Altered transport and epigenomic changes in maneb-potentiated neurotoxicity

代森锰增强神经毒性的转运和表观基因组变化

基本信息

批准号：
8913968
负责人：
James R Roede
金额：
$ 24.9万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-16 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8913968
关键词：
Acetylation Acute Adverse reactions Affect Amino Acid Transporter Blood - brain barrier anatomy Brain Brain-Derived Neurotrophic Factor Chemosensitization Complex Copper Corpus striatum structure Coupled DNA DNA Methylation DNA Modification Methylases DNA-Binding Proteins DNMT3B gene DNMT3a Data Deposition Development Disease Dopamine Drug effect disorder Environmental Exposure Enzyme-Linked Immunosorbent Assay Epidemiology Exposure to Fostering Funding Future Gene Expression Gene Expression Profile Gene Expression Regulation Genes Goals Grant Herbicides High Pressure Liquid Chromatography Histone Acetylation Histone H3 In Vitro Inductively Coupled Plasma Mass Spectrometry Industrial fungicide Ions Iron K-Series Research Career Programs Knowledge Long-Term Effects Maneb Measures Mediating Mentors Metabolism Metals Methods Methylation Modification National Institute of Environmental Health Sciences Nerve Degeneration Neurodegenerative Disorders Neurons Neutral Amino Acid Transport Systems Oxidative Stress Paraquat Parkinson Disease Pesticides Phase Plasmids Play Population Predisposition Preparation Principal Investigator Probability Protons RNA Sequences Research Risk Role Scientist Small Interfering RNA Substantia nigra structure Sulfhydryl Compounds Testing Toxic effect Training Work abstracting aldehyde dehydrogenase 1 base career chromatin immunoprecipitation design divalent metal dopaminergic neuron environmental agent epigenome epigenomics histone methylation histone modification in vivo in vivo Model interest meetings methylome neurotoxic neurotoxicity next generation oxidation pesticide exposure programs promoter response transcriptome sequencing uptake

项目摘要

Abstract Repeated exposure to environmental agents, such as pesticides and metals, can alter gene expression patterns, potentially resulting in enhanced susceptibility to future toxic insults. With greater than 90% of Parkinson's disease (PD) cases considered idiopathic, epidemiology has indicated the involvement of pesticides in PD. Specifically, exposures to the herbicide paraquat (PQ) and the fungicide maneb (MB) are associated with a 75-80% increased risk of PD. Additionally, epigenomic changes in gene expression are postulated to play a role in neurodegenerative disorders like PD. My previous NIEHS funded research (F32 ES019821) demonstrated that MB-potentiation of PQ neurotoxicity was not simply due to enhanced oxidative stress, but a complex mechanism involving thiol-modification by MB and oxidation by PQ. Additional gene expression data show that MB significantly altered the expression of more genes compared to PQ. MB induced 4 different amino acid transporters and 2 metal transporters. Of particular interest, Ala-, Ser-, Cys- preferring transporter-1 (ASCT-1 (Slc1a4)) and proton coupled, divalent metal ion transporter (LSH (Slc11a1)) were induced greater than 2-fold by MB and PQ+MB compared to control. The working hypothesis for this K99/R00 Career Development Award proposal is that long-term exposure to MB potentiates PQ-mediated neurodegeneration of dopaminergic neurons caused by increased uptake of PQ due to modified amino acid transporter expression, metal-mediated oxidative stress and changes in the epigenome. To accomplish this, 4 specific aims (2 mentored, 2 independent) consisting of in vitro and in vivo models will be used. In Specific Aim 1 (K99 mentored phase), both in vitro and in vivo models will be employed to determine if potentiation of PQ neurotoxicity is due to MB-mediated induction of ASCT-1 and LSH. Specific Aim 2 (K99 mentored phase) will study if methylation of genes associated with dopamine metabolism is mechanistically involved in MB- potentiated PQ neurotoxicity. Research proposed in Specific Aim 3 (R00 independent phase) will explore alterations in histone H3 methylation and acetylation caused by long term pesticide exposure. Lastly, Specific Aim 4 (R00 independent phase) will investigate global alterations in the DNA methylome in response to long term pesticide exposure. This project is designed to further develop my research program and fulfill strategic goals of the NIEHS by connecting environmental influences to disease through the study of epigenomics and to train the next generation of environmental scientists. The proposed research will meet these goals and foster the development of my independent research career.

抽象的反复接触杀虫剂和金属等环境因素可能会改变基因表达模式，可能导致对未来有毒侮辱的敏感性增加。超过90%的帕金森病 (PD) 病例被认为是特发性的，流行病学表明涉及 PD 中的农药。具体而言，除草剂百草枯 (PQ) 和杀菌剂代森锰 (MB) 的暴露量为与 PD 风险增加 75-80% 相关。此外，基因表达的表观基因组变化是推测在 PD 等神经退行性疾病中发挥作用。我之前的 NIEHS 资助的研究（F32 ES019821）证明 MB 增强 PQ 神经毒性不仅仅是由于氧化增强应激，但涉及 MB 硫醇修饰和 PQ 氧化的复杂机制。附加基因表达数据显示，与 PQ 相比，MB 显着改变了更多基因的表达。 MB 诱导 4 种不同的氨基酸转运蛋白和 2 种金属转运蛋白。特别感兴趣的是 Ala-、Ser-、Cys- 优先选择转运蛋白 1 (ASCT-1 (Slc1a4)) 和质子耦合二价金属离子转运蛋白 (LSH (Slc11a1)) 与对照相比，MB 和 PQ+MB 诱导的诱导强度超过 2 倍。对此的工作假设 K99/R00 职业发展奖提案认为，长期接触 MB 会增强 PQ 介导的作用由于氨基酸修饰导致 PQ 摄取增加而引起多巴胺能神经元的神经变性转运蛋白表达、金属介导的氧化应激和表观基因组的变化。为了实现这一目标，4 将使用由体外和体内模型组成的具体目标（2 个指导的，2 个独立的）。具体来说目标 1（K99 指导阶段），将采用体外和体内模型来确定是否增强 PQ 神经毒性是由于 MB 介导的 ASCT-1 和 LSH 诱导所致。具体目标 2（K99 指导阶段）将研究与多巴胺代谢相关的基因甲基化是否在机械上参与 MB- 增强的 PQ 神经毒性。具体目标 3（R00 独立阶段）中提出的研究将探索长期接触农药引起的组蛋白 H3 甲基化和乙酰化的改变。最后，具体目标 4（R00 独立阶段）将研究 DNA 甲基化组的整体变化，以响应长期术语农药暴露。该项目旨在进一步发展我的研究计划并实现战略目标通过表观基因组学研究将环境影响与疾病联系起来，以实现 NIEHS 的目标培养下一代环境科学家。拟议的研究将实现这些目标促进我的独立研究事业的发展。