Basic Research on gene transfer using gene-gun against malignant gliomas

基因枪抗恶性胶质瘤的基因转移基础研究

• 批准号: 10470285
• 负责人: ASAI Akio
• 金额: $ 7.94万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470285/
• 关键词: gene therapy; gene-gun; glioma; adenovirus

(1) Ex vivo gene transfer with gene-gunWe evaluated gene-gun-mediated ex vivo gene transfer using subcuateous tumor-bearing rat model. First we transplanted 9L glioma cells into F344 syngeneic rats subcutaneously and surgically removed formed tumors 20 days after transplantation. Lac Z expression vector was transduced by gene-guun at shooting pressure (psi) 100, 200, 400, and 600 (max) or Lca Z expressing adenovirus was incubated with the tumor at MOI 20, 40, and 100 for 24 hours. The tumors were then re-transplanted subcutaneously and gene expression was evaluated 7 days after the re-transplantation. The highest gene-gun-mediated gene transfer rate ranged from 3 to 9 % along with shooting pressure whereas that by adenovirus ranged from 5.5 to 15.9% along with MOI which are statistically different.(2) In vivo gene transfer with gene-gunNext, we estimated gene-gun mediated in vivo gene transfer using the same animal model as (1). We surgically opened the skin over the subcutaneously formed tumors and shot Lac Z expression vector at the same psi as (1) or injected Lac Z expression adenovirus at the same MOIs as (1). After the treatments, the wounds were closed and gene expression was examined 7 days after the treatments. The highest gene-gun-mediated gene transfer rate ranged from 3.6 to 9.5 % along with shooting pressure whereas that by adenovirus ranged from 5.8 to 13.2 % along with MOI which are statistically different.In conclusion, although gene transfer with gene-gun is less effective than adenovirus-mediated gene transfer against gliomas, gene-gun-mediated gene transfer is still a good tool to transduce genes in vivo or ex vivo.

（1）使用基因 - 冈韦的离体基因转移使用亚肿瘤下肿瘤大鼠模型评估了基因-Gun介导的离体基因转移。首先，我们将9L神经胶质瘤细胞皮下移植到F344合成大鼠中，并在移植后20天手术切除了形成的肿瘤。 LAC Z表达载体在射击压力（PSI）100、200、400和600（MAX）或表达腺病毒的LCA Z时通过Gene-guun转导，在MOI 20、40和100中孵育24小时。然后对肿瘤进行皮下重新移植，并在重新移植后7天评估基因表达。最高的基因-GUN介导的基因转移率范围为3至9％，而腺病毒的腺病毒范围为5.5至15.9％，而MOI则在统计学上不同。（2）带有基因基因转移的基因转移。我们使用与（1）相同的动物模型估计基因枪介导的体内基因转移。我们在皮下形成的肿瘤上手术打开皮肤，并在与（1）或注射的lac z表达腺病毒相同的MOI与（1）相同的lac z表达腺病毒的PSI上拍摄Lac Z表达载体。治疗后，闭合伤口并在治疗后7天检查基因表达。最高的基因-GUN介导的基因转移率范围为3.6至9.5％，而拍摄压力的范围为5.8至13.2％，而MOI则在统计学上不同。比腺病毒介导的基因转移对神经胶质瘤的有效，基因辅助介导的基因转移仍然是在体内或体内转导基因的好工具。

项目成果

Chi S, Kitanaka C, Noguchi K, Mochizuki T, Nagashima Y, Shirouzu M, Fujita H, Yoshida M, Chen W, Asai A, Himeno M, Yokoyama S, Kuchino Y: "Oncogenic Ras trigger cell suicide through the activation of a caspase-independent cell death program in human cance

Chi S、Kitanaka C、Noguchi K、Mochizuki T、Nagashima Y、Shirouzu M、Fujita H、Yoshida M、Chen W、Asai A、Himeno M、Yokoyama S、Kuchino Y：“致癌 Ras 通过激活

Shinoura N, Yoshida Y, Tsunoda R, Ohashi M, Zhang W, Asai A, Kihno T, Hamada H: "Highly augmented cytopathic effect of a fiber-mutant E1B-defective adenovirus for gene therapy of gliomas."Cancer Res. 59. 3411-3416 (1999)

Shinoura N、Yoshida Y、Tsunoda R、Ohashi M、Zhang W、Asai A、Kihno T、Hamada H：“纤维突变型 E1B 缺陷型腺病毒对神经胶质瘤基因治疗的高度增强的细胞病变作用。”Cancer Res。

Chi S, Kitanaka C, Noguchi K, Mochizuki T, Nagashima Y, Shirouzu M, Fujita H, Yoshida M, Chen W, Asai A, Himeno M, Yokoyama S, Kuchino: "Oncogenic Ras triggers cell suicide through the activation of a caspase-independent cell death program in human cancer

Chi S、Kitanaka C、Noguchi K、Mochizuki T、Nagashima Y、Shirouzu M、Fujita H、Yoshida M、Chen W、Asai A、Himeno M、Yokoyama S、Kuchino：“致癌 Ras 通过激活 caspase 触发细胞自杀

Nagane M, Asai A, Shibui S, Oyama H, Nomura K, Kuchino Y: "Expression pattern of chemoresistance-related genes in human malignant brain tumors: a working knowledge for proper selection of anticancer drugs"Jpn J Clin Oncol. 29. 527-534 (1999)

Nagane M、Asai A、Shibui S、Oyama H、Nomura K、Kuchino Y：“人类恶性脑肿瘤中化疗耐药相关基因的表达模式：正确选择抗癌药物的工作知识”Jpn J Clin Oncol。

Nagane M, Asai A, Shibui S, Oyama H, Nomura K, Kuchino Y: "Expression pattern of chemoresistance-related genes in human malignant brain tumors : a working knowledge for proper selection of anticancer drugs."Jpn J Clin Oncol. 29. 527-534 (1999)

Nagane M、Asai A、Shibui S、Oyama H、Nomura K、Kuchino Y：“人类恶性脑肿瘤中化疗耐药相关基因的表达模式：正确选择抗癌药物的工作知识。”Jpn J Clin Oncol。