Biomarkers of Exposure to Pulmonary Toxicants

肺部有毒物质暴露的生物标志物

• 批准号: 6900540
• 负责人: ALAN R BUCKPITT
• 金额: $ 15.88万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6900540
• 关键词: adduct; antioxidants; biomarker; cytotoxicity; environmental contamination; environmental exposure; laboratory mouse; laboratory rat; lung injury; naphthalenes; nitro compounds; particle; protein folding; respiratory epithelium; respiratory toxin; thiols

This application proposes to build on studies conducted during the past 5 years which have focused on two pulmonary cytotoxicants, naphthalene (NA) and 1-nitronaphthalene (NN). This work has: 1) demonstrated lung injury for NA at vapor concentrations well below the current occupational standard, 2) shown that the nasal epithelium is a susceptible target for both compounds, 3) identified many of the proteins adducted by reactive metabolites from NA and NN, and 4) demonstrated substantial quantitative differences in rodent vs nonhuman primate lung metabolism of these substrates. Overall, this work has contributed important data for current assessments of the potential human health hazards of these compounds. New methods have been developed which: 1) allow preservation of the lung for transcriptome analysis in well defined subcompartments, 2) allow selective sampling of proteins from airway epithelium facilitating analysis of changes in the airway proteome in response to toxicants and 3) improve analysis of alterations in protein expression by incorporation of a fluorescent internal standard. The foci of the studies proposed are to: 1) delineate differences in cytotoxic injury associated administration of respirable particle/chemical mixtures, 2) understand the importance of protein/nonprotein thiol oxidation in injury, 3) determine whether proteins which are adducted in the lung by reactive NA and NN metabolites are also adducted in susceptible nasal epithelium, 4) determine whether protein profiles and alterations in the metabolome present in nasal and bronchiolar lavage samples reflect injury to nasal and airway epithelium, 5) determine whether the nasal epithelium (and associated lavage sampling) can act as a legitimate surrogate for more distal parts of the lung and 6) delineate the importance of adducts with antioxidant enzymes and proteins involved in protein folding in cytotoxicity. These studies will provide important baseline data for work on the toxicology of 'real world' combustion mixtures. These particulars samples, many of which contain heavy metals, will be evaluated using nasal epithelium in vitro and in respiratory and renal tissue using proteomics approaches in vivo. The project depends upon close collaboration with the Thermal remediation project and will rely on the Analytical core for metabolomic, accelerator and protein mass spectrometry, and on the 'Omics and biostatistics core for proteome and transcriptome analysis.

本申请拟以过去 5 年进行的研究为基础，这些研究重点关注两种肺细胞毒剂：萘 (NA) 和 1-硝基萘 (NN)。这项工作：1) 证明了 NA 在远低于当前职业标准的蒸气浓度下会造成肺损伤，2) 表明鼻上皮是这两种化合物的敏感目标，3) 鉴定了许多由 NA 和 NA 的反应性代谢物加合的蛋白质。 NN和4)证明了这些底物的啮齿类动物与非人类灵长类动物肺代谢存在显着的定量差异。总体而言，这项工作为当前评估这些化合物对人类健康的潜在危害提供了重要数据。已经开发出新方法：1）允许在明确的子区室中保存肺部用于转录组分析，2）允许从气道上皮中选择性取样蛋白质，促进分析气道蛋白质组对毒物的反应变化，3）改进通过加入荧光内标来改变蛋白质表达。拟议研究的重点是：1）描绘与可吸入颗粒/化学混合物给药相关的细胞毒性损伤的差异，2）了解蛋白质/非蛋白质硫醇氧化在损伤中的重要性，3）确定在肺中加合的蛋白质是否反应性 NA 和 NN 代谢物也在易感鼻上皮中加合，4) 确定鼻腔和细支气管灌洗样品中存在的蛋白质谱和代谢组的变化是否反映对鼻和气道上皮的损伤，5) 确定鼻上皮（以及相关的灌洗取样）是否可以作为肺部更远端部分的合法替代物，6) 描述抗氧化酶和参与蛋白质折叠的蛋白质加合物的重要性在细胞毒性方面。这些研究将提供重要的 “现实世界”燃烧混合物毒理学工作的基线数据。这些特殊样本中许多含有重金属，将使用体外鼻上皮进行评估，并使用体内蛋白质组学方法评估呼吸和肾组织。该项目依赖于与热修复项目的密切合作，并将依赖代谢组学、加速器和蛋白质质谱分析核心，以及“组学和 用于蛋白质组和转录组分析的生物统计学核心。