Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive motor neuron (MN) death. Mutation of the superoxide dismutase 1 (SOD1) gene, which results in abnormal protein aggregation, is one of the causes of familial ALS. Autophagic dysfunction occurs in SOD1-G93A mutant mice as the disease progresses, but the etiology of this disease is still unclear. Optineurin (OPTN) is an adaptor that is involved in autophagy and participates in aggrephagy and mitophagy. Previous studies have established that OPTN mutations contribute to diseases such as glaucoma and ALS. However, the function of OPTN in autophagy and mitophagy has not been intensively investigated in models of ALS. In this study, we assessed the beneficial effect of OPTN on autophagy and mitochondrial function by intrathecally injecting adeno-associated virus 9 (AAV9)-OPTN into SOD1-G93A transgenic mice and by administering lentivirus (LV)-OPTN to cells expressing the SOD1-G93A mutant protein. The expression of voltage-dependent anion channel 1 (VDAC1) was increased and autophagy was elevated after OPTN gene therapy, as shown by a lower level of p62 and a higher level of microtubule-associated protein 1A/1B-light chain 3 (LC3)-II. Moreover, using electron microscopy, we observed a hyperpolarized mitochondrial transmembrane potential and reversal of mitochondrial morphological abnormalities. Furthermore, the protein level of TANK-binding kinase 1 (TBK1) was increased, suggesting that mitophagy was increased. Our findings from both animal and cell line studies strongly suggest that OPTN gene therapy is a powerful strategy to increase autophagy and protect mitochondria to prevent the progression of ALS and could be effective in the treatment of ALS.

中文翻译：

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OPTN 基因治疗可增加表达 SOD1-G93A 的转基因小鼠和细胞的自噬并保护线粒体

肌萎缩侧索硬化症 (ALS) 是一种神经退行性疾病，其特征是进行性运动神经元 (MN) 死亡。超氧化物歧化酶1（SOD1）基因突变导致蛋白质异常聚集，是家族性ALS的病因之一。随着疾病的进展， SOD1 -G93A突变小鼠会出现自噬功能障碍，但该疾病的病因仍不清楚。Optineurin (OPTN) 是一种参与自噬的接头，参与聚合自噬和线粒体自噬。先前的研究已证实OPTN突变会导致青光眼和 ALS 等疾病。然而， OPTN在自噬和线粒体自噬中的功能尚未在 ALS 模型中得到深入研究。在这项研究中，我们通过将腺相关病毒9（AAV9）-OPTN鞘内注射到SOD1 - G93A转基因小鼠中，并向表达SOD1-G93A的细胞施用慢病毒（LV）-OPTN，评估了OPTN对自噬和线粒体功能的有益影响。 G93A突变蛋白。OPTN基因治疗后，电压依赖性阴离子通道 1 (VDAC1) 的表达增加，自噬增强，p62 水平降低，微管相关蛋白 1A/1B-轻链 3 (LC3) 水平升高-II。此外，使用电子显微镜，我们观察到超极化的线粒体跨膜电位和线粒体形态异常的逆转。此外，TANK 结合激酶 1 (TBK1) 的蛋白水平增加，表明线粒体自噬增加。我们的动物和细胞系研究结果强烈表明，OPTN基因治疗是一种增强自噬和保护线粒体以防止 ALS 进展的强大策略，并且可以有效治疗 ALS。