Gentamicin administration in Duchenne patients with premature stop codon. Preliminary results.
Gentamicin administration in Duchenne patients with premature stop codon. Preliminary results.
复制标题
终止密码子过早的 Duchenne 患者使用庆大霉素。
作者：
L. Politano;G. Nigro;Nigro;G. Piluso;S. Papparella;O. Paciello;Comi Li
本研究的目的是研究庆大霉素给药是否可以恢复由提前终止密码子引起的杜氏肌营养不良症患者横纹肌中的肌营养不良蛋白表达，如mdx小鼠中所报告的。根据那不勒斯第二大学伦理委员会批准的方案，4名Duchenne患者，仍能走动或处于轮椅阶段不到4个月，从导致过早终止密码子的点突变患者中选择，接受两个6天周期的硫酸庆大霉素治疗，间隔7周。第二个周期给药后进行肌肉活检;通过免疫组织化学和蛋白质印迹法对标本进行分析。通过动态试验和肌酸激酶值监测骨骼肌变化;在治疗开始和结束时，通过心电图、超声心动图、声学密度测定和肺活量评价心脏和呼吸状态。还监测了肾毒性和耳毒性等副作用。四分之三的患者，谁拥有最宽容的UGA作为终止密码子，显示阳性结果。在一名患者中，有一个戏剧性的肌营养不良蛋白的重新表达的免疫组织化学和蛋白质印迹;在两名患者中，肌营养不良蛋白阳性纤维被视为由抗体的杆域与免疫组织化学;第四名患者，与UAA作为终止密码子，没有表现出肌营养不良蛋白的表达。这些结果表明，庆大霉素能够恢复无义突变的杜氏患者亚组中肌营养不良蛋白的表达，提高了肌营养不良症的第一种药物治疗的可能性。
Aim of the study was to investigate whether the administration of gentamicin could restore dystrophin expression in striated muscles of patients with Duchenne muscular dystrophy caused by premature stop codon, as reported in mdx mice. Four Duchenne patients, still ambulant or in wheelchair stage for less than 4 months, selected among those with point mutations resulting in premature stop codons, received two 6-day cycles of gentamicin sulfate, at an interval of 7 weeks, according to the protocol approved by the Ethics Committee of the Second University of Naples. A muscle biopsy was performed after the second cycle of administration; the specimens were analysed by both immuno-histochemistry and Western blotting. Skeletal muscle changes were monitored by dynamic tests and Creatine Kinase values; at the beginning and end of treatment, cardiac and respiratory status was evaluated by electrocardiography, echocardiography, acoustic densitometry and vital capacity. Side-effects such as nephrotoxicity and ototoxicity were also monitored. Three out of four patients, who had the most permissive UGA as stop codon, showed positive results. In one patient, there was a dramatic re-expression of dystrophin by both immuno-histochemistry and Western blot; in two patients, dystrophin positive fibres were seen by the antibody to the rod domain with immuno-histochemistry; the fourth patient, with UAA as stop codon, showed no expression of dystrophin at all. These results suggest that gentamicin is able to recover dystrophin expression in a subset of Duchenne patients with nonsense mutations, raising the possibility of the first pharmacological treatment for muscular dystrophy.