Tanaka T: "An acute myeloid leukemia gene, AML1, regulates hemopoietic myeloid cell differentiation and transcriptional activation antagonistically by two alternative spliced forms." EMBOJ. 14. 341-350 (1995)
• [Publications] Tanaka T: "Dual functions of the AML1/Evi-1 chimeric protein in the mechanism of leukemogenesis in t(3;21)leukemias." Mol.Cell.Biol.15. 2383-2392 (1995)
• [Publications] Mitani K: "Growth inhibition of leukemic cells carrying the t(3;21)by the AML1/EVI-1 specific antisense oligonucleotide." Brit.J.Hematol.90. 711-714 (1995)
• [Publications] Miyoshi H: "Alternative splicing and genomic structure of the AML1 gene involved acute myeloid leukemia." Nucleic A cid Res.23. 2762-2769 (1995)
• [Publications] Tanaka K: "Increased expression of AML1 during retinoic acid-induced differentiation of U937 cells." Biophys.R es.Commun.211. 1023-1030 (1995)
• [Publications] Kurokawa M: "The AML1/Evi-1 fusion protein generated in the t(3;21)translocation exhibits transforming activity on Rat1 fibroblasts with dependence on the Evi-1 sequence." Oncogene. 11. 833-840 (1995)
• [Publications] Mitani K: "Generation of the AML1/Evi-1 fusion gene in the t(3;21) (826;822) causes blastic crisis in chronic myelocytic leukemia." EMBO J.13. 504-510 (1994)
• [Publications] Tanaka T: "Evi-1 raises AP-1 activity and stimulatesc-fos promoter transactivation with dependence on the second zinc finger domain." J.Biol.Chem. 269. 24020-24026 (1994)
• [Publications] Ogawa S: "Homozygous loss of the cyclin-dependent kinase 4-inhibitor(p16) gene in human leukemias." Blood. 84. 2431-2435 (1994)
• [Publications] Tanaka T: "An acute myeloid leukemia gene,AML1,regulates hemopoietic myeloid cell differentiation and transcriptional activation antagohistically by two alternative spliced forms." EMBO J. 14. 341-350 (1994)
• [Publications] Tanaka T: "Dual functions of the AML1/Evi-1 chimericprotein in the mechanism of leukemogenesis in t(3;21) leukemias." Mol.Cell.Biol. (in press).
• [Publications] Mitani K: "Cloning of several spieces of MLL/MEN chimeric cDNAs in myeloid leukemias with t(11;19) (823;913.1) translocation." Blood. (in press).
• [Publications] 平井 久丸: "白血病の分子医学" 羊土社, 145 (1994)
• [Publications] 平井 久丸: "臨床分子生物学" 日本臨床社, 874 (1994)