Rescuing ⍺-synuclein toxicity through neuron-specific enhancement of autophagy

项目摘要 该提案概述了一个为期五年的职业发展计划，旨在促进主要研究者， 作为基础和转化神经科学的临床科学家，研究独立性，专攻细胞 类型特异性机制，调节神经元自噬和修改帕金森病（PD）的蛋白毒性。 申请人：申请人已完成医学博士学位。和博士学位，神经病学住院医师培训和奖学金 PD和运动障碍方面的培训。他以前有神经科学研究的经验， 小鼠模型研究脊髓延髓肌萎缩症中的自噬。他的职业发展计划旨在 专注于PD发病机制的研究，并在生物信息学，CRISPR基因编辑和 高通量筛选和先进的成像技术。在此过程中， 建立在他以前的训练，以实现研究的独立性。他将受益于不断的指导 参与、定期会议和持续合作。他将学习更多的研究技术，通过 正式课程、讲习班和全国会议。这些培训机制将为申请人提供 具有在神经退行性研究中独立职业所需的科学和概念技能。 研究计划：PD是一种以运动和非运动/多系统为特征的进行性神经退行性疾病 导致严重残疾的症状目前尚无有效的疾病缓解疗法。 PD中的神经变性与β-突触核蛋白的毒性聚集有关，越来越多的证据表明，β-突触核蛋白的毒性聚集与PD的神经变性有关。 突触核蛋白可以通过自噬的保守途径降解。然而，目前的调节方法 自噬不能在患者中赋予神经保护作用。在最近发表的工作中，申请人确定 MTMR 5（肌微管蛋白相关磷酸酶5，由SBF 1转录本编码）作为一种有效的神经元自噬 神经元中的抑制因子MTMR 5敲低增强神经元对诱导自噬的敏感性， 加速多种自噬底物的降解，包括疾病相关的和聚集的， 倾向性蛋白质与此相一致，该提案将测试减少神经元中的MTMR 5的中心假设， 增强β-突触核蛋白的自噬清除并减少β-突触核蛋白相关的神经元死亡。申请人 将使用人类诱导多能干细胞（iPSC）来确定操纵SBF 1/MTMR 5是否能增强细胞增殖。 通过自噬的突触核蛋白周转（Aim 1）和改变β-突触核蛋白的蛋白毒性（Aim 2）。他还将雇用 无偏见的，基于全基因组CRISPR的筛选，以揭示调节神经元中MTMR 5的关键因素（Aim 3）。 总之，这些研究建立了一个新的研究平台，专注于神经元自噬，肌管蛋白 生物学和PD及相关神经退行性疾病的治疗设计。

Project Summary This proposal outlines a five-year career development program aimed at promoting the principal investigator to research independence as a clinician scientist in basic and translational neuroscience, with specialization in cell type-specific mechanisms that regulate neuronal autophagy and modify proteotoxicity in Parkinson disease (PD). Applicant: The applicant has completed M.D. and Ph.D. degrees, residency training in neurology, and fellowship training in PD and movement disorders. He has previous experience in neuroscience research using cell and mouse models to study autophagy in spinobulbar muscular atrophy. His career development plan is designed to focus his research on PD pathogenesis and advance his knowledge in bioinformatics, CRISPR gene editing and high-throughput screening, and advanced imaging techniques. In so doing, the training plan outlined herein builds upon his prior training to achieve research independence. He will benefit from continual mentor engagement, regular meetings and ongoing collaborations. He will learn additional research techniques through formal coursework, workshops, and national meetings. These training mechanisms will provide the applicant with the scientific and conceptual skillset necessary for an independent career in neurodegenerative research. Research Plan: PD is a progressive neurodegenerative disorder marked by motor and non-motor/multi-systemic symptoms that lead to profound disability. There is no effective disease-modifying therapy currently available. Neurodegeneration in PD relates to toxic aggregation of ⍺-synuclein, and mounting evidence shows that ⍺- synuclein can be degraded through the conserved pathway of autophagy. However, current methods to modulate autophagy fail to confer neuroprotective effects in patients. In recently published work, the applicant identified MTMR5 (myotubularin-related phosphatase 5, encoded by the SBF1 transcript) as a potent neuronal autophagy suppressor in neurons. MTMR5 knockdown enhances the sensitivity of neurons to induction of autophagy, and accelerates the degradation of multiple autophagy substrates, including disease-associated and aggregate- prone proteins. In line with this, this proposal will test the central hypothesis that reducing MTMR5 in neurons augments autophagic clearance of ⍺-synuclein and reduces ⍺-synuclein-related neuronal death. The applicant will use human induced pluripotent stem cells (iPSCs) to determine if manipulating SBF1/MTMR5 enhances ⍺- synuclein turnover via autophagy (Aim 1) and modifies ⍺-synuclein proteotoxicity (Aim 2). He will also employ unbiased, genome-wide CRISPR-based screens to uncover key factors regulating MTMR5 in neurons (Aim 3). Collectively, these studies establish a novel research platform focusing on neuronal autophagy, myotubularin biology, and therapy design in PD and related neurodegenerative disorders.