An in vivo multiplex model to study gene-environment interaction in Parkinson's Disease

项目摘要/摘要 帕金森病是一种以富含α突触核蛋白为特征的进行性神经退行性疾病 神经元内含物。最近的全基因组相关研究和流行病学研究已经 分别确定了多个候选基因和环境因素，它们可以改变帕金森病的风险。学习 由于缺乏模型系统，与环境因素的多基因相互作用一直是困难的。然而， 研究暗示，α-突触核蛋白、遗传风险因素和环境之间存在复杂的关系 各种因素。在我们的初步数据中，我们使用果蝇帕金森病模型建立了一个多维模型。在这 模型中，我们表达了人α-突触核蛋白，同时修饰了神经元中的候选基因，并揭示了 成虫飞向鱼藤酮。在该模型中使用可伸缩技术的组合，我们确定了新的交互 在α-突触核蛋白、环境因素和GWA3个基因中。最重要的假设是多元的。 模型结合IPSC来源的神经元可用于识别和研究IPSC的作用机制。 新的基因-环境相互作用。此外，该模型系统将识别潜在的药物靶点 可以改变基因与环境的相互作用。在目标1中，进行了一系列实验，包括超分辨率 显微镜和IPSC衍生的酪氨酸羟化酶(TH)神经元，将被用来表征 LRRK2、鱼藤酮和α-突触核蛋白之间的相互作用，这是用多重模型确定的。这些 实验将在主要导师梅尔·B·费尼的实验室进行。目标2将涉及理解 LRRK2、鱼藤酮和α-突触核蛋白的相互作用机制。以前的研究和初步研究 实验表明，肌动蛋白的超稳定性在调节神经毒性中起着核心作用。在此 将对果蝇和IPSC衍生的TH进行生化、免疫组织学和神经毒性分析 神经元(获得致病LRRK2-G2019S和保护性LRRK2-R1398H IPSCs)的作用研究 肌动蛋白在调节这种基因-环境相互作用中的动力学。这些实验将在Dr。 费尼的实验室。在独立的R00部分中，可以改变LRRK2之间相互作用的可药物靶标， 鱼藤酮和α-突触核蛋白将被鉴定。此外，我们将筛选其他与帕金森病相关的神经毒物 通过肌动蛋白超稳定作用与LRRK2和α-突触核蛋白相互作用。我们会从基因和药理上 抑制mRCKα，一种可以调节果蝇、ipsc衍生的神经元和小鼠肌动蛋白超稳定的激酶 模特。我的神经毒理学和神经变性训练将通过教学课程和 参加哈佛医学院临床病理学会议和Exposome新兵训练营，网址为 哥伦比亚大学由共同导师加里·米勒组织。该项目可能阐明一种新的模型系统 可用于鉴定和研究基因-环境相互作用的机制。我所接受的训练 将使我能够过渡到独立，并领导一个实验室研究 神经退行性疾病中的基因-环境相互作用。

Project Summary/Abstract Parkinson's disease (PD) is a progressive neurodegenerative disorder that is characterized by α-synuclein-rich neuronal inclusions. Recent genome-wide associated studies (GWAS) and epidemiological studies have identified multiple candidate genes and environmental factors, respectively, which can modify PD risk. Studying polygenic interactions with environmental factors has been difficult due to the lack of a model system. However, studies have hinted at a complex relationship between α-synuclein, the genetic risk factors, and environmental factors. In our preliminary data, we have established a multiplex model using the Drosophila model of PD. In this model, we express human α-synuclein, simultaneously modify GWAS candidate genes in neurons, and expose adult flies to rotenone. Using a combination of scalable techniques in this model, we identified novel interactions among α-synuclein, environmental factors, and GWAS genes. The overarching hypothesis is a multiplex model, in combination with iPSC-derived neurons, can be used to identify and study the mechanism of novel gene-environment interactions. Further, this model system will identify potential drug targets that can modify the gene-environment interactions. In Aim 1, a series of experiments, including super-resolution microscopy and iPSC-derived tyrosine hydroxylase (TH) neurons, will be performed to characterize the interaction among LRRK2, rotenone, and α-synuclein, which was identified using the multiplex model. These experiments will be performed in the laboratory of primary mentor Mel B. Feany. Aim 2 will involve understanding the mechanism of interactions among LRRK2, rotenone, and α-synuclein. Previous studies and preliminary experiments have shown that actin hyperstabilization plays a central role in regulating neurotoxicity. Herein biochemical, immunohistological, and neurotoxicity assays will be performed in Drosophila and iPSC-derived TH neurons (obtained disease-causing LRRK2-G2019S and protective LRRK2-R1398H iPSCs) to study the role of actin dynamics in regulating this gene-environment interaction. These experiments will be performed in Dr. Feany's lab. In the independent R00 section, a druggable target that can modify the interaction among LRRK2, rotenone, and α-synuclein will be identified. Further, we will screen for other PD-related neurotoxicants that interact with LRRK2 and α-synuclein through actin hyperstabilization. We will genetically and pharmacologically inhibit MRCKα, a kinase that can regulate actin hyperstabilization, in flies, iPSC-derived neurons, and a mouse model. My neurotoxicology and neurodegeneration training will be facilitated by didactic courses and participation in Clinical Pathological conferences at Harvard Medical School and the Exposome boot camp at Columbia University organized by co-mentor Gary Miller. This project may elucidate a novel model system that can be used to identify and study the mechanism of gene-environment interactions. The training that I undertake will enable me to transition to independence and lead a laboratory investigating the molecular mechanisms of gene-environment interactions in neurodegenerative disorders.