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

我们构建了一个辅助病毒依赖腺病毒载体（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腺病毒附着受体CAR。这些hdav分别被称为HDAVLacZ-dys和HDAVCAR-dys。将HDAVLacZ-dys载体在非免疫抑制条件下接种于新生mdx小鼠骨骼肌，接种8周后可有效检测肌营养不良蛋白的表达。我们不仅证实了肌营养不良蛋白的表达，而且证实了肌细胞病理异常的正常化。将HDAVCAR-dys载体反复接种于成熟的mdx小鼠骨骼肌中，这种反复感染使肌营养不良蛋白阳性细胞的数量增加。抗肌营养不良蛋白的表达可延长较长时间。我们认为这些策略的基因治疗将应用于多种类型的肌肉萎缩症。

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.