Molecular Mechanisms of Marine Organohalogen Bioaccumulation and Neurotoxicity

海洋有机卤素生物累积和神经毒性的分子机制

• 批准号: 10172906
• 负责人: AMRO M HAMDOUN
• 金额: $ 11.76万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10172906
• 关键词: ABCB1 gene; ABCG2 gene; Address; Benefits and Risks; Binding; Blood - brain barrier anatomy; Brain; CRISPR/Cas technology; Ca(2+)-Transporting ATPase; Cardiovascular system; Cells; Chemicals; Complex; Consumption; Crystallization; Data; Development; Ecosystem; Electrophysiology (science); Environment; Environmental Health; Etiology; Exposure to; Female; Fishes; Fluorescence; Future; Gender; Genes; Goals; Health; Heat Stress Disorders; Hippocampus (Brain); Human; Immune; Immune system; In Vitro; Infrastructure; Intestines; Kidney; Knowledge; Light; Link; Liver; Mediating; Mission; Modeling; Molecular; Molecular Target; Morphology; Multi-Drug Resistance; Mutation; Myopathy; National Institute of Environmental Health Sciences; Nervous system structure; Neurologic; Neurons; Neurotoxins; Oceans; Outcome; Pathway interactions; Penetration; Pharmaceutical Preparations; Physiological; Physiology; Polychlorinated Biphenyls; Population; Process; Protein Isoforms; Public Health; Relative Risks; Reticulum; Risk; Role; RyR1; Ryanodine; Ryanodine Receptors; Salmon; Sea Urchins; Seafood; Site; Structure; Structure-Activity Relationship; Testing; Time; Tissues; Toxic effect; Tuna; United States; Up-Regulation; Wild Type Mouse; Work; Xenobiotics; anthropogenesis; bioaccumulation; cancer cell; cancer type; cellular imaging; deep ocean; exposed human population; inhibitor/antagonist; male; man; muscular system; neurotoxicity; persistent organic pollutants; pollutant; pollutant interaction; receptor; risk minimization; screening; sex; uptake; voltage clamp

ABSTRACT There is urgent public health need to better understand the relative risks and benefits associated with consumption of seafood. The overall mission of this project is to understand the toxicity of marine organohalogen pollutants. We take a powerful approach to understanding and mitigating this risk by asking two questions, central to future efforts to predict and minimize risk. Aim 1 of this project asks how these compounds bioaccumulate, focusing on xenobiotic transporters, which are a key pathway for limiting accumulation of foreign chemicals. We will determine the interactions of the four major human xenobiotic transporters (XTs) with environmentally relevant natural and man-made marine organohalogens. The results will extend and expand the scope of our previous work indicating that several of these compounds can act as potent inhibitors of transporter function. In parallel, we will take advantage of recent progress with heterologous transporter-expression and CRISPR/CAS9 gene editing in sea urchins, to dissect the functional role of XTs in governing bioaccumulation in marine cells. This will be supported by a structure guided approach to determine how evolutionary changes in transporter structure modify interactions with TICs, following up on recent progress towards purification and crystallization of marine XTs in complex with pollutants. Aim 2 of this project will determine the structure activity relationships governing neurotoxicity of marine pollutants. These studies are motivated by preliminary data indicating that naturally produced organohalogens are highly potent inhibitors of ryanodine sensitive Ca2+ channels (RyRs) and Ca2+ ATPase transporters (SERCAs), which are arguably the most direct targets of environmentally relevant organohalogens in the brain. We will use primary cultures of hippocampal neurons cultured from male and female wild type mice to determine how activity at these molecular targets alter neuronal network Ca2+ dynamics and morphology using real-time fluorescence cell imaging and morphometric approaches. In addition, we will determine how hippocampal neurons that express mutation RyR1-R163C known to confer heat stress intolerance, alter sensitivity to organohalogens, and ask whether these effects are gender-specific. These studies will address the critical need to better understand the molecular mechanisms by which naturally occurring and man-made seafood pollutants accumulate in target cells and perturb the Ca2+ dynamics essential for normal neuronal network development.

摘要 公共卫生迫切需要更好地了解与以下方面相关的相对风险和益处： 海鲜的消费。本项目的总体使命是了解海洋有机卤素的毒性 污染物我们通过提出两个问题来理解和减轻这种风险， 未来的努力，以预测和尽量减少风险。该项目的目标1是研究这些化合物是如何生物积累的， 重点是异生物质转运蛋白，这是限制外来化学品积累的关键途径。我们 将确定四种主要的人类异生素转运蛋白（XTs）与环境的相互作用， 相关的天然和人造海洋有机卤素。这些结果将扩展和扩大我们的范围， 先前的工作表明这些化合物中的几种可以作为转运蛋白功能的有效抑制剂。在 同时，我们将利用异源转运蛋白表达和CRISPR/CAS9的最新进展， 海胆中的基因编辑，以剖析XTs在控制海洋细胞中生物积累方面的功能作用。 这将得到结构指导方法的支持，以确定转运蛋白的进化变化 结构修饰剂与TIC相互作用，跟踪最近在纯化和结晶方面的进展 海洋XTs与污染物的复合体。本项目的目标2将确定结构活性关系 控制海洋污染物的神经毒性。这些研究的动机是初步数据表明， 天然产生的有机卤素是兰尼碱敏感性Ca 2+通道（RyR）的高效抑制剂， Ca 2 + ATP酶转运蛋白（SERCA），可以说是环境相关的最直接的目标， 脑中的有机卤素我们将使用原代培养的海马神经元培养男性和女性 野生型小鼠，以确定这些分子靶标的活性如何改变神经元网络Ca 2+动力学， 使用实时荧光细胞成像和形态测量方法进行形态学研究。此外，我们将确定 海马神经元如何表达已知赋予热应激耐受性的突变RyR 1-R163 C， 对有机卤素的敏感性，并询问这些影响是否是性别特异性的。这些研究将解决 迫切需要更好地了解自然发生和人造海鲜的分子机制， 污染物在靶细胞中积累并扰乱正常神经元网络所必需的Ca 2+动力学 发展