ANTI-TUMOR EFFECT OF ANTI-ANGIOGENIC FACTOR GENES PRODUCED BY ELECTROPORATION

• Project/Area Number: 14571176
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Digestive surgery
• Research Institution: CHIBA UNIVERSITY
• Principal Investigator: GUNJI Yoshio CHIBA UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, ASSOCIATE PROFESSOR, 大学院・医学研究院, 助教授 (60241957)
• Co-Investigator(Kenkyū-buntansha): MIYAZAKI Shinichi CHIBA UNIVERSITY, UNIVERSITY HOSPITAL, ASSISTANT, 医学部附属病院, 助手 (40334198) ; SHIMADA Hideaki CHIBA UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, LECTURER, 大学院・医学研究院, 講師 (20292691) ; MATSUBARA Hisahiro CHIBA UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, LECTURER, 大学院・医学研究院, 講師 (20282486) ; OCHIAI Takenori CHIBA UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, PROFESSOR, 大学院・医学研究院, 教授 (80114255)
• Project Period (FY): 2002 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2003: ¥1,700,000 (Direct Cost: ¥1,700,000); Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
• Keywords: 電気穿孔法 / 遺伝子治療 / 血管新生抑制遺伝子 / 抗腫瘍効果 / 電撃化学療法 / electroporation / angiostatin / endostatin / immunotherapy / gene therapy

Research Abstract

The antitumor efficiency of electrochemotherapy using chemotherapeutic agents and high voltage electric pulse has been reported. This study was done to define the precise nature of the involvement of antitumor immunity in the regression of tumor nodules in electrochemotherapy, and to evaluate the effectiveness of using low voltage electroporation. Colon 26 tumors were eradicated in the mice given an intratumor (i.t.) injection of 500 μg bleomycin followed by treatment with electric fields ranging from 50 to 150V/cm, with complete response rates ranging from 80 to 100%. The mice rejected inoculations of re-challenged colon 26 cells, but not Meth A cells. In the Balb/c nu/nu nude mice, complete regression of the tumor was not seen after electrochemotherapy tinder the same therapeutic conditions that resulted in almost complete cure in the Balb/c mice. Our results suggest that the generation of T-cell-dependent, tumor-specific protective immunity might be involved in the process of tumor nodule regression in low-voltage electrochemotherapy.
Anti-angiogenic factors are potent endothelial cell growth inhibitors that have been shown to inhibit angiogenesis in vitro and tumor growth in mice. We have demonstrated the synergistic anti-tumor effect of anti-angiogenic genes (mouse angiostatin : pBLAST-mAngio, and mouse endostatin : p-BLAST42-mEndo XV) delivered to tumors by low-voltage electroporation in mouse colon 26 models. A synergistic anti-tumor effect was strongly suggested by in vivo tumor growth kinetics, as well as in survival studies with the mice. RT-PCR confirmed the fragments of each gene were transferred by low-voltage electroporation in the tumor. Decreased microvessel density measurements in tumors also confirmed the efficacy of the synergistic antitumor effect of both genes. Significant growth inhibition was observed in mice treated with a 1:1-proportion of angiostatin and endostatin genes, and the order of the both genes transferred (first the endostatin-gene, followed one week later by the angiostatin-gene) had a profound inhibitory effect on tumor growth. These data suggest that in viva delivery of anti-angiogenic genes with low-voltage electroporation could be a possible therapeutic strategy for established solid tumors when both genes were applied in combination.

Research Products

• [Publications] Miyazaki, S., Gunji.Y.et al.: "Possible invlvement of antitumor immunity in the mechanism of eradication of colon 26 induced by low-voltage electrochemotherapy with bleomycin."Surg Today. 33. 39-44 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Gunji, Y., Uesato, M.et al.: "Anti-tumor effect produced by low-voltage-electroporation-mediated transfer of angiostatin and ndostatin gene in mice."Molecular Therapy. 7. S414-S414 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Gunji, Y., et al.: "Analysis of the experimental liver metastasis from inoculated tumor in the mouse stomach cavity."Surg Today. 32. 142-147 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Gunji Y., Yol, S., Hayashi, H., Matsubara, H., Hori, S., Suzuki, T., Ochiai, T.: "Analysis of the experimental liver metastasis from inoculated tumor in the mouse stomach cavity"Sung Today. 32. 142-147 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Shimada, H., Matsubara, Y., Gunji, Y., Tagawa; M., Ochiai T.: "Gene therapy for esophageal cancer."Res.Adv.In cancer. 2. 45-52 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Gunji, Y., Uesato, M., Ochiai, T., Hideaki, S., Matsubara, H., Miyazaki, S., Nabeya, Y.: "Anti-tumor effect produced by low-voltage-electnoporation-mediated transfer of angiostatin and ndostatin gene in mice."6^<th> Annual meeting of the American Society of Gene Therapy. (Molecular Therapy 7, S414-S414). (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Miyazaki, S., Gunji, Y.et al.: "Possible invlvement of antitumor immunity in the mechanism of eradication of colon 26 induced by low-voltage electrochemotherapy with bleomycin."Surg Today. 33. 39-44 (2003)
• [Publications] Gunji, Y., Uesato, M.et al.: "Anti-tumor effect produced by low-voltage-electroporation-mediated transfer of angiostatin and ndostatin gene in mice."Molecular Therapy. 7. S414-S414 (2003)
• [Publications] 宮崎信一, 軍司祥雄 et al.: "Electrochemotherapyによる腫瘍退縮と宿主の免疫学的抗腫瘍効果の関与"日外会誌. 103. 542-542 (2002)
• [Publications] 上里昌也, 軍司祥雄 et al.: "大きな腫瘍に対するElectrochemotherapy繰り返し施行の効果"日外会誌. 103. 654-654 (2002)
• [Publications] Miyazaki, S., Gunji, Y., et al.: "Possible involvement of antitumor immunity in the mechanism of eradication of colon 26 induced by low-voltage electrochemotherapy with bleomycin"Surgery Today. (in press). (2003)