Dieldrin-induced differential gene methylation and parkinsonian toxicity (R01ES031237)

狄氏剂诱导的差异基因甲基化和帕金森毒性 (R01ES031237)

• 批准号: 10709194
• 负责人: Alison Bernstein
• 金额: $ 36.03万
• 依托单位: RUTGERS BIOMEDICAL AND HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-11-10 至 2025-10-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10709194
• 关键词: 3-Dimensional; Adult; Affect; Biological; Biological Assay; Brain; Candidate Disease Gene; Cell Culture Techniques; Corpus striatum structure; DNA Modification Process; DNMT3a; Data; Development; Dieldrin; Disease; Disease susceptibility; Dopamine; Elderly; Environment; Environmental Risk Factor; Epidemiology; Epigenetic Process; Etiology; Exposure to; Functional disorder; Funding; Gene Expression; Gene Expression Profile; Genes; Genetic; Goals; HDAC9 gene; High Pressure Liquid Chromatography; Immunohistochemistry; In Situ; In Vitro; Individual; Inherited; Injections; Knowledge; Link; Luciferases; Mass Spectrum Analysis; Mediating; Mediator; Methods; Methylation; Mission; Modeling; Modification; Mus; Mutation; NR4A2 gene; National Institute of Environmental Health Sciences; Neurites; Neurons; Parkinson Disease; Parkinsonian Disorders; Pathogenesis; Pathologic; Pathway Analysis; Pathway interactions; Periodicity; Persons; Phenotype; Play; Predisposition; Presynaptic Terminals; Radioactive; Risk; Risk Factors; Role; Scanning; Sequencing By Hybridizations; Site-Directed Mutagenesis; Specificity; System; Testing; Toxic Environmental Substances; Toxic effect; Western Blotting; alpha synuclein; candidate identification; cell type; cytotoxicity; dopaminergic neuron; epidemiology study; epigenome; experimental study; genome-wide; genomic locus; in vivo; in vivo Model; laser capture microdissection; male; methylation pattern; motor behavior; mouse model; neurotransmission; nigrostriatal pathway; novel; offspring; organochlorine pesticide; pesticide exposure; pre-formed fibril; response; sex; sporadic Parkinson' s Disease; synaptic function; synucleinopathy; tool; toxicant; transcriptome sequencing; uptake

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.

项目概要 大多数帕金森病 (PD) 病例并非由遗传性单基因突变引起， 疾病病因学涉及遗传和环境因素的结合。流行病学研究显示 接触杀虫剂，特别是狄氏剂等有机氯杀虫剂，会增加散发性感染的风险 PD。在PD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 成人接触多巴胺能毒物 MPTP 的易感性增加，并且根据我们的新数据 NIEHS 资助的 R21，α-突触核蛋白 (α-syn) 预形成原纤维 (PFF)。表观基因组是这一过程的潜在调节者 发育暴露、神经元脆弱性增加和成人疾病之间的关系。排队 有了这个想法，我们最近确定了响应发育的性别特异性差异甲基化模式 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 导致基因表达和表型的变化持续到成年，改变敏感性 帕金森病的侮辱并促进帕金森病的发展。 为了验证这一假设，我们将确定细胞类型特异性 DNA 修饰和表达谱 先前在狄氏剂模型中确定了候选基因（目标 1）；分析突触末端的功能 我们新颖的狄氏剂/PFF 两次打击模型（目标 2）；并确定狄氏剂或候选基因的表达是否改变 影响多巴胺能神经元细胞培养模型中对 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发挥作用 在帕金森毒性的生物反应中的功能作用。 完成这些目标将进一步履行 NIEHS 的使命，即增加我们对环境如何影响的了解。 影响人们以促进更健康的生活，具体项目目标是将暴露与 基因表达、神经元表型和 PD 易感性的功能变化。提出的实验 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机制。这个项目 还将通过建立一个内部模型来扩展我们用于询问表观遗传变化功能的工具库。 将特定表观遗传机制与帕金森毒性联系起来的体外实验范例。与我们在 结合发育暴露和成人 PFF 注射的体内模型，我们将有一组实验 适当的系统将使我们能够测试各种暴露以及暴露的组合， 体内和体外。总之，这套工具将使我们能够探索 PD-的机制。 相关暴露改变了 PD 中的神经元脆弱性，进一步推进了 NIEHS 的目标，即了解如何结合 暴露影响疾病发病机制和个体易感性。