Investigating Mic60 as a novel gene therapy target for Parkinson's disease

摘要 目前，还没有治疗方法可以阻止或阻止帕金森病（PD）的无情进展， 迫切需要确定能够改变疾病进程的治疗方法。基因转移的应用 神经变性的治疗是一个新兴的研究领域，有可能直接解决 在细胞水平上进行性神经元损失。关键是要确定基因疗法，将恢复细胞 健康和功能，从而改变疾病进程。虽然PD的具体病因仍然难以捉摸， 有证据表明，早期线粒体呼吸和稳态功能障碍在PD中起主要作用， 发病机制这项建议的研究重点是评估必要的线粒体的潜力， 结构蛋白Mic60，也被称为丝裂素，作为开发新的、可翻译的神经保护剂的途径 PD的策略。 Mic60是一种更大的线粒体蛋白复合物的重要组成部分，对于维持线粒体的功能至关重要。 结构和功能，以及Mic60的缺失可以在精神上影响线粒体稳态。Mic60功能 通过与PD相关蛋白、线粒体PINK1和Parkin的相互作用来调节， 线粒体健康和周转。我们以前发现，在细胞内Mic60蛋白水平降低， PD多巴胺毒性模型。我们也有证据表明PD患者大脑中的Mic60丰度较低。 此外，我们发现Mic60过表达可保护PD相关毒物，包括线粒体 有毒的鱼藤酮我们推测，所观察到的Mic60过表达的保护作用通过以下途径发生： 稳定线粒体功能。 我们建议通过以下目的研究Mic60作为PD基因治疗候选者的潜力： 目的1，我们将确定病毒介导的Mic60过表达在体内的理想剂量和长期安全性， 输送到黑质。在目标2中，我们将在体内测试Mic60过表达的神经保护作用 在PD的毒性和遗传模型中，评估Mic60作为靶向神经保护基因的潜力 治疗PD。我们的最终目标是建立在我们的体外数据和评估Mic60开发一种新的， 神经保护性PD治疗。由于线粒体功能障碍是PD和其他疾病的关键因素， 神经退行性疾病，通过Mic60靶向线粒体完整性以改善神经元健康， 为遗传性和散发性PD以及可能的其他神经系统疾病提供治疗。

ABSTRACT At present, there are no treatments that hinder or halt the inexorable progression of Parkinson disease (PD), and there is a critical need to identify treatments that can alter the course of the disease. The use of gene-transfer therapy for neurodegeneration is a burgeoning field of research that has potential for directly addressing progressive neuron loss at the cellular level. The key is to identify genetic therapies that will restore cellular health and function, thereby altering the disease course. While the specific etiology of PD remains elusive, evidence suggests that early dysfunction of mitochondrial respiration and homeostasis play a major role in PD pathogenesis. The research focus of this proposal is to evaluate the potential of the essential mitochondrial structural protein Mic60, also known as mitofilin, as an avenue to develop novel, translatable neuroprotective strategies for PD. Mic60 is a crucial component of a larger mitochondrial protein complex vital for maintaining mitochondrial architecture and function, and loss of Mic60 can detrimentally affect mitochondrial homeostasis. Mic60 function is regulated by interactions with proteins associated with PD, mitochondrial PINK1 and Parkin, which police mitochondrial health and turnover. We previously found that cellular levels of Mic60 protein are decreased in the dopamine toxicity model of PD. We also have evidence that Mic60 abundance is lower in PD patient brain. Further, we found that Mic60 overexpression protects against PD-relevant toxicants, including the mitochondrial poison rotenone. We hypothesize that the observed protective effects of Mic60 overexpression occur via stabilization of mitochondrial function. We propose to investigate the potential of Mic60 as a gene therapy candidate for PD via the following aims: In Aim 1, we will determine the ideal dose and long-term safety of viral-mediated Mic60 overexpression in vivo for delivery to the substantia nigra. In Aim 2, we will test the neuroprotective effect of Mic60 overexpression in vivo in toxicity and genetic models of PD, to evaluate the potential of Mic60 as a targeted neuroprotective gene therapy for PD. Our ultimate goal is to build on our in vitro data and evaluate Mic60 for development of a novel, neuroprotective PD treatment. As mitochondrial dysfunction is a key contributor to PD and other neurodegenerative diseases, targeting mitochondrial integrity via Mic60 to improve neuronal health may yield a therapy for both genetic and sporadic forms of PD, and possibly other neurological disorders.