Study on the gene therapy using a new generation adenovirus vector (gutless adenovirus)

新一代腺病毒载体（无肠腺病毒）的基因治疗研究

• 批准号: 13670656
• 负责人: UCHINO Makoto
• 金额: $ 2.62万
• 依托单位: Kumamoto 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-13670656/
• 关键词: gene therapy; adenovirus vector; helper-dependent; muscular dystrophy; attachment receptor; dystrophin; ヘルパーウイルス依存型アデノウイルスベクター; ジストロフィン; CAR; mdxマウス; tropism; RGD

We constructed a helper virus dependent adenoviral vector (HDAV), which deleted all genomes coding viral proteins. It carries only the therapeutic gene instead. We constructed two kinds of HDAV. One carries full-length dystrophin and lacZ gene as a marker, the other carries full-length dystrophin and CAR, an attachment receptor for adenovirus. These HDAVs are called HDAVLacZ-dys and HDAVCAR-dys, respectively. When the neonatal mdx mouse skeletal muscles are inoculated with the HDAVLacZ-dys vector under the non-immunosuppressive condition, 8-week after the inoculation dystrophin expression could be detected efficiently. We could confirm not only the dystrophin expression but also the normalization of the pathological abnormality in the transfected muscle cells. When HDAVCAR-dys vector was inoculated repetitively in to the maturated mdx mouse skeletal muscles, the number of dystrophin positive muscle cells increased by this repetitive infection. And the expression of the dystrophin could prolong for long time. We propose that the gene therapy by these strategies will be applied for many kinds of the muscular dystrophy.

我们构建了一个依赖辅助病毒的腺病毒载体（HDAV），它删除了所有编码病毒蛋白的基因组。它只携带治疗基因。我们构建了两种HDAV。一个携带全长肌营养不良蛋白和lacZ基因作为标记，另一个携带全长肌营养不良蛋白和CAR，一种腺病毒的附着受体。这些HDAV分别被称为HDAVLacZ-dys和HDAVCAR-dys。当在非免疫抑制条件下用HDAVLacZ-dys载体接种新生mdx小鼠骨骼肌时，接种后8周可以有效地检测肌营养不良蛋白表达。我们不仅可以确认抗肌萎缩蛋白的表达，而且还可以确认转染的肌肉细胞中病理异常的正常化。当HDAVCAR-dys载体重复接种到成熟的mdx小鼠骨骼肌中时，抗肌萎缩蛋白阳性肌细胞的数量通过这种重复感染而增加。抗肌萎缩蛋白的表达可持续较长时间。我们认为，这些策略的基因治疗将应用于多种类型的肌营养不良症。

项目成果

Maeda Y., Uchino M.et al.: "Cve/Lox P-mediated adenovirus type 5 packaging signal excision demonstrates that core-element VI is sufficient for virus packaging"Virology. (in press). (2003)

Maeda Y.、Uchino M.等人：“Cve/Lox P 介导的腺病毒 5 型包装信号切除表明核心元件 VI 足以进行病毒包装”病毒学。

Yamashita S., Uchino M.et al.: "Effect on motor neuron survival in mutant SOD1 (G93A) transgenic mice by BCl-2 expression using retrograde axonal transport of adenoviral vectors"Neurosci Letter. 328. 289-293 (2002)

Yamashita S.、Uchino M.等人：“使用腺病毒载体逆行轴突运输表达 BCl-2 对突变型 SOD1 (G93A) 转基因小鼠运动神经元存活的影响”Neurosci Letter。

Nishida Y., Maeda Y., Hara A., Arima T., Kimura E., Yamashita S., Uyama E., Mita S., Uchino M.: "Adenovirus-mediated murine interferon-g receptor transfer enhances the efficiency of IFN-g in vivo"Biochem. Biophys. Res. Commun.. 290. 1042-1047 (2002)

Nishida Y.、Maeda Y.、Hara A.、Arima T.、Kimura E.、Yamashita S.、Uyama E.、Mita S.、Uchino M.：“腺病毒介导的鼠干扰素 g 受体转移增强了

Kimura E., Uchino M et al.: "Efficient repetitive gene delivery to skeletal muscle using recombinant adenovirus vector containing the Coxsackievirus and Adenovirus receptor cDNA"Gene Therapy. 8. 20-27 (2001)

Kimura E.、Uchino M 等人：“使用含有柯萨奇病毒和腺病毒受体 cDNA 的重组腺病毒载体，将基因有效重复递送至骨骼肌”基因治疗。