Gentamicin-Induced Readthrough of Stop Codons in Duchenne Muscular Dystrophy
Gentamicin-Induced Readthrough of Stop Codons in Duchenne Muscular Dystrophy
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DOI：
10.1002/ana.22024
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发表时间：
2010-06-01
影响因子：
11.2
通讯作者：
Mendell, Jerry R.
中科院分区：
文献类型：
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作者：
Malik, Vinod;Rodino-Klapac, Louise R.;Mendell, Jerry R.
目的：本研究的目的是确定长期庆大霉素给药以实现终止密码子通读并产生全长肌营养不良蛋白的可行性。终止密码子的突变抑制，成功地实现了在mdx小鼠使用庆大霉素，代表了一个重要的发展中的治疗策略在杜氏肌营养不良症（DMD）。方法：两个DMD队列接受14天庆大霉素（7.5mg/kg/天）：队列1（n = 10）终止密码子患者和队列2（n = 8）移码对照。另外两个终止密码子DMD队列接受庆大霉素治疗（7.5 mg/kg）6个月：队列3（n = 12）每周给药一次，队列4（n = 4）每周给药两次。治疗前和治疗后的活组织检查进行了评估肌营养不良蛋白水平，作为临床outcomes.Results：在14天的研究，血清肌酸激酶（CK）下降了50%，这是没有看到移码DMD控制。庆大霉素治疗6个月后，肌营养不良蛋白水平显著升高（p = 0.027）;最高水平达到正常值的13%至15%（队列3中1例，队列4中2例），伴有血清CK降低，有利于药物诱导的终止密码子通读。这得到了力量稳定和用力肺活量略微增加的支持。治疗前稳定的转录本预测了庆大霉素治疗后肌营养不良蛋白的增加。通读效率不受终止密码子或其周围的第四个核苷酸的影响。在1例受试者中，抗原特异性干扰素-γ酶联免疫斑点试验检测到免疫原性抗肌萎缩蛋白epitope.Interpretation：结果支持努力实现药物诱导的突变抑制终止密码子。由通读产生的免疫原性表位强调了在抑制终止密码子的临床研究期间监测T细胞免疫的重要性。类似的原理也适用于其他分子策略，包括外显子跳跃和基因治疗。神经网络2010;67：771-780
Objective: The objective of this study was to establish the feasibility of long-term gentamicin dosing to achieve stop codon readthrough and produce full-length dystrophin. Mutation suppression of stop codons, successfully achieved in the mdx mouse using gentamicin, represents an important evolving treatment strategy in Duchenne muscular dystrophy (DMD).Methods: Two DMD cohorts received 14-day gentamicin (7.5mg/kg/day): Cohort 1 (n = 10) stop codon patients and Cohort 2 (n = 8) frameshift controls. Two additional stop codon DMD cohorts were gentamicin treated (7.5mg/kg) for 6 months: Cohort 3 (n = 12) dosed weekly and Cohort 4 (n = 4) dosed twice weekly. Pre- and post-treatment biopsies were assessed for dystrophin levels, as were clinical outcomes.Results: In the 14-day study, serum creatine kinase (CK) dropped by 50%, which was not seen in frameshift DMD controls. After 6 months of gentamicin, dystrophin levels significantly increased (p = 0.027); the highest levels reached 13 to 15% of normal (1 in Cohort 3, and 2 in Cohort 4), accompanied by reduced serum CK favoring drug-induced readthrough of stop codons. This was supported by stabilization of strength and a slight increase in forced vital capacity. Pretreatment stable transcripts predicted an increase of dystrophin after gentamicin. Readthrough efficiency was not affected by the stop codon or its surrounding fourth nucleotide. In 1 subject, antigen-specific interferon-gamma enzyme-linked immunospot assay detected an immunogenic dystrophin epitope.Interpretation: The results support efforts to achieve drug-induced mutation suppression of stop codons. The immunogenic epitope resulting from readthrough emphasizes the importance of monitoring T-cell immunity during clinical studies that suppress stop codons. Similar principles apply to other molecular strategies, including exon skipping and gene therapy. ANN NEUROL 2010;67:771-780