Project 4: Urinary Protein Biomarkers for Assessing the Potential Toxicity

项目 4：用于评估潜在毒性的尿蛋白生物标志物

• 批准号: 8659362
• 负责人: BRUCE D HAMMOCK
• 金额: $ 15.14万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8659362
• 关键词: Acetylcysteine; Acute; Address; Air; Air Force Personnel; Aluminum; American; Animal Model; Animals; Antibodies; Aromatic Hydrocarbons; Benign; Binding; Biological Assay; Biological Markers; Biological Models; Breathing; Carcinogens; Cells; Chemical Models; Chemicals; Chronic; Coal; Collaborations; Contracts; Core Facility; Cytochrome P450; Data; Deposition; Disease; Dose; Engineering; Epidemiology; Epithelial Cells; Event; Exposure to; Female; Food; Fossil Fuels; Genes; Genetic Polymorphism; Glucuronides; Goals; Hazardous Substances; Health; Hepatic; Human; Hyperplasia; In Situ; Individual; Inhalation Exposure; Injury; Inorganic Sulfates; Institutes; Label; Lethal Dose 50; Letters; Link; Liver; Lung; Lung diseases; Macaca mulatta; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Measures; Metabolic Activation; Metabolism; Methods; Military Personnel; Mixed Function Oxygenases; Modeling; Modification; Molecular; Molecular Profiling; Monitor; Monkeys; Morbidity - disease rate; Mus; NADPH-Ferrihemoprotein Reductase; Naphthalene; Naphthols; Naphthoquinones; Nasal Epithelium; Nasal cavity; National Health and Nutrition Examination Survey; Necrosis; Nose; Occupational; Oils; Olfactory Epithelial Cell; Olfactory Epithelium; Oxides; Peptides; Petroleum; Photons; Plants; Population; Primates; Procedures; Process; Protein Binding; Proteins; Proteomics; Rattus; Recovery; Respiratory System; Respiratory tract structure; Risk; Risk Assessment; Rodent; Rodent Model; Route; Sampling; Services; Site; Soil; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Superfund; Surveys; System; Testing; Tissues; Toxic effect; Toxicology; Uncertainty; Unspecified or Sulfate Ion Sulfates; Urine; Water; Western Blotting; Work; Workplace; accelerator mass spectrometry; adduct; airway epithelium; base; cytotoxic; cytotoxicity; design; exposed human population; indexing; injury and repair; liquid chromatography mass spectrometry; lung injury; male; mortality; neoplastic; protein metabolite; respiratory; response; sensor; toxicant; urinary; wasting

Lung diseases are a significant cause of morbidity and mortality in the US population. Exposure to chemicals in air, water and food contributes to these diseases. A number of chemicals undergo bioactivation to produce selective lung injury in rodents. This project focuses on naphthalene (NA) and 1-nitronaphthalene (NN), which are present in superfund waste sites and which represent a class of environmentally important compounds. Both chemicals undergo metabolism to electrophilic intermediates which become bound covalently to proteins in target cells. Protein binding is a key component of cytotoxicity. We have demonstrated that the proteins targeted are similar in rodents and primate nasal epithelium. This underscores the need to develop biomarkers which are tightly tied to toxicity. The central hypothesis is: /the formation of key protein adducts with reactive metabolites of NA/NN in target respiratory tissue is causally related to cytotoxicity; measurement of adducted peptides/proteins in urine and nasal brushings provides a highly sensitive molecular signature of exposure and effect./ The proposed work builds on recent findings showing 1) high specific activity adducts in urine of NA-treated mice, and 2) numerous identified protein adducts in lungs and nasal cavity of mice, rats and monkeys. The proposed studies will utilize antibodies from project 3 to trap and concentrate adducted peptides from the urine and to evaluate adduct levels in animal model systems. These approaches will be validated for their ability to assess adduct levels following exposures to small amounts of 14C-NA by accelerator mass spectrometry (Core A), will utilize the mass spectrometry facilities of cores A and B for analysis of adducted peptides and will depend heavily on the proteomics services offered by Core B. Overall, these studies are expected to: 1) yield fundamental information on how adducted proteins, formed in respiratory tissues, are handled in the whole animal, 2) explore concentration-response relationships at environmentally relevant concentrations, 3) provide methods which can be applied to exposed human populations, which will be useful-with modification-for assessing risk of other metabolically activated chemicals and 4) provide methods which can clarify the importance of genetic polymorphisms in genes responsible for the biodisposition of chemicals like NA and NN.

肺部疾病是美国人口发病率和死亡率的重要原因。暴露于空气，水和食物中的化学物质会导致这些疾病。许多化学物质会经历生物活化，以在啮齿动物中产生选择性肺损伤。该项目侧重于萘（NA）和1-硝基萘（NN），它们存在于超级基金废物站点中，它们代表了一类具有环境重要化合物的化合物。两种化学物质都对亲电中间体的新陈代谢构成，这些中间体与靶细胞中的蛋白质共价结合。蛋白质结合是细胞毒性的关键组成部分。我们已经证明，靶向的蛋白质在啮齿动物和灵长类动物鼻上皮相似。这强调了开发与毒性紧密相关的生物标志物的需求。中心假设是： /在靶呼吸组织中Na /Nn的反应性代谢物的关键蛋白加合物的形成与细胞毒性有因果关系；尿液和鼻刷中加合肽/蛋白质的测量提供了高度敏感的暴露和效果的分子特征。/拟议的工作构建的基于最新发现，显示1）NA处理的小鼠尿液中的高特异性活性加合物，以及2）众多鉴定的尿液小鼠，大鼠和猴子的肺和鼻腔中的蛋白质加合物。拟议的研究将利用从项目3到陷阱的抗体，并从尿液中浓缩肽中的肽并评估动物模型系统中的加合物水平。这些方法将通过加速器质谱法（核心A）暴露于少量14C-NA后评估加合物水平的能力进行验证（核心A） 核A和B的光谱学设施用于分析加合肽，并将在很大程度上取决于核心B提供的蛋白质组学服务。总体而言，这些研究预计：1）关于在呼吸道组织中形成的加合物蛋白如何处理的基本信息。在整个动物中，2）在环境相关浓度下探索浓度 - 响应关系，3）提供可以应用于暴露的人类种群的方法，这将是有用的，可以在评估其他代谢化学品的风险中进行修改，4）提供可以阐明遗传多态性在负责Na和Nn等化学物质生物斑层的基因中的重要性的方法。