Experimental Efficacy of Genechemotherapy for Biliary cancer using molecular target gene transduction method.

• Project/Area Number: 21790648
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Gastroenterology
• Research Institution: University of Tsukuba
• Principal Investigator: FUKUDA Kuniaki 筑波大学, 医学医療系, 講師 (50447257)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2009: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 胆道学 / 遺伝子治療 / 消化器内科学

Research Abstract

Biliary cancers are highly malignant diseases with poor prognosis and thus development of new treatments such as gene therapy is necessary. Although adenovirus vectors are widely used for cancer gene therapy, conventional adenoviral vectors infect not only cancer cells but also normal cells. Therefore, a new method that allow more specific gene delivery to cancer cells needs to be developed to obtain more efficient anti-tumor effect and safety. We explored methods of retargeting adenovirus vectors for targeted gene therapy of human biliary cancers using the Ad incorporating an IgG Fc binding motif (Z33) from the Staphylococcus protein A (Ad-FZ33) combined with tumor specific antibodies. Flow cytometry analysis revealed high expression levels of epithelial cell adhesion molecule (EpCAM) and epidermal growth factor receptor (EGFR) on human biliary cancer cells. Ad-FZ33 expressing LacZ combined with antibodies against EpCAM or EGFR, followed by β-gal assay, demonstrated highly efficient gene transduction in these biliary cancer cells, compared with the treatment with control antibody or without antibody. Ad-FZ33 expressing uracil phosphoribosyl transferase (UPRT), an enzyme which greatly enhances the toxicity of 5-fluorouracil (FU), combined with antibodies against EpCAM or EGFR, remarkably enhanced the sensitivity of biliary cancer cells to 5-FU. By contrast, the treatment didn't affect 5-FU sensitivity of the cells not expressing EpCAM or EGFR including normal hepatocytes. Finally, treatments with the UPRT-expressing Ad-FZ33 with antibodies against EpCAM or EGFR, followed by 5-FU administration, were significantly suppressed the growth of biliary cancer xenografts in nude mice. These results indicate that the molecular target genechemotherapy mediated by the Z33 fiber-modified adenovirus with anti-EpCAM or anti-EGFR antibodies offers a potentially effective therapeutic modality against biliary cancers, and can be expected in clinical applications fairly.

Research Products

• [Journal Article] EpCAM-and EGFRtargetedselective gene therapy for biliarycancers using Z33-fiber modified adenovirus (2011)
  • Author(s): Kawashima R, Abei M, Fukuda K, NakamuraK, Murata T, Wakayama M, Seo E, Hasegawa N, Mizuguchi H, Obata Y, Hyodo I, Hamada H, Yokoyama KK
  • Journal Title: Int. J Cancer Volume: 129(5) Pages: 1244-1253
• [Journal Article] EpCAM- and EGFR-targeted selective gene therapy for biliary cancers using Z33-fiber modified adenovirus (2011)
  • Author(s): Kawashima R, Abei M, Fukuda K, Nakamura K, Murata T, Wakayama M, Seo E, Hasegawa N, Mizuguchi H, Obata Y, Hyodo I, Hamada H, Yokoyama KK
  • Journal Title: International Journal of Cancer Volume: 129 Pages: 1244-1253
  • Peer Reviewed
• [Journal Article] E1A, E1B double-restricted replicative adenovirus at low dose greatly augments tumor-specific suicide gene therapy for gallbladder cancer (2009)
  • Author(s): Fukuda K, Abei M, Ugai H, Kawashima R, Seo E, Wakayama M, Murata T, Endo S, Hamada H, Hyodo I, Yokoyama KK
  • Journal Title: Cancer Gene Ther Volume: 16(2) Pages: 126-36
• [Presentation] Targeted gene therapy for cancers by using Z33 fiber modified adenovirus vector with tumor specific antibodies (2009)
  • Author(s): Kawashima R, Fukuda K, Abei M, Nakamura K, Murata T, Yokoyama KK, Hamada H, Hyodo I
  • Organizer: 20th Asia Pacific Cancer Conference
  • Place of Presentation: つくば
  • Year and Date: 2009-11-12
• [Presentation] Targeted gene therapy for cancers by using Z33 fiber modified adenovirus vector with tumor specific antibodies (2009)
  • Author(s): Rei Kawashima
  • Organizer: 20^<th> Asia Pacific Cancer Conference
  • Place of Presentation: つくば市
  • Year and Date: 2009-11-12
• [Presentation] Z33アデノウイルスと腫瘍特異抗体を用いた胆道癌の標的化遺伝子治療の開発 (2009)
  • Author(s): 川島玲,安部井誠人,福田邦明,村田武英,濱田洋文,兵頭一之介
  • Organizer: 第45回日本胆道学会
  • Place of Presentation: 千葉
  • Year and Date: 2009-09-19
• [Presentation] Effective gene therapy for biliary cancers by using Z33 fiber modified adenovirus with tumor specific antibodies (2009)
  • Author(s): Kawashima R, Abei M, Fukuda K, Nakamura K, Murata T, Hyodo I, Hamada H, Obata Y, Yokoyama KK
  • Organizer: 第15回日本遺伝子治療学会
  • Place of Presentation: 大阪
  • Year and Date: 2009-07-10
• [Remarks]
  • URL: http://www.md.tsukuba.ac.jp/clinical-med/gastroenterology/daigakuin.html