1993 年度実績報告書

がんに対する細胞移植/遺伝子治療の基礎的研究

研究課題

研究課題/領域番号	05151019
研究機関	東京大学
研究代表者	浅野茂隆東京大学, 医科学研究所, 教授 (50134614)
研究分担者	小澤敬也東京大学, 医科学研究所, 助教授 (30137707) 渡部好彦京都大学, 薬学部, 助教授 (90109075) 新津洋司郎札幌医科大学, 教授 (10045502) 斉藤英彦名古屋大学, 医学部, 教授 (20153819) 内山卓京都大学, ウイルス研, 教授 (80151900)
キーワード	がん / 遺伝子治療 / 遺伝子導入 / レトロウイルスベクター / アデノウイルスベクター / リポソーム法 / 細胞移植 / 抗腫瘍免疫
研究概要	1.遺伝子導入法の基礎検討:レトロウイルスベクターとアデノウイルスベクターの比較検討を行ったところ、後者は遺伝子発現持続期間が短いものの、遺伝子導入効率は優れていた。また、HVJ-リポソーム法によりアンチセンスオリゴヌクレオチドを血管平滑筋細胞へ効率良く導入する方法を確立し、in vivo遺伝子導入への応用が可能であることを示した。 2.癌細胞への遺伝子導入による増殖阻害:白血病細胞にHSV-TK遺伝子を導入し、ガンシクロビル処理による殺細胞効果を確認した。また、転座型白血病ではキメラ遺伝子転写産物の融合点に対するアンチセンスオリゴヌクレオチドやリボザイムの利用に関する検討を進めた。その他、RAS抑制変異体が腫瘍抑制活性を示すことを明らかにした。尚、動物モデルの準備として、SCIDマウスを用いてATL細胞のin vivo増殖系を確立した。 3.癌に対する免疫遺伝子治療の基礎検討: TNF,IFN-γ,GM-CSFなどの遺伝子を導入した腫瘍細胞をマウスに移植し、抗腫瘍免疫能の誘導を図り、免疫学的解析を行った。CTLの誘導にはCD28を介したシグナルが重要であるが、新たなリガンド(B70)のcDNAクローニングに成功した。また、抗原提示細胞として重要な役割を果たす樹状突起細胞の分離培養法(GM-CSFを使用)を確立した。 4.細胞移植療法:骨髄移植後の白血病再発に対して、ドナーリンパ球輸注療法を実施し、一部の症例で有効性が確認された。以上の他に、造血幹細胞や白血病細胞の増殖機構を解明するため、、受容体-シグナル伝達系の解析を進めた。また、相同組換えを利用した遺伝子ターゲティング法に関する研究を行った。

研究成果
(22件)

すべてその他

すべて文献書誌 (22件)

[文献書誌] Nishikawa,M.: "Changes in hematopoiesis-supporting ability of C3H10T1/2 mouse embryo fibroblasts during differentiation." Blood. 81. 1184-1192 (1993)
[文献書誌] Dranoff,G.: "Vaccination with irradiated tumor cells engineered to secrete murine granulocyte-macrophage colony-stimulating factor stimulates potent,specific and llong-lasting anti-tumor immunity." Proc.Natl.Acad.Sci.USA. 90. 3539-3543 (1993)
[文献書誌] Uchiyama,A.: "Transfection of interleukin-2 gene into human melanoma cells augments cellular immune response." Cancer Res.53. 949-952 (1993)
[文献書誌] Morishita,R.: "Intraluminal molecular delivery of antisense CDC2 kinase and PCNA oligonucleotides prevent restenosis after angioplasty." Proc.Natl.Acad.Sci.USA. 90. 8474-8478 (1993)
[文献書誌] Abe,A.: "Transduction of a drug-sensitive toxic gene into human leukemia cell lines with a novel retroviral vector." Toxin Gene for Killing leukemia Cells. 203. 354-359 (1993)
[文献書誌] Masegi,T.: "Lysosome labilizers potentiate the antitumor effects of tumor necrosis factor-α." Jpn.J.Cancer Res.84. 451-454 (1993)
[文献書誌] Azuma,M.: "B70 antigen is a second ligand for CTLA-4 and CD28." Nature. 366. 76-79 (1993)
[文献書誌] Yagita,H.: "Bispecific antibody as a potentiator of tumor cell killing by IL-2-activated lymphocytes." gann Monograph on Cancer Res.40. 95-106 (1993)
[文献書誌] Teramura,Y.: "Interferon-γ-producing tumor induces host tumor-specific T cell responses." Jpn.J.Cancer Res.84. 689-696 (1993)
[文献書誌] Porgador,A.: "Anti-metastatic vaccination of tumor-bearing mice with two types of γ IFN gene inserted tumor cells." J.Immunol.150. 1458-1470 (1993)
[文献書誌] Kondo,A.: "A model of in vivo cell proliferation of adult T-cell leukemia." Blood. 82. 2501-2509 (1993)
[文献書誌] Ogiso,Y.: "Resistance of NIH3T3 cells to v-fes transformation induced by a dominant negative H-ras mutant." Exp.Cell Res.208. 415-421 (1993)
[文献書誌] Miura,N.: "Stem cell factor and IL-6 preferentially support the proliferation of kit-positive hemopoietic progenitors rather than their differentiation." Exp.Hematol.21. 143-149 (1993)
[文献書誌] Watanabe,S.: "Reconstituted human granulocyte-macrophage colony-stimulating factor(GM-CSF)receptor transduces growth promoting signals in mouse NIH3T3 cells:Comparison with signalling in BA/F3 proB cells." Mol.Cell.Biol.13. 1440-1448 (1993)
[文献書誌] Takahashi,S.: "Recombinant human glycosylated granulocyte colony-stimulating factor(rhG-CSF)-combined regimen for a allogeneic bone marrow transplantation in refractory acute myeloid leukemia." Bone Marrow Transplant. (in press).
[文献書誌] Tani,K.: "Retrovirus-mediated gene transfer of human pyruvate kinase(PK)cDNA into murine hematopoietic cells:implications for gene therapy of human PK deficiency." Blood. (in press).
[文献書誌] Kanegae,Y.: "A simple and efficient purification method of infectious adenovirus virions." Jap.J.Med.Sci.Biol.(in press).
[文献書誌] Fujii,S.: "Augmented systemic immunity in mice implanted with TNF-α gene transduced Meth-A cells." Jap.J.Cancer Res.(in press).
[文献書誌] Uemura,N.: "The Mechanism and New Approach on Drug Resistance of Cancer Cells. 13" Elsevier Science Publishers B.V., 1440-1448 (1993)
[文献書誌] Dranoff,G.: "Clinical Pharmacology Vol 9" Frontiers in Biomedical Research, 22-29 (1993)
[文献書誌] Kaneda,Y.: "Methods in Enzymology Vol.221" Academic Press,Inc.,Orlando,Florida, 317-327 (1993)
[文献書誌] Kaneda,Y.: "A Laboratory Handbook" Academic Press,Inc.,Orlando,Florida(in press),

1993 年度 実績報告書

がんに対する細胞移植/遺伝子治療の基礎的研究

研究代表者

浅野 茂隆 東京大学, 医科学研究所, 教授 (50134614)

研究成果

[文献書誌] Nishikawa,M.: "Changes in hematopoiesis-supporting ability of C3H10T1/2 mouse embryo fibroblasts during differentiation." Blood. 81. 1184-1192 (1993)

[文献書誌] Dranoff,G.: "Vaccination with irradiated tumor cells engineered to secrete murine granulocyte-macrophage colony-stimulating factor stimulates potent,specific and llong-lasting anti-tumor immunity." Proc.Natl.Acad.Sci.USA. 90. 3539-3543 (1993)

[文献書誌] Uchiyama,A.: "Transfection of interleukin-2 gene into human melanoma cells augments cellular immune response." Cancer Res.53. 949-952 (1993)

[文献書誌] Morishita,R.: "Intraluminal molecular delivery of antisense CDC2 kinase and PCNA oligonucleotides prevent restenosis after angioplasty." Proc.Natl.Acad.Sci.USA. 90. 8474-8478 (1993)

[文献書誌] Abe,A.: "Transduction of a drug-sensitive toxic gene into human leukemia cell lines with a novel retroviral vector." Toxin Gene for Killing leukemia Cells. 203. 354-359 (1993)

[文献書誌] Masegi,T.: "Lysosome labilizers potentiate the antitumor effects of tumor necrosis factor-α." Jpn.J.Cancer Res.84. 451-454 (1993)

[文献書誌] Azuma,M.: "B70 antigen is a second ligand for CTLA-4 and CD28." Nature. 366. 76-79 (1993)

[文献書誌] Yagita,H.: "Bispecific antibody as a potentiator of tumor cell killing by IL-2-activated lymphocytes." gann Monograph on Cancer Res.40. 95-106 (1993)

[文献書誌] Teramura,Y.: "Interferon-γ-producing tumor induces host tumor-specific T cell responses." Jpn.J.Cancer Res.84. 689-696 (1993)

[文献書誌] Porgador,A.: "Anti-metastatic vaccination of tumor-bearing mice with two types of γ IFN gene inserted tumor cells." J.Immunol.150. 1458-1470 (1993)

[文献書誌] Kondo,A.: "A model of in vivo cell proliferation of adult T-cell leukemia." Blood. 82. 2501-2509 (1993)

[文献書誌] Ogiso,Y.: "Resistance of NIH3T3 cells to v-fes transformation induced by a dominant negative H-ras mutant." Exp.Cell Res.208. 415-421 (1993)

[文献書誌] Miura,N.: "Stem cell factor and IL-6 preferentially support the proliferation of kit-positive hemopoietic progenitors rather than their differentiation." Exp.Hematol.21. 143-149 (1993)

[文献書誌] Watanabe,S.: "Reconstituted human granulocyte-macrophage colony-stimulating factor(GM-CSF)receptor transduces growth promoting signals in mouse NIH3T3 cells:Comparison with signalling in BA/F3 proB cells." Mol.Cell.Biol.13. 1440-1448 (1993)

[文献書誌] Takahashi,S.: "Recombinant human glycosylated granulocyte colony-stimulating factor(rhG-CSF)-combined regimen for a allogeneic bone marrow transplantation in refractory acute myeloid leukemia." Bone Marrow Transplant. (in press).

[文献書誌] Tani,K.: "Retrovirus-mediated gene transfer of human pyruvate kinase(PK)cDNA into murine hematopoietic cells:implications for gene therapy of human PK deficiency." Blood. (in press).

[文献書誌] Kanegae,Y.: "A simple and efficient purification method of infectious adenovirus virions." Jap.J.Med.Sci.Biol.(in press).

[文献書誌] Fujii,S.: "Augmented systemic immunity in mice implanted with TNF-α gene transduced Meth-A cells." Jap.J.Cancer Res.(in press).

[文献書誌] Uemura,N.: "The Mechanism and New Approach on Drug Resistance of Cancer Cells. 13" Elsevier Science Publishers B.V., 1440-1448 (1993)

[文献書誌] Dranoff,G.: "Clinical Pharmacology Vol 9" Frontiers in Biomedical Research, 22-29 (1993)

[文献書誌] Kaneda,Y.: "Methods in Enzymology Vol.221" Academic Press,Inc.,Orlando,Florida, 317-327 (1993)

[文献書誌] Kaneda,Y.: "A Laboratory Handbook" Academic Press,Inc.,Orlando,Florida(in press),

1993 年度実績報告書

浅野茂隆東京大学, 医科学研究所, 教授 (50134614)