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

肌动蛋白结合蛋白 Coronin 1C 在帕金森病发病机制中的影响

• 批准号: 10392204
• 负责人: Birgitt Schuele
• 金额: $ 23.61万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-15 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10392204
• 关键词: Actin-Binding Protein; Address; Affect; Age; Bradykinesia; Candidate Disease Gene; Cell Line; Cell physiology; Clinical; Clinical Research; Clustered Regularly Interspaced Short Palindromic Repeats; Cytoskeleton; Disease; Endoplasmic Reticulum; Endosomes; Engineered Gene; Engineering; Etiology; Family; Gene Expression; Genes; Genetic; Goals; Guanosine Triphosphate Phosphohydrolases; Human; Human Engineering; Hyperactivity; In Vitro; Individual; Induced pluripotent stem cell derived neurons; Introns; LRRK2 gene; Lewy Body Disease; Lewy body pathology; Membrane; Mitochondria; Modeling; Modification; Molecular; Morphology; Mutation; Nerve Degeneration; Neurites; Neurodegenerative Disorders; Neurons; Onset of illness; Outcome; Parkinson Disease; Parkinsonian Disorders; Pathogenesis; Pathology; Pathway interactions; Pattern; Penetrance; Phenotype; Phosphorylation; Phosphotransferases; Point Mutation; Post-Translational Protein Processing; Proteins; Research; Risk; Risk Factors; Role; Signal Transduction; Single Nucleotide Polymorphism; Site; Symptoms; System; Testing; Tremor; Variant; alpha synuclein; base; coronin protein; disorder risk; gene repression; genetic risk factor; genetic testing; genetic variant; genome wide association study; genome-wide; in vitro Model; induced pluripotent stem cell; induced pluripotent stem cell technology; knock-down; loss of function; mitochondrial dysfunction; nerve stem cell; neuropathology; novel; overexpression; polygenic risk score; posture instability; rac1 GTP-Binding Protein; risk variant; tau Proteins; tau expression; therapeutic target; tool

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.

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PD的患病率估计为每10万人中有572人 45岁以上的人，估计到2030年美国将有120万例病例。虽然PD的确切原因不是 众所周知，许多因素，包括环境和遗传因素，已被确定并归因于 增加患PD的风险。在过去的二十年里，许多遗传形式的PD已经被描述在 除了通过候选基因研究和全基因组关联研究（GWAS）确定的风险基因外， 并且已经描述了超过90个独立的全基因组显著风险信号。这种遗传形式允许 用于识别机制和途径，并且当验证时，它们可以成为治疗靶点。 PD最常见的遗传形式是LRRK 2基因突变。LRRK 2是一种多结构域蛋白， GT3结构域和激酶酶结构域两者。虽然LRRK 2-PD被认为存在更多的 由于其单一遗传原因，其同质临床和神经病理学模式仍然存在显著差异， LRRK 2-PD之间的差异，这提出了几个关键问题：为什么不是所有的LRRK 2变异携带者 出现PD（降低的潜伏期）？为什么发病年龄甚至在家庭内也有很大差异 （发病年龄不同）？为什么LRRK 2-PD病例之间的神经病理学不同，即使在LRRK 2-PD病例中， 相同的等位基因变体（多形性病理学）？ 在这个项目中，我们的目标是开发一个系统来测试LRRK 2-PD背景下的遗传修饰剂，我们 命名了CORO 1C基因的一个变体，该变体最近在第一个LRRK 2 GWAS研究中发现。我们将 联合收割机先进的规则间隔短回文重复序列（CRISPR）和诱导 多能干细胞（iPSC）技术编辑单点突变和调节基因表达， CORO1C。这将使我们能够产生一个人类iPSC系的等基因组，其中我们编辑了一个点， 突变并激活或抑制CORO 1C表达以探测健康人中功能获得或丧失表型 和LRRK 2人iPSC神经元。我们的目标是：目标1：获得这样的人类CRISPR工程iPSC工具; 目的2：评估CORO 1C影响细胞功能和α-突触核蛋白或tau蛋白表达的程度， 目的3：确定CORO 1C对LRRK 2功能的影响程度。 这些结果将对定义与神经变性相关的新机制产生关键影响， 在功能上验证CORO 1C作为基于临床研究的基因组提名的PD新疾病靶点- 广泛的协会。这种从内源基因座上调/下调基因的体外方法结合了 与特定疾病相关的SNP基因编辑，并可以作为一个试验平台，以解开功能性 遗传风险因素作为PD和相关路易体病理学的修饰因子的后果。