Efficient repetitive gene delivery to skeletal muscle using recombinant adenovirus vector containing the Coxsackievirus and adenovirus receptor cDNA
Efficient repetitive gene delivery to skeletal muscle using recombinant adenovirus vector containing the Coxsackievirus and adenovirus receptor cDNA
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DOI：
10.1038/sj.gt.3301359
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发表时间：
2001-01-01
期刊：
影响因子：
5.1
通讯作者：
Uchino, M
中科院分区：
文献类型：
--
作者：
Kimura, E;Maeda, Y;Uchino, M
为了提高腺病毒介导的基因传递到骨骼肌，我们使用了重组腺病毒载体编码的人柯萨奇病毒和腺病毒受体（hCAR）。由于CAR在啮齿动物成肌细胞和肌纤维中的表达水平低于其他组织，因此我们预期CAR在骨骼肌中的表达升高将提高腺病毒介导的基因转移的功效。由于小鼠成肌细胞C2C12细胞对重组腺病毒5的感染表现出低敏感性，我们最初用含有hCAR cDNA和LacZ基因的重组腺病毒以250的高感染复数（MOI）感染这些细胞。随后用含有标记基因绿色荧光蛋白的重组腺病毒感染，即使在25的低MOI下也变得有效。因此，小鼠肌肉纤维中hCAR表达的升高使得以低剂量进行第二次病毒接种成为可能，我们还证明hCAR表达的升高不会影响肌肉膜的完整性。我们的研究结果表明，CAR和治疗基因的腺病毒载体的共表达构成了一个新的战略，以推进遗传性肌肉疾病的基因治疗。
To improve adenovirus-mediated gene delivery to skeletal muscle, we have used a recombinant adenovirus vector encoding the human Coxsackievirus and adenovirus receptor (hCAR). Because CAR is expressed at a lower level in rodent myoblasts and muscle fibers than in other tissues, we expected that elevated expression of CAR in skeletal muscle would improve the efficacy of adenovirus-mediated gene transfer. Since the mouse myoblasts, C2C12 cells, showed low sensitivity to infection by recombinant adenovirus 5, we initially infected these cells at a high multiplicity of infection (MOI) of 250 with the recombinant adenovirus containing hCAR cDNA and LacZ gene. Subsequent infection by recombinant adenovirus containing the marker gene, green fluorescence protein, became efficient even at a low MOI of 25. Thus, elevated hCAR expression in mouse muscle fibers made a second virus inoculation at low doses possible, We also demonstrated that the elevated hCAR expression did not influence muscle membrane integrity. Our results suggest that co-expression of CAR and a therapeutic gene by adenovirus vector constitutes a novel strategy to advance gene therapy for hereditary muscle diseases.