1999 年度実績報告書

造血細胞の遺伝子操作

研究課題

研究課題/領域番号	10181105
研究機関	自治医科大学
研究代表者	小澤敬也自治医科大学, 医学部, 教授 (30137707)
研究分担者	生田宏一京都大学, 医学研究科, 助教授 (90193177) 吉田進昭東京大学, 医科学研究所, 教授 (10250341) 谷憲三朗東京大学, 医科学研究所, 助教授 (00183864) 竹田潤二大阪大学, 医学部, 教授 (50163407)
キーワード	遺伝子治療 / 造血幹細胞 / 選択的増幅遺伝子 / コモンマーモセット / コンディショナルターゲティング / サイトカイン / V-J組換え / 発作性夜間血色素尿症(PNH)
研究概要	体細胞あるいは生殖系列細胞の遺伝子操作を軸とした造血システムの研究を行った。 [造血幹細胞の直接的遺伝子操作] 1. 遺伝子導入造血幹細胞の選択的増幅システムの開発(小澤) : これまで開発を進めてきた選択的増幅遺伝子及びその改良型の評価、さらにMpl遺伝子を利用した新規選択的増幅遺伝子の開発を行った。 2. 小型霊長類造血幹細胞の特性解明と遺伝子導入(谷) : マーモセット移植抗原クラスII遺伝子多型性に基づきコモンマーモセットの家系図を作製すると共に、クラスI遺伝子がマーモセット染色体第4長腕セントロメア近傍にあることを明らかにした。 [発生工学的アプローチによる造血システムの解析] 1. 発生工学的手法を用いた造血系システムの解析(吉田) : コンディショナルターゲティングによりT細胞特異的STAT3欠損マウスを作成し、正常T細胞においてIL-6やIL-2刺激による増殖反応が著明に低下していること、その機構は異なっていることを明らかにした。 2. 造血幹細胞へのIL-7受容体(IL-7R)遺伝子の導入(生田) : IL-7Rにより活性化されたStat5がJγ遺伝子のgermline転写を誘導し、転写共役因子によるヒストンのアセチル化を介してTCRγ遺伝子座のaccessibilityを上昇させ、V-J組換えを誘導することを明らかにした。 3. 発作性夜間血色素尿症(PNH)クローンの増殖優位性獲得機序の解明(竹田) : コンディショナルジーンターゲティング法により血液系のみで遺伝子破壊がみられるマウスシステムを構築し、発作性夜間血色素尿症(PNH)の病態を解析した。PIG-A変異だけではPNHの病態を説明できないことが判明した。

研究成果
(30件)

すべてその他

すべて文献書誌 (30件)

[文献書誌] Matsuda, K. M.: "A novel strategy for the tumor anglogenesis-targeted gene therapy : Generation of angiostatin endogenous plasminogen plasminogen by protease gene transfer"Cancer Gene Ther.. (in press).
[文献書誌] Urabe, M.: "DNA/calcium phosphase precipirates mixed with medium are stable and maintain high transduction efficiency"Anal. Biochem.. 278. 91-92 (2000)
[文献書誌] Maeda, Y.: "Endogenously generated nitric oxide by nitric-oxide synthase gene transfer inhibits cellular proliferation"J. Phamacol. Exp. Therapeut.. 292. 387-393 (2000)
[文献書誌] Ogasawara, Y.: "Potential application of domain negative retinoic acid receptor genes for ex vivo expansion of hematopoietic stem cells"Gene Ther. Mol. Biol.. 3. 293-300 (1999)
[文献書誌] Ogasawara, Y.: "Highly-regulated expression of adeno-associated virus large Rep proteins in stable 293 cell lines using the Cre-loxP switching system"J. Gen. Virol.. 80. 2477-2480 (1999)
[文献書誌] Xu, R.: "A selective amplifier gene for tamoxifen-inducible expansion of hematopoietic cells"J. Gene Med.. 1. 236-244 (1999)
[文献書誌] Matsuda, K. M.: "Development of a modified selective a mplifier gene for hematopoietic stem cell gene therapy"Gene Ther.. 6. 1038-1044 (1999)
[文献書誌] Kume, A.: "Green fluorescent protein as a selectable marker of retrovirally tranduced hematopoietic progenitors"Stem Cells. 17. 226-232 (1999)
[文献書誌] Kume, A.: "A G-CSF receptor-gyrase B fusion gene : a new type of molecular switch for expansion of genetically modified hematopoietic cells"Biochem. Biophs. Res. Commun.. 260. 9-12 (1999)
[文献書誌] Ozawa, K.: "Strategies for gene therapy of Parkinson's disease using adeno-associated virus(AAV) vectors"Biogenic Amines15. 15. 21-37 (1999)
[文献書誌] Ogasawara, Y.: "Effficient production of adeno-associated virus vectors using spilit-type helper plasmids"Jpn. J. Cancer Res.. 90. 476-483 (1999)
[文献書誌] Urabe, M.: "Changed-to-alanine scanning mutagenesis of the N-teminal half of adeno-associated virus type 2 Rep 78 Protein"J. Virol.. 73. 2682-2693 (1999)
[文献書誌] Watari, K.: "Identification of a melanoma antigen, PRAME, as a BCR/ABL-inducible gene"FEBS lett.. 466. 367-371 (2000)
[文献書誌] Hibino, H.: "Common Marmoset as a Target Preclinical Primate for Cytokine and Gene Therapy Studies"Blood. 93. 2839-2848 (1999)
[文献書誌] Koseki, S.: "Factors governing the activity in vino of ribozymes transcribed by RNA polymerase III"J. Virol. 73. 1868-1877 (1999)
[文献書誌] Turuta, T.: "Alkaline phosphatase, defensin gene expression and effect of myeloid cell growth factors in normal and leukemic cells"Leuk. Lymphoma. 32. 237-247 (1999)
[文献書誌] Tsuruta, T.: "Myeloperoxidase gene expression and regulation by myeloid cell growth factors in normal and leukemic cells"Leuk. Lymphoma. 32. 257-267 (1999)
[文献書誌] Kawai, K.: "Gene-modified immunotherapy for renal cell carcinoma"Contrib. Nephrol.. 128. 75-81 (1999)
[文献書誌] Kaisho, T.: "Impairment of natural killer cytotoxic activity and interferon-γ production in C/EBPγ-deficient mice"J. Exp. Med.. 190. 1573-1582 (1999)
[文献書誌] Hizaki, H.: "Abortive expansion of the cumulus and impaired fertility in mice lacking the the prostaglandin E receptor subtype EP2"Proc. Natl. Acad. Sci. , USA. 96. 10501-10506 (1999)
[文献書誌] Eguchi, N.: "Lack of tactile pain(allodynia) in lipocalin-type prostaglandin D synthese-deficient mice"Proc. Natl. Acad. Sci. , USA. 96. 726-730 (1999)
[文献書誌] Kodaira, Y.: "Antigen-driven clonal accumulation of peritoneal γδ T cells in vivo"Immunological Investigations. 28. 137-148 (1999)
[文献書誌] Shinkura, R.: "Alymphoplasia(aly) is caused by a point mutation in the mouse NF-kB inducing kinase gene"Nat. Genet.. 22. 74-77 (1999)
[文献書誌] Yoshida, H.: "IL-7 receptor a^+ CD^<3-> cells in the embryonic intestine induces the organization center or Peyer's patches"Int. Immunol.. 11. 643-655 (1999)
[文献書誌] Sakiyama, T.: "Requirement of IL-5 for induction of autoimmune hemolytic anemia in anti-red blood cell autoantibody trangenic mice"Int. Immunol. 11. 995-1000 (1999)
[文献書誌] Ye, S.-K.: "Induction of germline transcription in the TCRγ locus by Stat5 : implications for accessibility control by the IL-7 receptor"Immunity. 11. 213-223 (1999)
[文献書誌] Nozaki, M.: "Developmental abnormalities of glycosylphosphatidylinositol-anchor deficient embryoys revealed by Cre/loxP system"Lab. Invest.. 79. 293-299 (1999)
[文献書誌] Murakami, Y.: "Different roles of glycosylphosphatidylinositol in various hematopoietic cells revealed by model mice of paroxysmal noctumal hemoglobinuria"Blood. 94. 2963-2970 (1999)
[文献書誌] Inoue, N.: "Human and mouse GPAAI(Glycosylphosphatidylinositol anchoe attachment 1) genes : genomic structures, chromosome loci and the presence of a minor class intron"Cytogenet. Cell. Genrt.. 84. 199-205 (1999)
[文献書誌] Kondo, G.: "Tissue-inherent fate of GPI revealed by GPI-anchored GFP transgenesis"FEBS lett.. 458. 299-303 (1999)

1999 年度 実績報告書

造血細胞の遺伝子操作

研究代表者

小澤 敬也 自治医科大学, 医学部, 教授 (30137707)

研究成果

[文献書誌] Matsuda, K. M.: "A novel strategy for the tumor anglogenesis-targeted gene therapy : Generation of angiostatin endogenous plasminogen plasminogen by protease gene transfer"Cancer Gene Ther.. (in press).

[文献書誌] Urabe, M.: "DNA/calcium phosphase precipirates mixed with medium are stable and maintain high transduction efficiency"Anal. Biochem.. 278. 91-92 (2000)

[文献書誌] Maeda, Y.: "Endogenously generated nitric oxide by nitric-oxide synthase gene transfer inhibits cellular proliferation"J. Phamacol. Exp. Therapeut.. 292. 387-393 (2000)

[文献書誌] Ogasawara, Y.: "Potential application of domain negative retinoic acid receptor genes for ex vivo expansion of hematopoietic stem cells"Gene Ther. Mol. Biol.. 3. 293-300 (1999)

[文献書誌] Ogasawara, Y.: "Highly-regulated expression of adeno-associated virus large Rep proteins in stable 293 cell lines using the Cre-loxP switching system"J. Gen. Virol.. 80. 2477-2480 (1999)

[文献書誌] Xu, R.: "A selective amplifier gene for tamoxifen-inducible expansion of hematopoietic cells"J. Gene Med.. 1. 236-244 (1999)

[文献書誌] Matsuda, K. M.: "Development of a modified selective a mplifier gene for hematopoietic stem cell gene therapy"Gene Ther.. 6. 1038-1044 (1999)

[文献書誌] Kume, A.: "Green fluorescent protein as a selectable marker of retrovirally tranduced hematopoietic progenitors"Stem Cells. 17. 226-232 (1999)

[文献書誌] Kume, A.: "A G-CSF receptor-gyrase B fusion gene : a new type of molecular switch for expansion of genetically modified hematopoietic cells"Biochem. Biophs. Res. Commun.. 260. 9-12 (1999)

[文献書誌] Ozawa, K.: "Strategies for gene therapy of Parkinson's disease using adeno-associated virus(AAV) vectors"Biogenic Amines15. 15. 21-37 (1999)

[文献書誌] Ogasawara, Y.: "Effficient production of adeno-associated virus vectors using spilit-type helper plasmids"Jpn. J. Cancer Res.. 90. 476-483 (1999)

[文献書誌] Urabe, M.: "Changed-to-alanine scanning mutagenesis of the N-teminal half of adeno-associated virus type 2 Rep 78 Protein"J. Virol.. 73. 2682-2693 (1999)

[文献書誌] Watari, K.: "Identification of a melanoma antigen, PRAME, as a BCR/ABL-inducible gene"FEBS lett.. 466. 367-371 (2000)

[文献書誌] Hibino, H.: "Common Marmoset as a Target Preclinical Primate for Cytokine and Gene Therapy Studies"Blood. 93. 2839-2848 (1999)

[文献書誌] Koseki, S.: "Factors governing the activity in vino of ribozymes transcribed by RNA polymerase III"J. Virol. 73. 1868-1877 (1999)

[文献書誌] Turuta, T.: "Alkaline phosphatase, defensin gene expression and effect of myeloid cell growth factors in normal and leukemic cells"Leuk. Lymphoma. 32. 237-247 (1999)

[文献書誌] Tsuruta, T.: "Myeloperoxidase gene expression and regulation by myeloid cell growth factors in normal and leukemic cells"Leuk. Lymphoma. 32. 257-267 (1999)

[文献書誌] Kawai, K.: "Gene-modified immunotherapy for renal cell carcinoma"Contrib. Nephrol.. 128. 75-81 (1999)

[文献書誌] Kaisho, T.: "Impairment of natural killer cytotoxic activity and interferon-γ production in C/EBPγ-deficient mice"J. Exp. Med.. 190. 1573-1582 (1999)

[文献書誌] Hizaki, H.: "Abortive expansion of the cumulus and impaired fertility in mice lacking the the prostaglandin E receptor subtype EP2"Proc. Natl. Acad. Sci. , USA. 96. 10501-10506 (1999)

[文献書誌] Eguchi, N.: "Lack of tactile pain(allodynia) in lipocalin-type prostaglandin D synthese-deficient mice"Proc. Natl. Acad. Sci. , USA. 96. 726-730 (1999)

[文献書誌] Kodaira, Y.: "Antigen-driven clonal accumulation of peritoneal γδ T cells in vivo"Immunological Investigations. 28. 137-148 (1999)

[文献書誌] Shinkura, R.: "Alymphoplasia(aly) is caused by a point mutation in the mouse NF-kB inducing kinase gene"Nat. Genet.. 22. 74-77 (1999)

[文献書誌] Yoshida, H.: "IL-7 receptor a^+ CD^<3-> cells in the embryonic intestine induces the organization center or Peyer's patches"Int. Immunol.. 11. 643-655 (1999)

[文献書誌] Sakiyama, T.: "Requirement of IL-5 for induction of autoimmune hemolytic anemia in anti-red blood cell autoantibody trangenic mice"Int. Immunol. 11. 995-1000 (1999)

[文献書誌] Ye, S.-K.: "Induction of germline transcription in the TCRγ locus by Stat5 : implications for accessibility control by the IL-7 receptor"Immunity. 11. 213-223 (1999)

[文献書誌] Nozaki, M.: "Developmental abnormalities of glycosylphosphatidylinositol-anchor deficient embryoys revealed by Cre/loxP system"Lab. Invest.. 79. 293-299 (1999)

[文献書誌] Murakami, Y.: "Different roles of glycosylphosphatidylinositol in various hematopoietic cells revealed by model mice of paroxysmal noctumal hemoglobinuria"Blood. 94. 2963-2970 (1999)

[文献書誌] Inoue, N.: "Human and mouse GPAAI(Glycosylphosphatidylinositol anchoe attachment 1) genes : genomic structures, chromosome loci and the presence of a minor class intron"Cytogenet. Cell. Genrt.. 84. 199-205 (1999)

[文献書誌] Kondo, G.: "Tissue-inherent fate of GPI revealed by GPI-anchored GFP transgenesis"FEBS lett.. 458. 299-303 (1999)

1999 年度実績報告書

小澤敬也自治医科大学, 医学部, 教授 (30137707)