A novel approach to identify Duchenne muscular dystrophy patients for aminoglycoside antibiotics therapy
A novel approach to identify Duchenne muscular dystrophy patients for aminoglycoside antibiotics therapy
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DOI：
10.1016/j.braindev.2004.09.014
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发表时间：
2005-09-01
影响因子：
1.7
通讯作者：
Miike, T
中科院分区：
文献类型：
--
作者：
Kimura, S;Ito, K;Miike, T
氨基糖苷类抗生素已被发现可抑制mdx小鼠中有缺陷的抗肌萎缩基因的无义突变，提示可能治疗杜氏肌营养不良症（DMD）。然而，寻找适合这种治疗的患者非常困难，因为：(1)只有5-13%的DMD患者存在肌营养不良蛋白基因无义突变；(2)由于肌营养不良蛋白cDNA非常长（14 kb），寻找该基因无义突变具有挑战性；(3)氨基糖苷诱导的读透效率取决于无义突变的种类。为了建立一个系统来识别有资格接受氨基糖苷治疗的候选人，我们从9名患有肌营养不良蛋白基因无义突变的DMD患者中分离出成纤维细胞，用AdMyoD诱导其分化为肌原性谱系，并暴露于大霉素中。用体外肌营养不良蛋白染色法和western blotting法检测庆大霉素暴露的肌管中肌营养不良蛋白的表达。结果表明，庆大霉素能够通过解读基因中无义突变TGA诱导分化肌管中肌营养不良蛋白的表达；没有发生无义突变TAA和TAG的通读，因此没有导致肌营养不良蛋白的表达。因此，推测氨基糖苷治疗对无义突变TGA的DMD患者的疗效远高于无义突变TAA和TAG的DMD患者。在这项研究中，我们引入了一个简单的系统来识别这种治疗的患者，并首次报道了在使用庆大霉素的DMD患者的肌管中检测到肌营养不良蛋白的表达。(c) 2004 Elsevier B.V.版权所有
Aminoglycoside antibiotics have been found to suppress nonsense mutations located in the defective dystophin gene in mdx mice, suggesting a possible treatment for Duchenne muscular dystrophy (DMD). However, it is very difficult to find patients that are applicable for this therapy, because: (1) only 5-13% of DMD patients have nonsense mutations in the dystrophin gene, (2) it is challenging to find nonsense mutations in the gene because dystrophin cDNA is very long (14 kb), and (3) the efficiency of aminoglycoside-induced read-through is dependent on the kind of nonsense mutation. In order to develop a system for identifying candidates that qualify for aminoglycoside therapy, fibroblasts from nine DMD patients with nonsense mutation of dystrophin gene were isolated, induced to differentiate to myogenic lineage by AdMyoD, and exposed with gentamicin. The dystrophin expression in gentamicin-exposed myotubes was monitored by in vitro dystrophin staining and western blotting analysis. The results showed that gentamicin was able to induce dystrophin expression in the differentiated myotubes by the read-through of the nonsense mutation TGA in the gene; a read-through of the nonsense mutations TAA and TAG did not occur and consequently did not lead to dystrophin expression. Therefore, it is speculated that the aminoglycoside treatment is far more effective for DMD patients that have nonsense mutation TGA than for patients that have nonsense mutation TAA and TAG. In this study, we introduce an easy system to identify patients for this therapy and report for the first time, that dystrophin expression was detected in myotubes of DMD patients using gentamicin. (c) 2004 Elsevier B.V. All rights reserved.