Dieldrin-induced differential gene methylation and parkinsonian toxicity

项目摘要 大多数帕金森病(PD)病例不是由遗传性单基因突变引起的，而且 疾病病因学涉及遗传和环境因素的组合。流行病学研究表明 接触杀虫剂，特别是接触狄氏剂等有机氯杀虫剂，会增加零星发病的风险。 警察。在帕金森病易感性增加的模型中，在发育过程中暴露于狄氏剂的小鼠表现出雄性特异性 成人暴露于多巴胺能毒物MPTP的易感性增加，在我们的新数据中 NIEHS资助的R21，α-突触核蛋白(α-SYN)预制纤维(PFF)。表观基因组是这一过程的潜在中介。 发育暴露、神经元易损性增加与成人疾病的关系。排队 有了这个想法，我们最近确定了性别特有的差异甲基化模式，以响应发育 狄氏剂暴露。我们假设狄氏剂在发育过程中诱导表观遗传修饰 导致持续到成年的基因表达和表型的变化，改变敏感度 对帕金森病患者的侮辱和对帕金森病发展的贡献。 为了验证这一假设，我们将确定细胞类型特定的DNA修饰和表达谱 先前在狄氏菌模型(AIM 1)中发现的候选基因；分析突触终末的功能 我们新的Dieldrin/PFF两次点击模型(Aim 2)；并确定Dieldrin或改变候选基因的表达 在多巴胺能神经元细胞培养模型中影响对α-syn pff的敏感性(目标3)。长期目标 这些实验的目的是确定狄氏剂相关的差异甲基化基因是否在 帕金森毒性的生物学反应中的功能作用。 这些目标的实现将进一步推动NIEHS的使命，即增加我们对环境如何 影响人们，以促进更健康的生活，并有一个具体的项目目标，将暴露于 基因表达、神经元表型和帕金森病易感性的功能变化。建议进行的实验 这将有助于建立一种将发育暴露与晚年疾病联系起来的生物学机制。这个项目 还将扩展我们的工具库，通过建立一个 将特定的表观遗传机制与帕金森病毒性联系起来的体外实验范式。随着我们的进入 结合发育暴露和成人PFF注射的活体模型，我们将有一套实验性的 建立系统，使我们能够测试各种曝光，以及曝光的组合， 无论是在体内还是体外。综上所述，这套工具将使我们能够探索PD- 相关暴露改变了帕金森病神经元的脆弱性，推动了NIEHS了解两者如何结合的目标 暴露会影响疾病的发病机制和个体的易感性。

Project Summary The majority of Parkinson’s disease (PD) cases are not caused by an inherited monogenic mutation and disease etiology involves a combination of genetic and environmental factors. Epidemiological studies show that pesticide exposure, particularly to organochlorine pesticides such as dieldrin, increases risk of sporadic PD. In a model of increased PD susceptibility, mice exposed to dieldrin during development show male-specific increased susceptibility to adult exposure to the dopaminergic toxicant MPTP and, in new data from our NIEHS-funded R21, α-synuclein (α-syn) preformed fibrils (PFFs). The epigenome is a potential mediator of this relationship between developmental exposures, increased neuronal vulnerability, and adult disease. In line with this idea, we recently identified sex-specific differential methylation patterns in response to developmental dieldrin exposure. We hypothesize that dieldrin-induced epigenetic modifications during development cause changes in gene expression and phenotype that persist into adulthood, altering the sensitivity to parkinsonian insults and contributing to the development of PD. To test this hypothesis, we will determine cell-type specific DNA modifications and expression profiles of previously identified candidate genes in the dieldrin model (Aim 1); analyze the function of synaptic terminals in our novel dieldrin/PFF two-hit model (Aim 2); and determine if dieldrin or altered expression of candidate genes affects susceptibility to α-syn PFFs in a dopaminergic neuron cell culture model (Aim 3). The long-term goal of these experiments is to determine whether dieldrin-associated differentially methylated genes play a functional role in the biological response to parkinsonian toxicity. Completion of these aims will further the mission of NIEHS to increase our understanding of how the environment affects people in order to promote healthier lives, with a specific project goal of connecting exposures with functional changes in gene expression, neuronal phenotype, and PD susceptibility. The experiments proposed here will help to establish a biological mechanism linking developmental exposure to late life disease. This project will also expand our repertoire of tools for interrogating the function of epigenetic changes by establishing an in vitro experimental paradigm to connect specific epigenetic mechanisms with parkinsonian toxicity. With our in vivo model that combines developmental exposure with adult PFF injections, we will have a set of experimental systems in place that will allow us to test a wide variety of exposures, as well as combinations of exposures, both in vivo and in vitro. Together, this suite of tools will enable us to explore the mechanisms by which PD- related exposures alter neuronal vulnerability in PD, furthering the goal of NIEHS to understand how combined exposures affect disease pathogenesis and individual susceptibility.