Functional amounts of dystrophin produced by skipping the mutated exon in the mdx dystrophic mouse
Functional amounts of dystrophin produced by skipping the mutated exon in the mdx dystrophic mouse
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DOI：
10.1038/nm897
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发表时间：
2003-08-01
期刊：
影响因子：
82.9
通讯作者：
Wilton, SD
中科院分区：
文献类型：
--
作者：
Lu, QL;Mann, CJ;Wilton, SD
作为基因治疗的靶标，杜氏肌营养不良症 (DMD) 存在许多障碍，但也为通过选择性剪接进行纠正提供了无与伦比的前景。抗肌营养不良蛋白基因的大部分突变发生在编码血影蛋白样中央杆状结构域的区域，该区域在很大程度上是可有可无的。因此，围绕突变的剪接可以产生缩短的但符合读框的转录本，从而允许部分功能性肌营养不良蛋白的翻译。我们通过将有效的转染方案与旨在促进突变外显子跳跃的 2-O-甲基化硫代磷酸反义寡核糖核苷酸 (2OMeAO) 相结合，在 mdx 营养不良小鼠（在肌营养不良蛋白基因的外显子 23 中携带突变）体内测试了这一想法。接受治疗的小鼠的大量肌纤维中持续产生正常水平的肌营养不良蛋白，并且显示出接受治疗的肌肉的功能改善。重复给药可增强肌营养不良蛋白的表达，但不会引发免疫反应。我们的数据证实了一种方法的现实实用性，原则上适用于大多数严重肌营养不良症病例。
As a target for gene therapy, Duchenne muscular dystrophy (DMD) presents many obstacles but also an unparalleled prospect for correction by alternative splicing. The majority of mutations in the dystrophin gene occur in the region encoding the spectrin-like central rod domain, which is largely dispensable. Thus, splicing around mutations can generate a shortened but in-frame transcript, permitting translation of a partially functional dystrophin protein. We have tested this idea in vivo in the mdx dystrophic mouse (carrying a mutation in exon 23 of the dystrophin gene) by combining a potent transfection protocol with a 2-O-methylated phosphorothioated antisense oligoribonucleotide (2OMeAO) designed to promote skipping of the mutated exon. The treated mice show persistent production of dystrophin at normal levels in large numbers of muscle fibers and show functional improvement of the treated muscle. Repeated administration enhances dystrophin expression without eliciting immune responses. Our data establishes the realistic practicality of an approach that is applicable, in principle, to a majority of cases of severe dystrophinopathy.