Wells Kim E, Torelli Silvia, Lu Qi, Brown Susan C, Partridge Terence, Muntoni Francesco, Wells Dominic J
Department of Neuromuscular Diseases, Imperial College Faculty of Medicine, Charing Cross Hospital, St Dunstan's Road, W6 8RP, London, UK.
Neuromuscul Disord. 2003 Jan;13(1):21-31. doi: 10.1016/s0960-8966(02)00191-8.
A lack of effective treatments for Duchenne muscular dystrophy, a fatal X-linked myopathy, has focused attention on the possibility of gene therapy. The aim of the gene therapy approach is the restoration of the dystrophin associated complex of proteins, one member of which is neuronal nitric oxide synthase, an important enzyme in signal transduction. Transgenic mdx mice and plasmid gene transfer of both human and murine recombinant dystrophins was used to assess whether nNOS could be restored to the sarcolemma following dystrophin gene transfer at a variety of levels of expression. Murine revertant fibres and human patients with different dystrophin deletions were used to assess the relationship between exon deletion and loss of neuronal nitric oxide synthase localization to the sarcolemma. We demonstrate that the domain encoded by exons 45-48 is required for localization of neuronal nitric oxide synthase to the sarcolemma. On the basis of these observations we suggest that neuronal nitric oxide synthase is a useful marker for complete restoration of the dystrophin associated complex and should be used as one of the criteria for selecting the recombinant molecule to be used for gene therapy in Duchenne muscular dystrophy.
杜氏肌营养不良症是一种致命的X连锁肌病，由于缺乏有效的治疗方法，人们将注意力集中在了基因治疗的可能性上。基因治疗方法的目的是恢复肌营养不良蛋白相关蛋白复合物，其中一个成员是神经元型一氧化氮合酶，它是信号转导中的一种重要酶。利用转基因mdx小鼠以及人和鼠重组肌营养不良蛋白的质粒基因转移，来评估在不同表达水平的肌营养不良蛋白基因转移后，神经元型一氧化氮合酶是否能够恢复到肌膜上。使用小鼠回复纤维和患有不同肌营养不良蛋白缺失的人类患者，来评估外显子缺失与神经元型一氧化氮合酶在肌膜上定位缺失之间的关系。我们证明，外显子45 - 48编码的结构域是神经元型一氧化氮合酶定位于肌膜所必需的。基于这些观察结果，我们认为神经元型一氧化氮合酶是肌营养不良蛋白相关复合物完全恢复的一个有用标志物，应用作选择用于杜氏肌营养不良症基因治疗的重组分子的标准之一。
