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

摘要 目前，还没有任何治疗方法可以阻止或阻止帕金森病(PD)的不可避免的发展，以及 迫切需要确定能够改变疾病进程的治疗方法。基因转移技术的应用 神经退行性变的治疗是一个新兴的研究领域，有可能直接解决 细胞水平的进行性神经元丢失。关键是找出基因疗法，使细胞 健康和功能，从而改变疾病的进程。虽然帕金森病的具体病因仍然难以捉摸， 有证据表明，线粒体呼吸和动态平衡的早期功能障碍在帕金森病中起主要作用 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mitofin，作为开发新的、可翻译的神经保护的途径 帕金森病的策略。 Mic60是一个更大的线粒体蛋白质复合体的重要组成部分，对维持线粒体至关重要 结构和功能，以及Mic60的丢失都会对线粒体的动态平衡产生不利影响。Mic60函数 是由与PD、线粒体PINK1和Parkin相关的蛋白质相互作用调节的 线粒体健康和周转。我们之前发现，细胞中Mic60蛋白的水平在 帕金森病的多巴胺毒性模型。我们也有证据表明，帕金森病患者大脑中Mic60的丰度较低。 此外，我们发现Mic60的过度表达对PD相关毒物具有保护作用，包括线粒体 毒鱼藤酮。我们假设，观察到的Mic60过度表达的保护作用是通过 线粒体功能的稳定。 我们建议通过以下目标来研究Mic60作为帕金森病基因治疗候选基因的潜力： 目的1、我们将确定病毒介导的Mic60体内过表达的理想剂量和长期安全性 递送到黑质。在目标2中，我们将在体内测试Mic60过表达的神经保护作用 在帕金森病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 帕金森病的治疗。我们的最终目标是建立在我们的体外数据基础上，并评估Mic60用于开发一种新型的， 神经保护性帕金森病治疗。因为线粒体功能障碍是帕金森病和其他疾病的关键因素 通过Mic60靶向线粒体完整性以改善神经元健康的神经退行性疾病可能 产生一种治疗遗传性和散发性帕金森病的方法，可能还有其他神经疾病。

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.