腫瘍の特異的標的化を目指した遺伝子治療法の開発

• Project/Area Number: 06283227
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Research Institution: Sapporo Medical University (1999); Japanese Foundation For Cancer Research (1994-1998)
• Principal Investigator: 濱田 洋文 札幌医科大学, 医学部, 教授 (00189614)
• Co-Investigator(Kenkyū-buntansha): 角田 力弥 福島県立医大, 解剖学2, 助教授 (40045779) ; 山口 聡 (山口 聰) 財団法人癌研究會, 癌化学療法センター・分子生物治療研究部, 研究員 (00280628) ; 花田 賢一 財団法人癌研究會, 癌化学療法センター・分子生物治療研究部, 研究員 (70280625) ; 中村 佳子 財団法人癌研究會, 癌化学療法センター・分子生物治療研究部, 研究員
• Project Period (FY): 1999
• Project Status: Completed (Fiscal Year 1999)
• Budget Amount: ¥88,000,000 (Direct Cost: ¥88,000,000); Fiscal Year 1999: ¥15,000,000 (Direct Cost: ¥15,000,000); Fiscal Year 1998: ¥15,000,000 (Direct Cost: ¥15,000,000); Fiscal Year 1997: ¥15,000,000 (Direct Cost: ¥15,000,000); Fiscal Year 1996: ¥15,000,000 (Direct Cost: ¥15,000,000); Fiscal Year 1995: ¥14,000,000 (Direct Cost: ¥14,000,000); Fiscal Year 1994: ¥14,000,000 (Direct Cost: ¥14,000,000)
• Keywords: adenoviral vector / cancer gene therapy / fiber mutant / integrin / melanoma / melanocyte stimutating hormone / malignant melanoma / gastrin releasing peptide / 遺伝子治療 / アデノウイルスベクター / 腫瘍抑制遺伝子 / 悪性神経膠腫 / 脳腫瘍 / 悪性黒色腫 / EIA / p53 / 免疫制御 / T細胞活性化 / アポトーシス / ファイバー / グリオーマ / 免疫遺伝子治療 / 自殺遺伝子導入療法 / レトロウイルス / アデノウイルス / 抗原提示細胞 / 癌免疫 / サイトカイン遺伝子 / サイトカイン / 腫瘍ワクチン / GM-CSF / IL-4 / レトロウイルス・ベクター / CD82 / ウイルスベクター / 腫瘍ワクチン療法 / 養子免疫療法 / 樹状細胞

Research Abstract

当研究では、細胞側の受容体分子との吸着を担うファイバータンパクに変異を導入することによって、癌細胞の標的化が可能な変異アデノウイルスベクター、Adv-F/K20、Adv-F/MSH、Adv-F/GRP、Adv-F/RGD、Adv-F40Sなどを作成した。
ファイバーに20個のリジンを付加したF/K20変異型は、ヒト脳腫瘍の対して従来型に比べて数十倍高い遺伝子導入効率が得られ、動物実験でも著明な治療効果増強を示した。これをp53プラスp33ING1によるアポトーシス誘導治療に応用したり、腫瘍特異的な制限増幅型のベクターと組み合わせて、グリオーマに高い治療効果が得られることを報告し、脳外科のグループと協力して、F/K20を用いた臨床研究を準備している。
悪性黒色腫、口腔癌などでは、アデノウイルスの受容体CARの発現がきわめて低く、従来型のアデノウイルスベクターでは遺伝子導入効率が非常に低い。これに対し、メラノサイト刺激ホルモン(MSH)を融合させたF/MSH変異ウイルスは、ヒト悪性黒色腫に従来型よりも十倍程度高い遺伝子導入効率を示した。ガストリン放出ペプチド(GRP)を融合させたF/GRP変異ウイルスは、GRP受容体を高発現するヒト前立腺癌などに高い遺伝子導入効率を示した。一方、インテグリンを標的としたRGDモチーフをファイバーのHIループに含むF/RGDウイルスを用いると、悪性黒色腫、口腔の扁平上皮癌などに従来型に比べて数十倍高い遺伝子導入効率が得られることがわかり、臨床への応用が有望である。
また、通常用いられるヒト5型アデノウイルス(Ad5)のファイバーのみを40型の短ファイバーに置き換えたAdv-F40Sベクターは、Ad5のファイバーの受容体CARと結合しない。現在、Adv-F40Sベクターをベースとして各種のリガンドとの組み合わせによって、宿主細胞域の狭まった特異性の高い「がん標的化ウイルスベクター」の作成を目指している。

Research Products

• [Publications] Shinoura, N., et al. And Hamada. H: "Relative level of expression of Bax and Bcl-XL determines the cellular fate of apoptosis/necrosis induced by the overexpression of Bax"Oncogene. 18. 5703-5713 (1999)
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• [Publications] Shinoura, N., et al. And Hamada. H: "Expression level of Bcl-2 determines anti-or pro-apoptotic function"Cancer Res. 59. 4119-4128 (1999)
• [Publications] Shibagaki, N., et al. And Hamada. H: "Overexpression of CD82 on human T cells enhances LFA1/ICAM1-mediated cell-cell adhesion : functional association between CD82 and LFA1 in T cell activation"Eur J Immunol. 29. 4081-4091 (1999)
• [Publications] Motoi, F., et al. And Hamada. H: "Effective gene therapy for pancreatic cancer by cytokines mediated by restricted replication-compelent adenovirus"Hum. Gene Ther. 11(in press). (2000)
• [Publications] Shinoura, N., et al. And Hamada. H: "Adenovirus-mediated transfer of p33ING with p53 drastically augments apoptosis in gliomas"Cancer Res. 59. 5521-5528 (1999)
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• [Publications] Shinoura, N., et al. And Hamada. H: "Adenovirus-mediated transfer of Bax with caspase-8 controlled by myelin basic protein promoter induces enhanced apoptosis in gliomas"Cancer Gene Ther. (in press). (1999)
• [Publications] Shibagaki,N., Hanada,K., Yamaguchi,S., Yamashita,H., Shimada,S.and Hamada,H.: "Functional analysis of CD82 in the early phase of T cell activation : roles in cell adhesion and signal transduction." Eur.J.Immunol.28(4). 1125-1133 (1998)
• [Publications] Kanai,F., Kawakami,T., Hamada,H., Sadata,A., Yoshida,Y., Tanaka,T., Ohashi,M., Tateishi,K., Shiratori,Y., and Omata,M.: "Adenovirus-mediated transduction of Escherichia coli uracil phosphoribosyltransferase gene sensitizes cancer cells to low concentration of 5-fluorouracil." Cancer Res.58. 1946-1951 (1998)
• [Publications] Shiratori,Y., Kanai,F., Hikiba,Y., Moriyama,H., Hamada,H., Matsumura,M., Tanaka,T., Lan,K-H., Ohashi,M., Naito,M., and Omata,M.: "Gene therapy for hepatic micrometastasis of murine colon carcinoma." J.Hepatology. 28(5). 886-895 (1998)
• [Publications] Ohashi,M., Kanai,F., Tanaka,T., Lan,K-H., Shiratori,Y., Komatsu,Y., Kawabe,T., Yoshida,H., Hamada,H., and Omata,M.: "In vivo adenovirus-mediated prodrug gene therapy for carcinoembryonic antigen-pro-ducing pancreatic cancer." Jp.J.Cancer Res.89. 457-462 (1998)
• [Publications] Cao,X., Ju,D.W., Tao,Q., Wang,J., Wan,T., Wang,B.M., Zhang,W., and Hamada,H.: "Adenovirus-mediated GM-CSF gene and cytosine deaminase gene transfer followed by 5-fluorocytosine administration elicit more potent antitumor response in tumor-bearing mice." Gene Therapy. 5. 1130-1136 (1998)
• [Publications] Nakazaki Y, Tani K, Lin Z-T, Sumimoto H, Hibino H, Izawa K, Hase H, Takahashi S, Tojo A, Azuma M, Hamada H, Mori S, and Asano S.: "Vaccine effect of granulocyte-macrophage colony-stimulating factor or CD80 gene-transduced murine hematopoietic tumor cells and their co-operative en-hancement of antitumor immunity" Gene Therapy. 5(10). 1355-1362 (1998)
• [Publications] Shiniura,N., Yoshida,Y., Sadata,A., Hanada,K., Yamamoto,S., Kirino,T., Asai,A., and Hamada,H.: "Apoptosis by retrovirus-and adenovirus-mediated gene transfer of Fas ligand to glioma cells : implication for gene therapy." Human Gene Therapy. 9(14). 1983-1993 (1998)
• [Publications] Cao,X., Zhang,W., He,L., Xie,Z., Ma,S., Tao,Q., Yu,Y., Hamada,H.and Wang,J.: "Lymphotactin gene-modified bone marrow dendritic cells act as more potent adjuvants for peptide delivery to induce antitumor immunity." J.Immunol.161(11). 6238-6244 (1998)
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• [Publications] Yasuda H, Nagata M, Arisawa K, Yoshida R, Fujihira K, Okamoto N, Moriyama H, Miki M, Saito I, Hamada, H., Yokono K, Kasuga M.: "Local expression of immunoregulatory IL 12p40 gene prolonged syngeneic islet graft survival in diabetic NOD mice." J.Clin.Invest.102(10). 1807-1814 (1998)
• [Publications] Nagasaka T, Hayashi S, Tachi Y, Liu D, koike C, Namii Y, Katayama A, Negita M, Kobayashi T, Hamada H, Yokoyama I, Takagi H.: "Inhibitory effect of alpha(1, 2)fucosyltransferase recombinant adenoviral vector on alpha Gal expression." Transplant Proc.30(7A). 3837-3838 (1998)
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• [Publications] Matsumoto,O., Sunamura,M., Shimamura,H., Kodama,T., Hashimoto,W., Kobari,M., Kato,K., Takeda,K., Yagita,H., Okumura,K., Hamada,H. and Matsuno,S.: "Adjuvant immunotherapy using genetically engineered interleukin 12 secreting fibroblasts pre-vents recurrence after surgical resection of established tumors in a murine adenocarcinoma model." Surgery. in press. (1999)
• [Publications] Ohashi,M., kanai,F., Ueno,H., Tanaka,T., Kawakami,T., koike,Y., Okenoue,T., Shiratori,Y., Hamada,H.and Omata,M.: "In vivo adenovirus-mediated p53 gene therapy for gastric carcinoma in vitro and in vivo." Gut. in press. (1999)
• [Publications] Inaba,M., Sawada,H., Sadata,A., and Hamada,H.: "In vitro circumvention of 5-fluorouracil resistance in human stomach cancer cells by adenovi-rus-mediated transduction of Escherichia coli uracil phophoribosyltransferase gene." Jp.J.Cancer Res.90(3)in press. (1999)
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• [Publications] Ichikawa,T., Tamiya,T., Adachi,Y., Ono,Y., Matsumoto,K., Furuta,T., Yoshida,Y., Hamada,H.and Ohmoto,T.: "in vivo efficacy and toxity of 5-fluorocytosine/cytosine deaminase gene therapy for malignant gliomas mediated by adenovirus." Cancer Gene Therapy. in press. (1999)
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