Development of Fiber-Mutant Adenovirus Vectors for targeting gene therapy

用于靶向基因治疗的纤维突变腺病毒载体的开发

• 批准号: 13672282
• 负责人: NAKAGAWA Shinsaku
• 金额: $ 2.3万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672282/
• 关键词: gene therapy; adenovirus vectors; targeting; RGD; CAR; integrins; dendritic cells; phage libraries; メラノーマ細胞

We have demonstrated the usefulness of dendritic cells (DCs) genetically modified by adenovirus vectors (Ad) to immunotherapy, while sufficient gene transduction into DCs is required for high doses of Ad. The RT-PCR analysis revealed that the relative resistance of DCs to Ad-mediated gene transfer is due to the absence of Coxsackie-adenovirus receptor expression, and that DCs expressed adequate alpha(v)-integrins. Therefore, we investigated whether fiber-mutant Ad containing the Arg-Gly-Asp (RGD) sequence in the fiber knob can efficiently transduce and express high levels of the LacZ gene into DCs. The gene delivery by fiber-mutant Ad was more efficient than that by conventional Ad in both murine DC lines and normal human DCs (NHDC). Furthermore, NHDC transduced with fiber-mutant Ad and conventional Ad at 8000-vector particles/cell resulted in a 70-fold difference in beta-galactosidase activity. We propose that alpha(v)-integrin-targeted Ad is a very powerful tool with which to implement DC-based vaccination strategies.

我们已经证明了由腺病毒载体（Ad）遗传修饰的树突状细胞（DC）对免疫治疗的有用性，而高剂量的Ad需要足够的基因转导到DC中。RT-PCR分析显示，DC对Ad介导的基因转移的相对抗性是由于缺乏柯萨奇-腺病毒受体表达，并且DC表达足够的α（v）-整合素。因此，我们研究了在纤维球中含有Arg-Gly-Asp（RGD）序列的纤维突变体Ad是否可以有效地将LacZ基因转染并高水平表达到DC中。在小鼠DC系和正常人DC（NHDC）中，纤维突变型Ad的基因递送比常规Ad更有效。此外，NHDC转导纤维突变的广告和传统的广告在8000-载体颗粒/细胞导致在β-半乳糖苷酶活性的70倍的差异。我们建议，α（V）-整合素靶向的广告是一个非常强大的工具，实现基于DC的疫苗接种策略。

项目成果

N.OKADA: "Efficient Antigen Gene Transduction Using Arg-Gly-Asp Fiber-Mutant Adenovirus Vectors Can Potentiate Antitumor Vaccine Efficacy and Maturation of Murine Dendritic Cells"Cancer Res.. 61・21. 7913-7919 (2001)

N.OKADA：“使用精氨酸-甘氨酸-天冬氨酸纤维突变腺病毒载体进行有效的抗原基因转导可以增强抗肿瘤疫苗的功效和鼠树突状细胞的成熟”Cancer Res.. 7913-7919 (2001)

S. Nakagawa et al.: "Tetracycline-regulatable adenovirus vectors: pharmacologic properties and clinical potential"Eur. J. Pharm. Sci.. 13. 53-60 (2001)

S. Nakakawa 等人：“四环素可调节腺病毒载体：药理学特性和临床潜力”Eur。

Y.Okada: "Tumor Necrosis Factor alfa-Gene Therapy for an Established Miurine Melanoma Using RGD (Arg-Gly-Asp) Fiber-mutant Adenovirus Vectors"Jpn. J. Cncer Res.. 93. 436-444 (2002)

Y.Okada：“使用 RGD (Arg-Gly-Asp) 纤维突变腺病毒载体对已确定的 Miurine 黑色素瘤进行肿瘤坏死因子 α 基因治疗”Jpn。

Y.Okada: "Tumor Necrosis Factor alfa-Gene Therapy for an Established Murine Melanoma Using RGD (Arg-Gly-Asp) Fiber-mutant Adenovirus Vectors"Jpn. J. Cncer Res.. 93. 436-444 (2002)

Y.Okada：“使用RGD（精氨酸-甘氨酸-天冬氨酸）纤维突变腺病毒载体对已建立的小鼠黑色素瘤进行肿瘤坏死因子α基因治疗”Jpn。

Y.Okada: "Fiber-mutant technique can augment gene transduction efficacy and anti-tumor effects against established murine melanoma by cytokine-gene therapy using adenovirus vectors"Cancer Letter. 1777. 57-63 (2002)

Y.Okada：“通过使用腺病毒载体进行细胞因子基因治疗，纤维突变技术可以增强基因转导功效和对已建立的小鼠黑色素瘤的抗肿瘤作用”《癌症信》。