1998 年度実績報告書

造血幹細胞の遺伝子操作

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	竹田潤二阪大, 医学部, 教授 (50163407) 生田宏一京大, 医学研究科, 助教授 (90193177) 吉田進昭東大, 医科研, 教授 (10250341) 谷憲三朗東大, 医科研, 助教授 (00183864)
キーワード	選択的増幅遺伝子 / コモンマーモセット / 造血幹細胞 / ジーンターゲティング / ケモカイン / サイトカイン / 発作性夜間血色素尿症(PNH) / クローン優位性
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):遺伝子導入細胞の体内増幅のための選択的増幅遺伝子の改良と、造血幹細胞の体外増幅のための分化制御遺伝子の有効性に関する検討を行った。 (2) 小型霊長類血幹細胞への遺伝子導入実験(谷):小型霊長類コモンマーモセット末梢血幹細胞に多剤耐性遺伝子(MDRI)を導入後、自家移植を行い、MDRI遺伝子導入細胞による造血再構築を長期にわたり観察した。移植約3年後も1%の末梢血・骨髄前駆細胞がビンクリスチン耐性コロニーとなった。 [発生工学的アプローチによる造血システムの解析] (1) ジーンターゲティング法による造血関連遺伝子の解析(吉田)。IL-18欠損マウスでは、IFNγ産生低下、Thl細胞の分化障害やNK細胞活性化の低下がみられたが、IL-12とIL-18が協調して働くことがダブルノックアウトマウスの解析より明らかになった。ケモカインPBSF/SDF-Iやその受容体CXCR4欠損マウスの解析により、これらは胎児肝および骨髄での造血、消化管の血管形成に重要であることを明らかにした。 (2) 造血細胞分化におけるIL-7受容体の役割(生田):IL-3やIL-7依存性の血液細胞株において、IL-7Rのシグナル伝達分子のSTAT5がTCRγ遺伝子のgermline転写を誘導し、accessibilityを調節する可能性が示された。 (3) 発作性夜間血色素尿症(PNH)モデルマウスと造血幹細胞制御(竹田):PNHのクローン優位性機構を解析するため、その原因遺伝子PIG-Aが血液系だけで欠損しているモデルマウスを構築した。しかし、この系ではクローン優位性は観察されず、別の要因の関与が示唆された。

研究成果

(28件)

すべてその他

すべて文献書誌 (28件)

[文献書誌] Ogasawara,Y.: "Efficient production of adeno-associated virus vectors using split-type helper plasmids" Jpn.J.Cancer Res.(in press).
[文献書誌] Matsuda,K.M.: "Development of a modified selective amplifier gene for hematopoietic stem cell gene therapy" Gene Ther.(in press).
[文献書誌] Urabe,M.: "Charged-to-alanine scanning mutagenesis of N-terminal half of adeno-associated virus type 2 Rep78 protein" J.Virol.(in press).
[文献書誌] Ogasawara,Y.: "Potential application of dominant negative retinoic acid receptor genes for ex vivo expansion of hematopoietic stem cells" Gene Ther.Mol.Biol.(in press).
[文献書誌] Kodaira,H.: "Fas and mutant estrogen receptor chimeric gene: a novel suicide vector for tamoxifen-inducible apoptosis" Jpn.J.Cancer Res.89. 741-747 (1998)
[文献書誌] Kokubun,M.: "Apoptosis-mediated regulation of recombinant human granulocyte colony-stimulating factor production by genetically engineered fibroblasts" Gene Ther.5. 923-929 (1998)
[文献書誌] Ozawa,K.: "A novel selective amplifier gene for hematopoietic stem cell gene therapy" Cancer Res.Ther.Contr.7. 27-31 (1998)
[文献書誌] Ogasawara,Y.: "The use of heterologous promoters for adeno-associated virus(AAV)protein expression in AAV vector production" Microbiol.Immunol.42. 177-185 (1998)
[文献書誌] Hibino,H.: "Common Marmoset as a Target Preclinical Primate for Cytokine and Gene Therapy Studies" Blood. (in press).
[文献書誌] Sumimoto,H.: "Superiority of interleukin-12-transduced murine lung cancer cells GM-CSF or B7-1(CD80) transfectants for therapeutics antitumor immunity in syngeneic immunocompetent mice" Cancer Gene Therapy. 5. 29-37 (1998)
[文献書誌] Nakazaki,Y.: "Vaccine effect of granulocyte-macrophage colony-stimulating factor or CD80 gene-transduced murine leukemia/lymphoma cells and their cooperative enhancement of antitumor immunity" Gene Therapy. 5. 1355-1362 (1998)
[文献書誌] Kuwabara,T.: "Maxizyme, an allosterically controllable novel tRNA Val-ribozyme,with extremely high and specific activity in cells with a Philadelphia chromosome induces apoptosis only in leukemic cells without affecting normal cells" Mol Cell. 2. 617-627 (1998)
[文献書誌] Takeda,K.: "Stat3 activation is responsible for IL-6-dependent T cell proliferation through preventing apoptosis: Generation and characterization of T cell specific Stat3-deficient mice" J.Immunol.161. 4652-4660 (1998)
[文献書誌] Ushikubi,F.: "Impaired febrile response in mice lacking the prostaglandin E receptor subtype EP3" Nature. 395. 281-284 (1998)
[文献書誌] Tachibana,K.: "The chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract" Nature. 393. 591-595 (1998)
[文献書誌] Fukuyama,H.: "Transgenic expression of Fas in T cells blocks lymphoproliferation but not autoimmune disease in MRL-lpr mice" J.Immunol.160. 3805-3811 (1998)
[文献書誌] Takeda,K.: "Defective NK cell activity and Th1 response in IL-18-deficient mice" Immunity. 8. 383-390 (1998)
[文献書誌] Sakiyama,T.: "Requirement of interleukin-5 for induction of autoimmune hemolytic anemia in the anti-red blood cell autoantibody transgenic mice" Int.Immunol.(in press).
[文献書誌] Kodaira,Y.: "Antigen-drived clonal accumulation of peritoneal γδ T cells in vivo" Immunological Investigations. (in press).
[文献書誌] Adachi,S.: "Essential role of IL-7 receptor α in the formation of Peyer's patch anlage" Int.Immunol. 10. 1-6 (1998)
[文献書誌] Ishii,S.: "Impaired anaphylactic responses with intact sensitivity to endotoxin in mice lacking a platelet-activating factor receptor" J.Exp.Med.187. 1779-1788 (1998)
[文献書誌] Nozaki,M.: "Developmental abnormalities of glycosylphosphatidylinositol-anchor deficient embryos revealed Cre/loxP system" Lab.Invest. (in press).
[文献書誌] Tsujimura,A.: "Molecular cloning of a murine homologue of membrane cofactor protein(MCP,CD46): preferential expression in testicular germ cells" Biochem.J.330. 163-168 (1998)
[文献書誌] Watanabe,R.: "The first step of glycosylphosphatidylinositol biosynthesis is mediated by a complex of PIG-A,PIG-H,PIG-C,and GPI1" EMBO J.17. 877-885 (1998)
[文献書誌] Takahama,Y.: "Functional competence of T cells in the abs ence of GPI-anchored proteins caused by T-cell specific disruption of Pig-a gene" Eur.J.Immunol.28. 2159-2166 (1998)
[文献書誌] Tomita,S.,: "A homologue of Saccharomyces cerevisiae Dpmlp is not sufficient for synthesis of dolichol-phosphate-mannose in mammalian cells" J.Biol.Chem.273. 9249-9254 (1998)
[文献書誌] Takeda,K.: "Stat3 activation is responsible for IL-6-dependent T cell proliferation through preventing apoptosis: Generation and characteraization of T cell specific Stat3-deficient mice" J.Immunol.161. 4652-4660 (1998)
[文献書誌] Akaishi,H.: "Defective IL-2-mediated IL-2 receptor α chain expression in Stat3-deficient T lymphocytes" Int.Immunol.10. 1747-1751 (1998)

1998 年度 実績報告書

造血幹細胞の遺伝子操作

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] Ogasawara,Y.: "Efficient production of adeno-associated virus vectors using split-type helper plasmids" Jpn.J.Cancer Res.(in press).

[文献書誌] Matsuda,K.M.: "Development of a modified selective amplifier gene for hematopoietic stem cell gene therapy" Gene Ther.(in press).

[文献書誌] Urabe,M.: "Charged-to-alanine scanning mutagenesis of N-terminal half of adeno-associated virus type 2 Rep78 protein" J.Virol.(in press).

[文献書誌] Ogasawara,Y.: "Potential application of dominant negative retinoic acid receptor genes for ex vivo expansion of hematopoietic stem cells" Gene Ther.Mol.Biol.(in press).

[文献書誌] Kodaira,H.: "Fas and mutant estrogen receptor chimeric gene: a novel suicide vector for tamoxifen-inducible apoptosis" Jpn.J.Cancer Res.89. 741-747 (1998)

[文献書誌] Kokubun,M.: "Apoptosis-mediated regulation of recombinant human granulocyte colony-stimulating factor production by genetically engineered fibroblasts" Gene Ther.5. 923-929 (1998)

[文献書誌] Ozawa,K.: "A novel selective amplifier gene for hematopoietic stem cell gene therapy" Cancer Res.Ther.Contr.7. 27-31 (1998)

[文献書誌] Ogasawara,Y.: "The use of heterologous promoters for adeno-associated virus(AAV)protein expression in AAV vector production" Microbiol.Immunol.42. 177-185 (1998)

[文献書誌] Hibino,H.: "Common Marmoset as a Target Preclinical Primate for Cytokine and Gene Therapy Studies" Blood. (in press).

[文献書誌] Sumimoto,H.: "Superiority of interleukin-12-transduced murine lung cancer cells GM-CSF or B7-1(CD80) transfectants for therapeutics antitumor immunity in syngeneic immunocompetent mice" Cancer Gene Therapy. 5. 29-37 (1998)

[文献書誌] Nakazaki,Y.: "Vaccine effect of granulocyte-macrophage colony-stimulating factor or CD80 gene-transduced murine leukemia/lymphoma cells and their cooperative enhancement of antitumor immunity" Gene Therapy. 5. 1355-1362 (1998)

[文献書誌] Kuwabara,T.: "Maxizyme, an allosterically controllable novel tRNA Val-ribozyme,with extremely high and specific activity in cells with a Philadelphia chromosome induces apoptosis only in leukemic cells without affecting normal cells" Mol Cell. 2. 617-627 (1998)

[文献書誌] Takeda,K.: "Stat3 activation is responsible for IL-6-dependent T cell proliferation through preventing apoptosis: Generation and characterization of T cell specific Stat3-deficient mice" J.Immunol.161. 4652-4660 (1998)

[文献書誌] Ushikubi,F.: "Impaired febrile response in mice lacking the prostaglandin E receptor subtype EP3" Nature. 395. 281-284 (1998)

[文献書誌] Tachibana,K.: "The chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract" Nature. 393. 591-595 (1998)

[文献書誌] Fukuyama,H.: "Transgenic expression of Fas in T cells blocks lymphoproliferation but not autoimmune disease in MRL-lpr mice" J.Immunol.160. 3805-3811 (1998)

[文献書誌] Takeda,K.: "Defective NK cell activity and Th1 response in IL-18-deficient mice" Immunity. 8. 383-390 (1998)

[文献書誌] Sakiyama,T.: "Requirement of interleukin-5 for induction of autoimmune hemolytic anemia in the anti-red blood cell autoantibody transgenic mice" Int.Immunol.(in press).

[文献書誌] Kodaira,Y.: "Antigen-drived clonal accumulation of peritoneal γδ T cells in vivo" Immunological Investigations. (in press).

[文献書誌] Adachi,S.: "Essential role of IL-7 receptor α in the formation of Peyer's patch anlage" Int.Immunol. 10. 1-6 (1998)

[文献書誌] Ishii,S.: "Impaired anaphylactic responses with intact sensitivity to endotoxin in mice lacking a platelet-activating factor receptor" J.Exp.Med.187. 1779-1788 (1998)

[文献書誌] Nozaki,M.: "Developmental abnormalities of glycosylphosphatidylinositol-anchor deficient embryos revealed Cre/loxP system" Lab.Invest. (in press).

[文献書誌] Tsujimura,A.: "Molecular cloning of a murine homologue of membrane cofactor protein(MCP,CD46): preferential expression in testicular germ cells" Biochem.J.330. 163-168 (1998)

[文献書誌] Watanabe,R.: "The first step of glycosylphosphatidylinositol biosynthesis is mediated by a complex of PIG-A,PIG-H,PIG-C,and GPI1" EMBO J.17. 877-885 (1998)

[文献書誌] Takahama,Y.: "Functional competence of T cells in the abs ence of GPI-anchored proteins caused by T-cell specific disruption of Pig-a gene" Eur.J.Immunol.28. 2159-2166 (1998)

[文献書誌] Tomita,S.,: "A homologue of Saccharomyces cerevisiae Dpmlp is not sufficient for synthesis of dolichol-phosphate-mannose in mammalian cells" J.Biol.Chem.273. 9249-9254 (1998)

[文献書誌] Takeda,K.: "Stat3 activation is responsible for IL-6-dependent T cell proliferation through preventing apoptosis: Generation and characteraization of T cell specific Stat3-deficient mice" J.Immunol.161. 4652-4660 (1998)

[文献書誌] Akaishi,H.: "Defective IL-2-mediated IL-2 receptor α chain expression in Stat3-deficient T lymphocytes" Int.Immunol.10. 1747-1751 (1998)

1998 年度実績報告書

小澤敬也自治医科大学, 医学部, 教授 (30137707)