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

• 批准号: 8716885
• 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
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-16 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8716885
• 关键词: Acetylation; Acute; Adverse reactions; Affect; Amino Acid Transporter; Biological Assay; Blood - brain barrier anatomy; Brain; Brain-Derived Neurotrophic Factor; Chemosensitization; Complex; Copper; Corpus striatum structure; Coupled; DNA; DNA Methylation; DNA Methyltransferase; 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; Industrial fungicide; Ions; Iron; K-Series Research Career Programs; Knowledge; Long-Term Effects; Maneb; Mass Spectrum Analysis; 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; Plasma; 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; design; divalent metal; dopaminergic neuron; environmental agent; epigenome; epigenomics; histone modification; in vivo; in vivo Model; interest; meetings; methylome; neurotoxic; neurotoxicity; next generation; oxidation; pesticide exposure; programs; promoter; response; uptake

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 反复接触环境药物（例如农药和金属）可以改变基因表达 模式，可能导致对未来有毒损害的敏感性增强。超过90％ 帕金森氏病（PD）病例被认为是特发性，流行病学表明参与 PD中的农药。具体而言，暴露于除草剂帕拉奎特（PQ）和杀菌剂maneb（MB）是 与PD风险增加75-80％有关。另外，基因表达的表观基因组变化是 假设是在PD等神经退行性疾病中发挥作用。我以前的NIEHS资助研究（F32 ES019821）证明，PQ神经毒性的MB-抑制不仅是由于增强的氧化作用。 压力，但一种复杂的机制，涉及通过MB进行硫醇调整和PQ氧化。附加基因 表达数据表明，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个金属转运蛋白。特别感兴趣，ala-，ser-，cys- 偏爱转运蛋白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诱导的大于2倍。为此的工作假设 K99/R00职业发展奖提案是长期暴露于MB增强PQ介导的 由于修饰氨基酸引起的PQ摄取增加引起的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。为此，4 将使用由体外和体内模型组成的具体目标（2个指导，2个独立）。具体 AIM 1（K99指导阶段），将使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诱导引起的。特定目标2（K99指导阶段） 将研究与多巴胺代谢相关的基因的甲基化是否与MB- 增强的PQ神经毒性。在特定目标3（R00独立阶段）中提出的研究将探索 由长期农药暴露引起的组蛋白H3甲基化和乙酰化的改变。最后，具体 AIM 4（R00独立阶段）将研究DNA甲基的全局变化，以响应长期 术语农药暴露。该项目旨在进一步制定我的研究计划并实现战略 通过研究表观基因组学和 培训下一代环境科学家。拟议的研究将符合这些目标，并 促进我独立研究生涯的发展。