Impact of actin binding protein Coronin 1C in the pathogenesis of Parkinson's disease

帕金森病(PD)是一种进行性神经退行性疾病，通常以震颤为特征， 运动迟缓、僵硬和姿势不稳。据估计，每10万人中有572人患有帕金森病 45岁以上的人，估计到2030年美国将有120万病例。而帕金森病的确切原因并不是 已知，包括环境和遗传因素在内的多种因素已被确定并归因于 增加发生帕金森病的风险。在过去的二十年里，许多帕金森氏症的遗传形式被描述为 通过候选基因研究和全基因组关联研究(GWAS)确定的风险基因 已经描述了90多个独立的全基因组重大风险信号。这样的遗传形式使得 用于识别机制和途径，一旦得到验证，它们可能成为治疗的靶点。 帕金森病最常见的遗传形式是LRRK2基因的突变。LRRK2是一种多结构域蛋白，具有 GTP酶结构域和激酶酶结构域。而LRRK2-PD被认为呈现出更多的 同质性的临床和神经病理模式由于其单一的遗传原因，仍有惊人的表现 LRRK2-PD之间的差异，这提出了几个关键问题：为什么不是所有的LRRK2变异携带者 发展PD(降低外显率)？为什么发病年龄即使在家庭内部也会有很大的差异？ (发病时的可变年龄)？为什么LRRK2-PD病例之间的神经病理不同，即使在 相同的等位基因变异(多形性病理)？ 在这个项目中，我们的目标是开发一个系统，在LRRK2-PD的背景下测试遗传修饰物，我们 提名了CORO1C基因的一个变异，这是最近在第一个LRRK2 Gwas研究中发现的。我们会 结合高级成簇规则间隔短回文重复(CRISPR)和诱导 多能干细胞(IPSC)技术编辑单点突变并调节基因表达 CORO1C.这将允许我们生成人类IPSC线的等基因面板，在该面板中我们编辑了单个点 突变和激活或抑制CORO1C表达以检测健康人群功能表型的得失 和LRRK2人IPSC神经元。我们的目标是：目标1：衍生出这样的人类CRISPR工程IPSC工具； 目的2：评估CORO1C对细胞功能和α-突触核蛋白或tau表达的影响程度。 翻译后修饰；目标3：确定CORO1C对LRRK2功能的影响程度。 这些结果将对定义与神经退行性变相关的新机制和 从功能上验证CORO1C作为临床研究基因组提名的PD新的疾病靶点- 广泛的联想。这种结合内源基因上调/下调基因的体外方法 与特定疾病相关的SNP基因编辑，并可作为解开功能的试验床 遗传风险因素在帕金森病和相关的路易体病理中的修饰作用的后果。

Parkinson’s Disease (PD) is a progressive neurodegenerative disease and is commonly characterized by tremor, bradykinesia, stiffness, and postural instability. PD is estimated to be prevalent in 572 individuals per 100,000 individuals over 45, with an estimated 1.2 million cases in the US by 2030. While the exact cause of PD is not known, a multitude of factors, including environmental and genetic factors have been identified and attributed to increase the risk of developing PD. Many genetic forms of PD have been described in the last two decades in addition to risk genes identified through candidate gene studies and genome-wide association studies (GWAS) and over 90 independent genome-wide significant risk signals have been described. Such genetic forms allow for the identification of mechanisms and pathways, and they could become therapeutic targets when validated. The most common genetic forms of PD are mutations in the LRRK2 gene. LRRK2 is a multidomain protein with both a GTPase domain and a kinase enzymatic domain. While LRRK2-PD is thought to present with a more homogenous clinical and neuropathological pattern due to its single genetic cause, there are still striking differences among LRRK2-PD, which pose several critical questions: why is it that not all LRRK2 variant carriers develop PD (reduced penetrance)? Why can the age at disease onset be quite variable even within families (variable age at onset)? Why does the neuropathology vary between LRRK2-PD cases, even in cases with the same allelic variant (pleomorphic pathology)? In this project, we aim to develop a system to test genetic modifiers in the context of LRRK2-PD and we nominated a variant in the CORO1C gene, which was recently found in the first LRRK2 GWAS study. We will combine advanced Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and induced pluripotent stem cell (iPSC) technologies to edit a single point mutation and modulate gene expression of CORO1C. This will allow us to generate an isogenic panel of human iPSC lines, in which we edited a single point mutation and activate or inhibit CORO1C expression to probe for gain-or loss-of function phenotypes in healthy and LRRK2 human iPSC neurons. Our aims are: Aim 1: To derive such human CRISPR engineered iPSC tools; Aim 2: To assess to what extent CORO1C affects cellular function, and alpha-synuclein or tau expression and post translational modifications; Aim 3: To determine to what extent CORO1C has an impact on LRRK2 function. The outcomes will have a critical impact on defining new mechanisms related to neurodegeneration and functionally validate CORO1C as a new disease target for PD nominated by clinical research-based genome- wide association. This in vitro approach of up/down-regulation of genes from their endogenous loci combined with specific disease-associated SNP gene editing and could serve as a testbed to unravel the functional consequences of genetic risk factors as modifiers in PD and related Lewy body pathologies.