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

项目总结/摘要 帕金森病（Parkinson's disease，PD）是一种进行性神经退行性疾病，以α-突触核蛋白（α-synuclein，α-synuclein）异常为特征。 神经内含物最近的全基因组相关研究（GWAS）和流行病学研究 分别确定了多个候选基因和环境因素，可以改变PD风险。研究 由于缺乏模型系统，多基因与环境因子的相互作用一直是困难的。然而，在这方面， 研究表明，α-突触核蛋白、遗传风险因素和环境因素之间存在复杂的关系。 因素在我们的初步数据，我们已经建立了一个多重模型，使用果蝇模型的PD。在这 模型中，我们表达人α-突触核蛋白，同时修饰神经元中的GWAS候选基因，并暴露 成虫对鱼藤酮过敏。在这个模型中使用可扩展技术的组合，我们确定了新的相互作用 α-突触核蛋白、环境因素和GWAS基因之间的关系。最重要的假设是 模型，与iPSC衍生的神经元相结合，可用于识别和研究 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，一系列实验，包括超分辨率 将进行显微镜和iPSC衍生的酪氨酸羟化酶（TH）神经元来表征 LRRK2、鱼藤酮和α-突触核蛋白之间的相互作用，这是使用多重模型鉴定的。这些 实验将在主要导师Mel B的实验室进行。Feany.目标2将涉及理解 LRRK2、鱼藤酮和α-突触核蛋白之间的相互作用机制。以往的研究和初步 实验已经表明肌动蛋白超稳定在调节神经毒性中起中心作用。本文 将在果蝇和iPSC衍生的TH中进行生物化学、免疫组织学和神经毒性测定。 神经元（获得的致病LRRK2-G2019S和保护性LRRK2-R1398H iPSC）来研究 肌动蛋白动力学调节这种基因-环境相互作用。这些实验将在Dr. 费尼的实验室在独立的R00部分，一个可以改变LRRK2， 鱼藤酮和α-突触核蛋白。此外，我们将筛选其他与PD相关的神经毒物， 通过肌动蛋白超稳定化与LRRK2和α-突触核蛋白相互作用。我们将从基因上 在果蝇、iPSC衍生的神经元和小鼠中抑制MRCK α（一种可调节肌动蛋白超稳定的激酶） 模型我的神经毒理学和神经变性培训将通过教学课程来促进， 参加哈佛医学院的临床病理学会议和 哥伦比亚大学组织的共同导师加里米勒。该项目可能阐明一种新的模型系统， 可用于识别和研究基因-环境相互作用的机制。我接受的训练 这将使我能够过渡到独立，并领导一个实验室研究分子机制， 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.