Adeno-associated virus-delivered artificial microRNA extends survival and delays paralysis in an amyotrophic lateral sclerosis mouse model.
Adeno-associated virus-delivered artificial microRNA extends survival and delays paralysis in an amyotrophic lateral sclerosis mouse model.
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DOI：
10.1002/ana.24618
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发表时间：
2016-04
影响因子：
11.2
通讯作者：
Sena-Esteves M
中科院分区：
文献类型：
--
作者：
Stoica L;Todeasa SH;Cabrera GT;Salameh JS;ElMallah MK;Mueller C;Brown RH Jr;Sena-Esteves M
肌萎缩侧索硬化症（ALS）是一种致命的神经退行性疾病，其特征是运动神经元的丧失，导致在诊断后五年内进行性肌无力、瘫痪和死亡。大约10%的病例是遗传性的，其中20%是由于超氧化物歧化酶1（SOD1）基因的突变。阿舒唑，唯一一种FDA批准的ALS药物，只能延长几个月的生存期。在转基因ALS小鼠模型中的实验显示，降低突变SOD 1蛋白的水平是一种潜在的治疗方法。我们试图开发一种有效的AAV介导的RNAi基因治疗ALS。将编码针对人SOD1的人工microRNA的单链AAV9载体注射到新生SOD1G93A小鼠的侧脑室中，并评估对疾病进展和存活的影响。这种疗法延长了50%的中位生存期，并延迟了后肢瘫痪，动物保持行走直到人道终点，这是由于体重迅速减轻。AAV9处理的SOD1G93A小鼠显示出上运动神经元和下运动神经元中突变体人SOD1 mRNA水平的降低，以及多个参数的显著改善，包括脊髓运动神经元的数量、前根轴突的直径和SOD1G93A脊髓中神经炎症的程度。小鼠还显示出先前未探索的肺功能变化，AAV9处理的SOD1G93A小鼠显示出令人联想到患者病理生理学的表型。这些研究清楚地表明，AAV9递送的SOD1特异性人工microRNA是ALS的有效且可转化的治疗方法。
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by loss of motor neurons, resulting in progressive muscle weakness, paralysis and death within five years of diagnosis. About 10% of cases are inherited, of which 20% are due to mutations in the superoxide dismutase 1 (SOD1) gene. Riluzole, the only FDA approved ALS drug, prolongs survival by only a few months. Experiments in transgenic ALS mouse models have shown decreasing levels of mutant SOD1 protein as a potential therapeutic approach. We sought to develop an efficient AAV mediated RNAi gene therapy for ALS. A single stranded AAV9 vector encoding an artificial microRNA against human SOD1 was injected into the cerebral lateral ventricles of neonatal SOD1G93A mice and impact on disease progression and survival assessed. This therapy extended median survival by 50% and delayed hindlimb paralysis, with animals remaining ambulatory until the humane endpoint, which was due to rapid body weight loss. AAV9-treated SOD1G93A mice showed reduction of mutant human SOD1 mRNA levels in upper and lower motor neurons and significant improvements in multiple parameters including the numbers of spinal motor neurons, diameter of ventral root axons, and extent of neuroinflammation in the SOD1G93A spinal cord. Mice also showed previously unexplored changes in pulmonary function, with AAV9-treated SOD1G93A mice displaying a phenotype reminiscent of patient pathophysiology. These studies clearly demonstrate that an AAV9-delivered SOD1-specific artificial microRNA is an effective and translatable therapeutic approach for ALS.