癌への特異的標的化を可能とするアデノウイルスベクターシステムの開発

• Project/Area Number: 13218142
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: National Institute of Health Sciences
• Principal Investigator: 水口 裕之 国立医薬品食品衛生研究所, 基盤研究第三プロジェクトチーム, 副プロジェクト長 (50311387)
• Co-Investigator(Kenkyū-buntansha): 川端 健二 国立医薬品食品衛生研究所, 基盤研究第三プロジェクトチーム, 主任研究官 (50356234) ; 櫻井 文教 国立医薬品食品衛生研究所, 基盤研究第三プロジェクトチーム, 研究員 (70370939)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥40,600,000 (Direct Cost: ¥40,600,000); Fiscal Year 2004: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 2003: ¥10,400,000 (Direct Cost: ¥10,400,000); Fiscal Year 2002: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 2001: ¥10,200,000 (Direct Cost: ¥10,200,000)
• Keywords: 遺伝子治療 / アデノウイルスベクター / ファイバー / ターゲティング / 癌 / ターゲッティング / ターゲッテイング

Research Abstract

組織特異性を有したアデノウイルスベクターの開発には、nativeのアデノウイルスの受容体であるCAR(coxsackievirus-adenovirus receptor :第1の受容体)やαvインテグリン(第2の受容体)、ヘパラン硫酸(第3の受容体)を認識せず、ファイバーノブのHIループやC末端コード領域などに挿入した外来ペプチドを介して細胞特異的受容体を認識してのみ感染するベクターシステムの開発が必要である。本年度は、昨年度までに基本ユニットを開発済みのファイバーノブのFGループとペントンベース、ファイバーシャフトの3領域を同時に改変することで、上記の3つの受容体とは結合しないトリプルミュータントアデノウイルスベクターの改良を行った。CARとの結合能を除去するためのファイバーノブの変異として、FGループに変異(アミノ酸欠損)をもったベクター(昨年度に開発済み)に加え、ABループに変異(アミノ酸変異)を有したトリプルミュータントアデノウイルスベクター(ペントンベースのRGDモチーフの欠損、およびファイバーシャフト領域を35型アデノウイルス由来に置換)を作製した。これらのベクターの遺伝子導入・発現特性を検討したところ、ABループに変異を加えたベクターでは更なる遺伝子発現能の減弱が認められ、in vitroとin vivo(マウス)の両条件化でmock群に近いレベルの活性しか認められなかった。従って、特定の臓器で目的遺伝子の発現を起こさないアデノウイルスベクターの更なる改良に成功し、ターゲティングアデノウイルスベクターのための基盤ベクターになりうるものと期待された。

Research Products

• [Journal Article] Augmentation of the migratory ability of DC-based vaccine into regionalymph nodes by efficient CCR7 gene transduction. (2005)
  • Author(s): Okada, N., et al.
  • Journal Title: Gene Ther. 12 Pages: 129-139
• [Journal Article] Immunological properties and vaccine efficacy of murine dendritic cells simultaneously expressing melanoma-associated antigen and interleukin-12. (2005)
  • Author(s): Okada, N., et al.
  • Journal Title: Cancer Gene Ther. 12 Pages: 72-83
• [Journal Article] Targeted adenovirus vectors (2004)
  • Author(s): Mizuguchi, H. et al.
  • Journal Title: Hum. Gene Ther. 15 Pages: 1022-1033
• [Journal Article] Adenovirus vector-mediated doxycycline-inducible RNA interference. (2004)
  • Author(s): Hosono, T. et al.
  • Journal Title: Hum. Gene Ther. 15 Pages: 813-819
• [Journal Article] RNA interference of PPARγ using fiber-modified adenovirus vector efficiently suppresses preadipocyte-to-adipocyte differentiation in 3T3-L1 cells.
  • Author(s): Hosono, T., et al.
  • Journal Title: Gene (In press)
• [Journal Article] Transcriptional targeting of RGD fiber-mutant adenovirus vectors can improve the safety of suicide gene therapy for murine melanoma.
  • Author(s): Okada, Y., et al.
  • Journal Title: Cancer Gene Ther. (In press)
• [Publications] Okada Y., Okada N., Mizuguchi H., Takahashi K., Hayakawa T., Mayumi T., Mizuno N.: "Optimization of antitumor efficacy and safety of in vivo cytokine gene therapy using RGD fiber-mutant adenovirus vector for preexisting murine melanoma."Blochim.Biophys.Acta.. 1670. 172-180 (2004)
• [Publications] Koizumi N., Mizuguchi H., Sakurai F., Yamaguchi T., Watanabe Y., Hayakawa T.: "Reduction of natural adenovirus tropism to mouse liver by fiber-shaft exchange in combination with both CAR- and αv integrin-binding ablation."J.Virol.. 77. 13062-13072 (2003)
• [Publications] Gao J-Q., Tsuda Y., Katayama K., Nakayama T., Hatanaka Y., Tani Y., Mizuguchi H., Hayakawa T., Yoshie O., Tsutsumi Y., Mayumi T., Nakagawa S.: "Antitumor effect by interleukin-11 receptor α-locus chemokine/CCL27, introduced into tumor cells through a recombinant adenovirus vector."Cancer Res.. 63. 4420-4425 (2003)
• [Publications] Koizumi N., Mizuguchi H., Utoguchi N., Watanabe Y., Hayakawa T.: "Generation of fiber-modified adenovirus vector containing heterologous peptides in both the HI loop and C terminus of the fiber knob."J.Gene Med. 5. 267-276 (2003)
• [Publications] Nagayama Y., Nakao, K., Mizuguchi H., Hayakawa T., Niwa M.: "Enhanced antitumor effect of combined replicative adenovirus and non-replicative adenovirus expressing interleukin-12 in an immunocompetent mouse model."Gene Ther.. 10. 1400-1403 (2003)
• [Publications] Okada Y., Okada N., Mizuguchi H., Hayakawa T., Mayumi T., Mizuno N.: "An investigation of adverse effects caused by the injection of high-dose TNFα-expressing adenovirus vector into established murine melanoma."Gene Ther.. 10. 1891-1902 (2003)
• [Publications] Okada N., Masunaga Y., Okada Y., Iiyama S., Tsuda T., Matsubara A., Mori N., Mizuguchi H., Hayakawa T., Fujita T., Yamamoto A.: "Gene transduction efficiency and the state of maturation in mouse dendritic cells Infected with conventional or RGD fiber-mutant adenovirus vectors."Cancer Gene Ther.. 10. 421-431 (2003)
• [Publications] Kakayama K., Wada K., Nakajima A., Yoshida S., Mizuguchi H., Hayakawa T., Nakagawa S., Kadowaki T., Nagai R., Kamisaki Y., Blumberg RS., Mayumi T.: "A novel PPARγ-gene therapy to control inflammation associated with inflammatory bowel disease."Gastroenterology. 124. 1315-1324 (2003)
• [Publications] Koizumi N., Mizuguchi H., Utoguchi N., Watanabe Y., Hayakawa T.: "Generation of fiber-modified adenovirus vector containing heterologous peptides in both the HI loop and C terminus of the fiber knob"J.Gene Med. (印刷中). (2003)
• [Publications] Okada Y., Okada N., Mizuguchi H., Hayakawa T., Mayumi T., Mizuno N.: "An investigation of adverse effects caused by the injection of high-dose TNFα-expressing adenovirus vector into established murine melanoma"Gene Ther.. (印刷中). (2003)
• [Publications] Okada N., Masunaga Y., Okada Y., Iiyama S., TsudaT., Matsubara A., Mon N., Mizuguchi H., Hayakawa T., Fujita T., Yamamoto A.: "Gene transduction efficiency and the state of maturation in mouse dendritic cells infected with conventional or RGD fiber-mutant adenovirus vectors"Cancer Gene Ther.. (印刷中). (2003)
• [Publications] Mizuguchi H., Koizumi N., Hosono T., Ishii-Watabe A., Uchida E., Utoguchi N., Watanabe Y., Hayakawa T.: "CAR-or αv integrin-binding ablated adenovirus vectors, but not fiber-modified vectors containing RGD peptide, do not change the systemic gene transfer properties in mice"Gene Ther.. 9. 769-776 (2002)
• [Publications] Mizuguchi H., Hayakawa T.: "Enhanced anti-tumor effect and reduced vector dissemination with fiber-modified adenovirus vectors expressing herpes simplex virus thymidine kinase"Cancer Gene Ther.. 9. 236-242 (2002)
• [Publications] Okada Y., Okada N., Nakagawa S., Mizuguchi H., Takahashi K., Mizuno N., Fujita T., Yamamoto A., Hayakawa T., Mavumi T.: "Tumor necrosis factor a-gene therapy for an established murine melanoma using RGD (Arg-Gly-Asp) fiber-mutant adenovirus vectors"Jpn.J.Cancer Res.. 93. 436-444 (2002)
• [Publications] H. Mizuguchi, N.Koizumi, T.Hosono, N. Utoguchi, Y.Watanabe, M.A.Kay, T.Hayakawa.: "A simplified system for constructing recombinant adenoviral vectors containing heterologous peptides in the HI loop of their fiber knob"Gene Ther.. 8. 730-735 (2001)
• [Publications] N.Okada, Y.Tsukada, S.Nakagawa, H.Mizuguchi, K.Mori, T.Saito, T.Fujita, A.Yamamoto, T.Hayakawa, T.Mayumi.: "Efficient gene delivery into dendritic cells by fiber-mutant adenovirus vectors"Biochem.Biophys.Res.Commun.. 282. 173-179 (2001)
• [Publications] H.Mizuguchi, T.Hayakawa.: "Approaches for generating recombinant adenovirus vectors"Adv.Drug.Deli.Rev.. 52. 165-176 (2001)
• [Publications] H.Mizuguchi, T.Hayakawa.: "Enhanced anti-tumor effect and reduced vector dissemination with fiber-modified adenovirus vectors expressing herpes simplex virus thymidine kinase"Cancer Gene Ther.. (in press). (2002)
• [Publications] N.Koizumi, H.Mizuguchi, T.Hosono, A.Ishii-Watabe, E.Uchida, N.Utoguchi, Y.Watanabe, T.Hayakawa.: "Efficient gene transfer by fiber-mutant adenoviral vectors containing RGD peptide"Biochim.Biophys.Acta. 1568. 13-20 (2001)
• [Publications] N.Okada, T.Saito, Y.Masunaga, Y.Tsukada, S.Nakagawa, H.Mizuguchi, K.Mori, Y.Okada, T.Fujita, T.Hayakawa, T.Mayumi, A.Yamamoto.: "Efficient antigen gene transduction using Arg-Gly-Asp fiber-mutant adenovirus vectors can potentiate anti-tumor vaccine efficacy and maturation of murine dendritic cells"Cancer Res.. 61. 7913-7919 (2001)