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: 12217123
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Sapporo Medical University
• Principal Investigator: 濱田 洋文 札幌医科大学, 医学部, 教授 (00189614)
• Co-Investigator(Kenkyū-buntansha): 加藤 和則 札幌医科大学, 医学部, 助教授 (60233780) ; 中村 公則 札幌医科大学, 医学部, 講師 (80381276) ; 本望 修 札幌医科大学, 講師 (90285007) ; 伊藤 克礼 札幌医科大学, 医学部, 助手 (80332914) ; 佐々木 勝則 札幌医科大学, 医学部, 助手 (60336394)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥94,800,000 (Direct Cost: ¥94,800,000); Fiscal Year 2004: ¥19,300,000 (Direct Cost: ¥19,300,000); Fiscal Year 2003: ¥19,300,000 (Direct Cost: ¥19,300,000); Fiscal Year 2002: ¥19,000,000 (Direct Cost: ¥19,000,000); Fiscal Year 2001: ¥19,200,000 (Direct Cost: ¥19,200,000); Fiscal Year 2000: ¥18,000,000 (Direct Cost: ¥18,000,000)
• Keywords: 膵癌 / 悪性黒色腫 / 悪性神経膠腫 / B細胞系の腫瘍 / 前立腺癌 / モノクローナル抗体 / 遺伝子治療 / アデノウルスベクター / アデノウイルス / ファイバー変異型 / がん治療 / 分子標的 / モノクローン抗体 / NG2 / 口腔癌 / 膀胱癌 / 安全性評価 / アデノウイルスベクター / 悪性脳腫瘍 / 受容体 / ペプチドリガンド / がん / ファイバー

Research Abstract

腫瘍に対する選択的標的化の候補分子を探索するために、抗体のFcドメインに結合するProtein AのZ33モチーフをAd5ファイバーのHIループに持つAdv-FZ33アデノウイルスを作成した。CARをほとんど発現しないヒト膵癌細胞AsPc1やヒトメラノーマ細胞A375に対する遺伝子導入効率をEGFPないしb-gal遺伝子発現で測定した。AsPc1やA375に発現する表面分子(CD29、CD54など)に対する抗体を付着させたAdv-FZ33による遺伝子導入・遺伝子発現は、コントロール(抗体の非存在下またはコントロールIgG併用でのAdv-FZ33、ならびに野生型Ad5ファイバーAdv-Fwtのウイルス)による遺伝子導入・発現の数十倍に増強できた。また、ErbB2を高発現するヒト卵巣癌細胞(SK-OV3など)への遺伝子導入は、ErbB2抗体の併用により、選択的に著明に(EGFP遺伝子導入細胞%で、5%から90%へ)増強できた。腫瘍細胞とZ33アデノウイルスとを架橋することによって遺伝子導入効率が高まるモノクローナル抗体をスクリーニングすることにより、腫瘍細胞に対して標的化の可能な表面分子と抗体の組み合わせの探索を開始している。先行しているヒト膵癌を標的化できる新規抗体作製のプロジェクトでは、すでに4種類の抗体産生ハイブリドーマが樹立されている。このうちの一つクローンA^*によって得られるモノクローナル抗体は、AdvFZ33アデノウイルスの膵癌細胞AsPc1への遺伝子導入効率を、非常に強く(200倍)増強し、有望である。

Research Products

• [Journal Article] Mesenchymal stem cells that produce neurotrophic factors reduce ischemic damage in the rat middle cerebral artery occlusion model. (2005)
  • Author(s): Kurozumi K.et al., Hamada H.
  • Journal Title: Mol.Ther. 11(1) Pages: 96-104
• [Journal Article] Matsunaga T and Niitsu Y. Mesenchymal stem cells (MSC) as therapeutic cytoreagents for gene therapy. (2005)
  • Author(s): Hamada H. et al.
  • Journal Title: Cancer Science, (Review) (in press)
• [Journal Article] Bcl-xL transfer inhibits Bax translocation and prologs cardiac cold preservation time in rat. (2005)
  • Author(s): Huang J.et al., Hamada H.
  • Journal Title: Circulation (in press)
• [Journal Article] BDNF gene-modified mesenchymal stem cells promote functional recovery and reduce infarct size in the rat middle cerebral artery occlusion model. (2004)
  • Author(s): Kurozumi K.et al., Hamada H.
  • Journal Title: Mol.Ther. 9(2) Pages: 189-197
• [Journal Article] Anti-tumor effect of genetically engineered mesenchymal stem cells in a rat glioma model. (2004)
  • Author(s): Kurozumi K.et al., Hamada H.
  • Journal Title: Gene Ther. 11(14) Pages: 1155-1164
• [Journal Article] Adenovirus-Mediated Transfer of siRNA against Survivin Induced Apoptosis and Attenuated Tumor Cell Growth in Vitro and in Vivo. (2004)
  • Author(s): Uchida H.et al., Hamada H.
  • Journal Title: Mol.Ther. 10(1) Pages: 162-171
• [Publications] Kawano Y, et al. Hamada H.: "Ex vivo expansion of human umbilical cord hematopoietic progenitor cells using a coculture system with human telomerase catalytic subunit (hTERT)-transfected human stromal cells."Blood. 101(2). 532-540 (2003)
• [Publications] Uchida H.et al., Hamada H.: "5-Fluorouracil efficiently enhanced apoptosis induced by adenovirus-mediated transfer of caspase-8 in DLD-1 colon cancer cells."J.Gene Med.. 5(4). 287-299 (2003)
• [Publications] Nakamura T., Sato K., Hamada, H.: "Reduction of natural adenovirus tropism to the liver by both ablation of fiber-Coxsackievirus and adenovirus receptor interaction and use of replaceable short fiber."J.Virol.. 77(4). 2512-2521 (2003)
• [Publications] Dehari H, et al., Hamada H.: "Enhanced antitumor effect of RGD fiber-modified adenovirus for gene therapy of oral cancer."Cancer Gene Therapy. 10(1). 75-85 (2003)
• [Publications] Nakamura K.et al., Hamada H.: "Anti-tumor effect of genetically engineered mesenchymal stem cells in a rat glioma model."Gene Therapy. In press. (2004)
• [Publications] Nakamura T, Sato K., Hamada H.: "Effective Gene Transfer to Human Melanomas via Integrin-Targeted Adenoviral Vectors"Hum. Gene Ther.. 13(5). 613-626 (2002)
• [Publications] Kawano Y, et al., Hamada H.: "Ex vivo expansion of human umbilical cord hematopoietic progenitor cells using a coculture system with human telomerase catalytic subnit (hTERT)-transfected human stromal cells"Blood. 101(2). 532-540 (2003)
• [Publications] Uchida H, Shinoura N, Kitayama J, Watanabe T, Nagawa H, Hamada H.: "5-Fluorouracil efficiently enhanced apoptosis induced by adenovirus-mediated transfer of caspase-8 in DLD-1 colon cancer cells"J Gene Med..
• [Publications] Tsuda H, et al., Hamada H.: "Efficient BMP2 gene transfer and bone formation of mesenchymal stem cells by a fiber-mutant adenoviral vector"Mol. Ther.. 7(2). 1-12 (2003)
• [Publications] Nakamura T, Sato K, Hamada H.: "Reduction of natural adenovirus tropism to the liver by both ablation of fiber-Coxsackievirus and adenovirus receptor interaction and use of replaceable short fiber"J Virol.. 77(4). 2512-2521 (2003)
• [Publications] Dehari H, Ito Y, Nakamura T, Kobune M, Sasaki K, Yonekura N, Kohama G, Hamada H.: "Enhanced antitumor effect of RGD fiber-modified adenovirus for gene therapy of oral cancer"Cancer Gene Therapy. 10(1). 75-85 (2003)
• [Publications] Nakamura T, Sato K., Hamada, H.: "Effective gene transfer for human melanomas by integrin-targeted adenoviral vectors. Hum.Gene Ther.13(5):in press,2002"Human Gene Ther.. 13(5)(In press). (2002)
• [Publications] Yamamoto S., et al., Hamada, H.: "Reduced transduction efficiency of adenoviral vectors expressing human p53 gene by repeated transduction into glioma cells in vitro"Clinical Cancer Res.. (In press). (2002)
• [Publications] Uchida H, Shinoura N, Kitayama J, Watanabe T, Nagawa H, Hamada, H.: "Caspase-8 gene transduction augments radiation-induced apoptosis in DLD-1 cells. Biochem.Biophys.Res.Commun.in press,2002"Biochem.Biophys.Res.Commun. (In press). (2002)
• [Publications] Shinoura N, Furitsu T, Asai A, Kirino T, Hamada, H.: "Co-transduction of p27Kip1 strongly augments Fas ligand-and caspase-8-mediated apoptosis in U-373MG glioma cells"Anticancer Res.. 21(5). 3261-3268 (2001)
• [Publications] Shinoura N, Sakurai S, Asai A, Kirino T, Hamada, H.: "Caspase-9 transduction overrides the resistance mechanism against p53-mediated apoptosis in U-87MG glioma cells"Neurosurgery. 49(1):. 177-86 (2001)
• [Publications] Shinoura N, Sakurai S, Asai A, Kirino T, Hamada, H.: "Over-expression of APAF-1 and caspase-9 augments radiation-induced apoptosis in U-373MG glioma cells"Int J Cancer. 93(2). 252-61 (2001)
• [Publications] Motoi,F., et al. and Hamada,H.,: "Effective gene therapy for pancreatic cancer by cytokines mediated by restricted replication-competent adenovirus."Human Gene Ther.. 11. 223-235 (2000)
• [Publications] Shinoura,N., et al. and Hamada,H.,: "Adenovirus-mediated transfer of Bcl-xl protects neuronal cells from Bax-induced apoptosis."Exp.Cell Res.. 254. 221-231 (2000)
• [Publications] Shinoura,N., et al. and Hamada.H.,: "Adenovirus-mediated transfer of caspase-3 with Fas ligand Induces drastic apoptosis In U373MG glioma cells."Exp.Cell Res.. 256. 423-433 (2000)
• [Publications] Shinoura,N., et al.and Hamada,H.,: "Adenovirus-mediated transfer of caspase-8 augments cell death in gliomas : Implication for gene therapy."Human Gene Ther.. 11. 1123-1137 (2000)
• [Publications] Shinoura,N., et al.and Hamada,H.,: "Adenovirus-mediated transfer of caspase-8 in combination with super-repressor of NF-kB drastically induced apoptosis in gliomas."Biochem.Biophys.Res.Commun.. 271. 544-552 (2000)
• [Publications] Shinoura,N., et al.and Hamada,H.,: "Transduction of Apaf-1 or caspase-9 induces apoptosis in A172 cells, which are resistant to p53-mediated apoptosis."Biochem.Biophys.Res.Commun.. 272. 667-673 (2000)