Investigating environmental and gene-environment contributors to Parkinson's disease risk by coupling quantitative environmental exposure data to iPSC modeling

项目摘要 帕金森氏病（PD）被认为是一种“人为”的大流行病，部分原因是工业化引发的 这导致生产需求增加，并增加了接触这种增长所产生的有毒物质的机会。尽管 随着重要遗传风险因素的发现，特发性PD的大量病因风险是 在环境方面，农药接触是一个明确和有记录的风险因素。因此，理解 遗传驱动因素和环境风险因素之间的相互作用有很大的潜力， PD的病因和治疗。 该项目将调查环境因素，特别是农药，如何影响 在对中脑多巴胺能（mDA）神经元的影响水平上，PD的发展是一种细胞类型，其损失是 PD病理学的标志这项工作是由一个范式，其中高度定量农药 来自加州的一个大型PD队列的暴露数据与使用患者干细胞的筛查平台相关联 （iPSC）衍生的mDA神经元。用该方法对53种农药进行了检测，其中10种农药与PD直接相关 对mDA神经元有毒。本论文的主要工作如下：（1）深入剖析top对mDA神经元的毒性机制， 重点关注氟乐灵暴露对线粒体和细胞骨架的影响;（2）探索 神经胶质细胞在调节毒性中的作用，使用三种方法询问其他PD相关农药如何改变星形胶质细胞 和小胶质细胞生物学来影响mDA神经元病理学;以及（3）使用群体水平的散发性神经元病理学模型。 PD在培养皿中建立一个平台，该平台将基于对以下各项的响应在功能上对来自约100个供体的细胞进行分层： 然后使用计算方法来重建哪些细胞系受到不成比例的影响。 这将在遗传上不同的样本中识别出共享的表型。 职业发展计划旨在支持候选人成为独立的目标 学术医学中心的研究者，在临床上治疗PD患者，同时领导研究 研究他们的疾病。在充满活力的研究环境中，强大的指导和协作团队 由支持神经学主席（Batchelor）领导，杰出的医生科学家， 从神经变性（Khurana，Selkoe）到公共卫生和流行病学（Ritz），再到翻译 神经科学（鲁宾，Studer，鲍威尔）。这个导师团队，沿着广泛的研究培训， 相关的课程，将定位成功的候选人。通过以下途径获得的机构资源 布里格姆妇女医院和哈佛大学将支持候选人的职业生涯， 这使得高影响力的贡献和合作努力可以实现。成功执行这一 一份提案将使候选人沿着一条独立的职业道路，作为一名医生科学家， 基因-环境相互作用在PD中的相互作用，以更好地治疗和预防这种疾病。

PROJECT SUMMARY Parkinson's Disease (PD) is considered a “man-made” pandemic, triggered in part by industrialization that has entailed increased production needs and exposure to toxicants resulting from that growth. Despite the discovery of important genetic risk factors, a substantial amount of etiologic risk for idiopathic PD is environmental, with pesticide exposure being a clear and documented risk factor. As a result, understanding the interaction between genetic drivers and environmental risk factors has significant potential to inform etiology and treatment of PD. This project will investigate how environmental factors, specifically pesticides, influence the development of PD at the level of effects on midbrain dopaminergic (mDA) neurons, a cell type whose loss is the hallmark of PD pathology. The work is informed by a paradigm in which highly quantitative pesticide exposure data from a large PD cohort in California is linked to a screening platform using patient stem cell (iPSC)-derived mDA neurons. Fifty-three pesticides were linked to PD with this method and ten proved directly toxic to mDA neurons. The proposed work will: (1) deeply dissect mechanisms of mDA neuron toxicity for top hits with a focus on mitochondrial and cytoskeletal consequences of trifluralin exposure; (2) explore the role of glia in modulating toxicity using a triculture approach to ask how other PD-associated pesticides alter astrocyte and microglial biology to influence mDA neuron pathology; and (3) use population-level modeling of sporadic PD in a dish to build a platform that will stratify cells from ~100 donors functionally based on responses to pesticides and then use computational methods to reconstruct which cell lines are disproportionately affected. This will identify shared phenotypes among genetically disparate samples. The career development plan is designed to support the candidate's goal of becoming an independent investigator at an academic medical center, treating PD patients in clinic while concurrently leading a research team studying their disease. A strong mentorship and collaborative team in a vibrant research environment is led by a supportive neurology chairperson (Batchelor), outstanding physician scientists with expertise spanning from neurodegeneration (Khurana, Selkoe) to public health and epidemiology (Ritz) to translational neuroscience (Rubin, Studer, Powell). This mentorship team, along with extensive research training and relevant coursework, will position the candidate for success. The institutional resources available through Brigham and Women's Hospital, and Harvard University will support the candidate's career in an environment that makes high impact contributions and collaborative endeavors achievable. The successful execution of this proposal will position the candidate along a path for an independent career as a physician scientist studying the interplay of gene-environment interactions in PD to better treat and prevent this disease.