Mechanisms Controlling Sensitivity and Resistance to Dioxin-like Compounds: Role of AIP

控制对二恶英类化合物的敏感性和耐受性的机制：AIP 的作用

• 批准号: 10538943
• 负责人: Mark E Hahn
• 金额: $ 183.94万
• 依托单位: WOODS HOLE OCEANOGRAPHIC INSTITUTION
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-07 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10538943
• 关键词: ARA9 protein; Address; Affect; Agonist; Amino Acid Sequence; Aromatic Hydrocarbons; Aromatic Polycyclic Hydrocarbons; Aryl Hydrocarbon Receptor; Basic Science; Biochemical; COS-7 Cell; CRISPR/Cas technology; Cell Line; Cells; Chemical Exposure; Chemicals; DNA Resequencing; Dependence; Developmental Process; Dioxins; Disease Outcome; Dose; Embryo; Environmental Exposure; Environmental Pollution; Exhibits; Exposure to; Familial disease; Fishes; Fundulus heteroclitus; Gene Expression; Genes; Genetic; Genome; Goals; Hepatocyte; Hepatotoxicity; Homology Modeling; Human; In Vitro; Individual; Individual Differences; Indoles; Killifishes; Knock-in; Knowledge; Laboratories; Ligands; Measures; Mediating; Molecular; Molecular Chaperones; Mus; Mutation; National Institute of Environmental Health Sciences; Natural Products; Nuclear Translocation; Pathway interactions; Patients; Pituitary Gland Adenoma; Play; Polychlorinated Biphenyls; Population; Predisposition; Protein Deficiency; Proteins; Quantitative Trait Loci; Research; Resistance; Role; Signal Pathway; Specificity; Strategic Planning; Structure; System; Technology; Testing; Tetrachlorodibenzodioxin; Toxic effect; Variant; Zebrafish; aryl hydrocarbon receptor ligand; cellular engineering; design; developmental toxicity; epidemiology study; experience; experimental study; exposed human population; genome editing; genome-wide; human disease; in vitro Assay; in vivo; individual variation; insight; inter-individual variation; microbial; receptor function; resistance gene; response; toxicant; ARA9 protein; Address; Affect; Agonist; Amino Acid Sequence; Aromatic Hydrocarbons; Aromatic Polycyclic Hydrocarbons; Aryl Hydrocarbon Receptor; Basic Science; Biochemical; COS-7 Cell; CRISPR/Cas technology; Cell Line; Cells; Chemical Exposure; Chemicals; DNA Resequencing; Dependence; Developmental Process; Dioxins; Disease Outcome; Dose; Embryo; Environmental Exposure; Environmental Pollution; Exhibits; Exposure to; Familial disease; Fishes; Fundulus heteroclitus; Gene Expression; Genes; Genetic; Genome; Goals; Hepatocyte; Hepatotoxicity; Homology Modeling; Human; In Vitro; Individual; Individual Differences; Indoles; Killifishes; Knock-in; Knowledge; Laboratories; Ligands; Measures; Mediating; Molecular; Molecular Chaperones; Mus; Mutation; National Institute of Environmental Health Sciences; Natural Products; Nuclear Translocation; Pathway interactions; Patients; Pituitary Gland Adenoma; Play; Polychlorinated Biphenyls; Population; Predisposition; Protein Deficiency; Proteins; Quantitative Trait Loci; Research; Resistance; Role; Signal Pathway; Specificity; Strategic Planning; Structure; System; Technology; Testing; Tetrachlorodibenzodioxin; Toxic effect; Variant; Zebrafish; aryl hydrocarbon receptor ligand; cellular engineering; design; developmental toxicity; epidemiology study; experience; experimental study; exposed human population; genome editing; genome-wide; human disease; in vitro Assay; in vivo; individual variation; insight; inter-individual variation; microbial; receptor function; resistance gene; response; toxicant

Project Summary/Abstract The aryl hydrocarbon receptor (AHR) plays an essential role in the mechanisms of toxicity of numerous chemical contaminants, including chlorinated dioxins such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), some polychlorinated biphenyls (PCBs), and polynuclear aromatic hydrocarbons (PAHs). There is inter- individual variation in sensitivity to effects of these compounds, but the mechanisms are poorly understood. Variation in the human AHR sequence does not fully explain individual differences in response to AHR ligands, suggesting that other components of the AHR pathway are involved in controlling sensitivity. Recent genome- level research in fish populations with evolved resistance to PCBs, TCDD, and PAHs has identified the AHR- interacting protein (AIP) as a candidate resistance gene. AIP is an AHR chaperone that influences the stability and nuclear translocation of AHR, but its exact role is poorly defined. In humans, mutations in AIP predispose patients to familial isolated pituitary adenomas (FIPA), evidence that AIP sequence variation has functional consequences. Whether AIP variation alters the susceptibility to effects of AHR agonists in vivo is not known. The goal of this basic research is to elucidate the role of AIP and its sequence variants in controlling sensitivity to diverse AHR agonists, including environmental contaminants as well as natural AHR ligands. The central hypothesis is that variation at the AIP locus affects the interaction between AIP and AHR, leading to altered sensitivity to chemicals that cause toxicity and altered gene expression through the AHR. This hypothesis will be tested using complementary studies involving zebrafish (Danio rerio) in vivo and human cells in vitro. In Aim 1, AIP-null zebrafish generated using CRISPR-Cas9 genome-editing will be used to determine the role of AIP in controlling the sensitivity to developmental toxicity and altered gene expression caused by diverse AHR agonists in vivo. In Aim 2, the human liver cell line HepaRG will be used to investigate the molecular mechanisms by which AIP and its variants, including mutations associated with FIPA, affect AHR function. In Aim 3, targeted knock-in of AIP SNPs into zebrafish will be used to determine how variation in the AIP protein affects the sensitivity to diverse AHR ligands in vivo. The proposed research represents a unique opportunity to use insights from environmental exposures in wild fish populations along with mechanistic studies in human cells and engineered zebrafish embryos to understand fundamental mechanisms underlying individual differences in susceptibility to chemicals that act through the AHR. The studies address goals of the NIEHS 2018-2023 Strategic Plan, including basic research on molecular pathways involved in mediating effects of environmental exposures, research on developmental processes, and understanding mechanisms underlying individual susceptibility.

项目摘要/摘要 芳基烃受体（AHR）在众多毒性机制中起着至关重要的作用 化学污染物，包括氯化二恶英，例如2,3,7,8-四氯迪本佐-P-二恶英（TCDD）， 一些多氯联苯（PCB）和多核芳族碳氢化合物（PAHS）。有间 对这些化合物作用的敏感性的个体变化，但对这些机制的理解很少。 人类AHR序列的变化不能完全解释对AHR配体的响应的个体差异， 表明AHR途径的其他组件与控制灵敏度有关。最近的基因组 - 对PCB，TCDD和PAHS的进化抗性的鱼类种群的水平研究已经确定了AHR- 相互作用的蛋白（AIP）作为候选抗性基因。 AIP是一个影响的AHR伴侣 AHR的稳定性和核易位，但其确切作用的定义很差。在人类中，AIP中的突变 易孕症患者患有家族性孤立的垂体腺瘤（FIPA），证据表明AIP序列变异已有 功能后果。 AIP变化是否改变了对体内AHR激动剂的影响的敏感性是 不知道。这项基础研究的目的是阐明AIP及其序列变体的作用 控制对各种AHR激动剂的敏感性，包括环境污染物以及自然AHR 配体。中心假设是AIP基因座的变异会影响AIP和AHR之间的相互作用， 导致对引起毒性并通过AHR改变基因表达的化学物质的敏感性改变。 该假设将使用涉及斑马鱼（Danio Rerio）体内和人类的互补研究进行检验 细胞体外。在AIM 1中，使用CRISPR-CAS9基因组编辑产生的AIP-NULL斑马鱼将用于 确定AIP在控制对发育毒性敏感性和基因表达改变的敏感性中的作用 由体内的多种AHR激动剂引起。在AIM 2中，人肝细胞系HEPARG将用于调查 AIP及其变体（包括与FIPA相关的突变）的分子机制影响AHR 功能。在AIM 3中，将AIP SNP的目标敲入斑马鱼将用于确定如何变化 AIP蛋白会影响体内对不同AHR配体的敏感性。拟议的研究代表了一个独特的 使用野生鱼类种群环境暴露的见解以及机械的机会 人类细胞和工程斑马鱼胚胎的研究了解基本机制 通过AHR起作用的化学物质的敏感性的个体差异。研究解决了目标 NIEHS 2018-2023战略计划，包括有关介导的分子途径的基础研究 环境暴露的影响，对发展过程的研究以及理解机制的影响 基本的个人敏感性。