HIVベクターの開発

• Project/Area Number: 12217142
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Nippon Medical School
• Principal Investigator: 島田 隆 日本医科大学, 大学院・医学研究科, 教授 (20125074)
• Co-Investigator(Kenkyū-buntansha): 塙 秀樹 日本医科大学, 医学部, 助手 (10256977) ; 三宅 弘一 日本医科大学, 医学部, 講師 (90267211)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥55,300,000 (Direct Cost: ¥55,300,000); Fiscal Year 2004: ¥11,000,000 (Direct Cost: ¥11,000,000); Fiscal Year 2003: ¥11,100,000 (Direct Cost: ¥11,100,000); Fiscal Year 2002: ¥11,000,000 (Direct Cost: ¥11,000,000); Fiscal Year 2001: ¥11,200,000 (Direct Cost: ¥11,200,000); Fiscal Year 2000: ¥11,000,000 (Direct Cost: ¥11,000,000)
• Keywords: 遺伝子治療 / ウイルスベクター / HIVベクター / 挿入変異 / 血管新生 / リンパ腫 / 多発性骨髄腫 / 膵癌 / 血管新生阻害 / エンドスタチン / アンギオスタチン / 白血病 / 非分裂細胞 / 血管新生抑制因子 / 骨髄幹細胞 / アデノウイルスベクター / エイズ / ATL

Research Abstract

HIVベクターの開発と癌の遺伝子治療の可能性について検討した。
1)HIVベクターの改良:レトロウイルスベクターによる白血病の報告を受け、HIVベクターの挿入変異による細胞の癌化の可能性を低下させるための研究を進めている。HIVベクターの内部プロモーターが周囲の遺伝子を活性化する度合いを測定するアッセイ系を確立し、内部プロモーターとインスレーターの影響を検討した。内部プロモーターとしては、イントロンをもつEF1aのプロモータを逆向きに挿入したベクターが最も周囲への影響が少なかった。また、chicken globin遺伝子のコア・インスレーター(cHS4)を発現ユニットの両側に挿入することで、ベクターのタイターを下げることなく周囲への影響を遮断できることを見出した。
2)癌遺伝子治療法の開発:血管新生抑制作用のあるendostatinをもつウイルスベクターHIV-EndあるいはAAV-Endを作製し癌遺伝子治療の可能性を検討した。EB陽性ヒトリンパ腫細胞の増殖が筋肉内に投与したHIV-Endにより著明に抑制され、生存曲線の改善も認められた。同様にSCIDマウスに移植された多発性骨髄腫細胞の増殖が造血幹細胞に導入したHIV-Endにより著明に抑制されることが明らかした。同所移植の系として、膵癌細胞をハムスターの膵臓に移植し肝臓転移を起こすモデルを作製し、AAV-Endが原発腫瘍の増殖、及び転移の両者に著明な抑制効果をもつことを示した。また、我々が新たに作成した慢性白血病を自然発症するbcr/ablトランスジェニックマウスをモデルとしたendostatinの遺伝子治療の有効性を確認した。

Research Products

• [Journal Article] Antiangiogenic gene therapy of myeloproliferative disease developed in transgenic mice expressing P230 ber/abl. (2005)
  • Author(s): Miyake, K., Inokuchi, K., et al.
  • Journal Title: Gene Ther. (in press)
• [Journal Article] Near-triploidy and niar-tetraploidy in hematological malignancies and mutation of the p53 gene. (2004)
  • Author(s): Watanabe, A., Inokuchi, K., et al.
  • Journal Title: Clin.Lab.Haem. 26 Pages: 25-30
• [Journal Article] HIV vector mediated intra-articular expression of angiostatin inhibits progression of collagen-induced arthritis in mice. (2004)
  • Author(s): Kato, K., Miyake, K., et al.
  • Journal Title: Rheumatol.Int. (in press)
• [Journal Article] Adeno-associated viral vector mediated expression on endostatin inhibits tumor growth and metastasis in an orthotropic pancreatic cancer model in hamsters. (2004)
  • Author(s): Noro, T., Miyake, K., et al.
  • Journal Title: Cancer Res. 64 Pages: 7486-7490
• [Journal Article] Lentivirus-mediated expression of angiostatin efficiently inhibits neovascularization in a murine proliferative retinopathy model. (2003)
  • Author(s): Igarashi, T., Miyake, K., et al.
  • Journal Title: Gene Ther. 10 Pages: 219-226
• [Journal Article] Myeloproliferative diseasein transgenic mice expressing P230 Bcr/Able : longer disease latency, thrombocytosis and mild leukocytosis. (2003)
  • Author(s): Inokuchi, K., Dan, K., et al.
  • Journal Title: Blood 102 Pages: 320-322
• [Journal Article] Simultaneous novel BCR-ABL gene mutation and increased expression of BCR-ABL mRNA caused clinical resistence in a double Ph-positive acute biphenotypic leukemia. (2003)
  • Author(s): Inokuchi, K., Inami, M., et al.
  • Journal Title: Int.J.Hematol. 78 Pages: 173-175
• [Publications] Okino, T., et al.: "Sequential hitopathological changes in vivo after suicide gene therapy of gastric cancer induced by N-methyl-N'-nitro-N-nitroguanidine in rats."Jpn.J.Cancer Res.. 92. 673-679 (2001)
• [Publications] Inokuchi, K., et al.: "Loss of DCC gene expression is of prognostic importance in acute myelogenous leukemia."Clinical Cancer Res.. 8. 1882-1888 (2002)
• [Publications] Igarashi, T., et al.: "New strategy for in vivo transgene expression in corneal epithelial progenitor cells."Current Eye Res.. 24. 46-50 (2002)
• [Publications] Igarashi, T., et al.: "Lentivirus-mediated expression of angiostatin efficiently inhibits neovascularization in a murine proliferative retinopathy model."Gene Ther.. 10. 219-226 (2003)
• [Publications] Inokuchi, K., et al.: "Myeloproliferative disease in transgenic mice expressing P230 Bcr/Able : longer disease latency, thrombocytosis and mild leukocytosis."Blood. 102. 320-322 (2003)
• [Publications] Watanabe, A., et al.: "Near-triploidy and niar-tetraploidy in hematological malignancies and mutation of the p53 gene."Clin.Lab.Haem.. 26. 25-30 (2004)
• [Publications] Inokuchi, K., Yamaguchi, H., Hanawa, H., Tanosaki, et al.: "Loss of DCC gene expression is of prognostic importance in acute myelogenous leukemia"Clinical Cancer Res.. 8. 1882-1888 (2002)
• [Publications] Igarashi, T., Miyake, K., Kato, K., Watanabe, A., et al.: "Lentivirus-mediated expression of angiostatin efficiently inhibits neovascularization in a murine proliferative retinopathy model"Gene Ther.. 10. 219-226 (2003)
• [Publications] Igarashi, T., Miyake, K., Suzuki, N., Kato, K., et al.: "New strategy for in vivo transgene expression in corneal epithelial progenitor cells"Current Eye Res.. 24. 46-50 (2002)
• [Publications] Takahashi, H., Hirai, Y., Migita, M., Seino, Y., et al.: "Long-term systemic therapy of Fabry disease in a knockout mouse by adeno-associated virus-mediated muscle-directed gene transfer"Proc. Natl. Acad. Sci. USA.. 99. 13777-13782 (2002)
• [Publications] Tanaka, R., Komine-Kobayashi, M., Mochizuki, H., Yamada, et al.: "Migration of EGFP expressing bone marrow-derived microglia/macrophage into the mice brain following permanent focal ischemia"Neuroscience. (in press). (2003)
• [Publications] Sugiyama, O., Orimo, H., Suzuki, S., Yamashita, K., et al.: "Bone formation following tansplantation of genetically modified primary bone marrow stromal cells"J. Orthop. Res.. (in press). (2003)
• [Publications] Miyake, K., Shimada, T., et al.: "Selective killing of HIV infected cells by targeted gene transfer and inducible gene expression using a recombinant HIV vector"Hum. Gene. Ther. 12. 227-233 (2001)
• [Publications] Sakai, N., Shimada, T., et al.: "Selective transduction of HIV-1 infected cells by the combination of HIV and MMLV vectors"Int. J. Hematol.. 73. 476-482 (2001)
• [Publications] Mochizuki, H., Shimada, T., et al.: "An AAV-derived Apaf-1 dominant negative inhibitor prevents MPTP toxicity as antiapoptotic gene therapy for Parkinson's disease"Proc. Natl. Acad. Sci. U.S.A. 98. 10918-10923 (2001)
• [Publications] Nakano, K., Shimada, T., et al.: "Differentiation of transplanted bone marrow cells in the adult mouse brain"Transplantation. 71. 1735-1740 (2001)
• [Publications] Okino, T., Shimada, T., et al.: "Sequential histopathological changes in vivo after suicide gene therapy of gastric cancer induced by N-nitro in rats"Jpn. J. Cancer Res.. 92. 673-679 (2001)
• [Publications] Akasaka, S., Shimada, T., et al.: "Suicide gene therapy for chemically induced rat bladder tumor entailing instillation of adenoviral vectors"Jpn. J. Cancer Res.. 92. 568-575 (2001)
• [Publications] Miyake,K.,Shimada,T., et al.: "Selectivekilling of HIV infectedcells by targeted gene transfer and inducible gene expression using a recombinant HIV vector."Hum.Gene Ther.. 112. 227-233 (2001)
• [Publications] Nakano,K.,Shimada,T., et al.: "Differentiation of transplanted bone marrow cells in the adult mouse brain."Transplantation. (in press). (2001)
• [Publications] Sakai,N.,Shimada,T., et al.: "Selective transduction of HIV-1 infected cells by the combination of HIV and MLV vectors."Inter.J.Hematol.. (in press). (2001)
• [Publications] Fujii,I.,Shimada,T., et al.: "Targeted and stable gene delivery into muscle cells by a two-step transfer system."Biochem.Biophys.Res.Commun.. 275. 931-935 (2001)
• [Publications] Satoh,W.,Shimada,T., et al.: "Site-specific integration of an AAV vector plasmid mediated by regulated expression of Rep based on Cre/lox-p recombination."J.Virol.. 74. 10631-10638 (2000)
• [Publications] Suzuki,S.and Shimada,T.: "A retroviral vector capable of targeted gene transfer into cells expressing HIV envelope glycoprotein."Biochem.Biophys.Res.Commun.. 271. 672-676 (2000)