MOUSE BIOMARKER DISCOVERY AND VALIDATION STUDIES

人类发病率和死亡率的两个最重要的可预防风险因素是接触 吸烟和肥胖，每一个都增加了患癌症、心血管或呼吸系统疾病的风险。 氧化应激可能是吸烟合并症发展的统一机制 以及这些危险因素与其他环境毒物和基因组的相互作用。 因此，本项目将建立在我们实验室在传感器开发、烟草烟雾 吸入毒理学、转录组学和蛋白质组学，在正常和 肥胖小鼠接触产生炎症和/或氧化应激的材料。研究将与 香烟烟雾（主流与侧流），以补充项目1中描述的人体研究， 随后暴露于炎性细菌剂、内毒素（脂多糖，LPS）和肺 和肝脏氧化剂百草枯和四氯化碳，以测试作用模式和组织依赖性 应答小鼠研究将有助于发现，验证，改进和验证候选药物。 氧化应激和炎症的蛋白质生物标志物，使用质谱蛋白质组学方法， 将被添加到PNNL的两个拟议的生物传感器平台-一个基于实验室的夹心ELISA微阵列 能够定量数十种蛋白质生物标志物（核心B）和基于临床可部署的纳米颗粒， 能够定量多种的多重免疫层析电化学生物传感器 暴露的生物标志物（例如环境化学品和/或其代谢物）和反应（蛋白质 氧化应激和炎症的生物标志物）（项目3）。小鼠研究将能够比较 在靶组织中的反应，这不能在人类中完成，与氧化的生物标志物 可以在通过渐进式微创方法收集的样品中定量的应激/炎症 (e.g.支气管肺泡灌洗液、血浆、唾液）以便于大规模部署生物传感器平台 人体生物监测研究。此外，该项目还提供了对下列答复的平行审查： 正常小鼠和肥胖小鼠的细胞，以确定定义这些关系的关键生物学途径。 环境因素之间的敏感性，从而创建一个重要的数据库， 全球基因和蛋白质谱分析，以进行小鼠到人类的外推。

Two of the most important preventable risk factors for human morbidity and mortality are exposure to cigarette smoke and obesity, each raising the risk of cancer, cardiovascular, or respiratory disease. Oxidative stress may be the unifying mechanism underlying the development of co-morbidities of smoking and obesity and for the interaction of these risk factors with other environmental toxicants and the genome. Thus, this project will build upon our Laboratory's experience in sensor development, tobacco smoke inhalation toxicology, transcriptomics, and proteomics, to conduct controlled exposure studies in normal and obese mice to materials that produce inflammation and/or oxidative stress. Studies will be conducted with cigarette smoke (mainstream vs. sidestream) to complement the human studies described in Project 1, followed by exposures to the inflammatory bacterial agent, endotoxin (lipopolysaccharide, LPS), and the lung and liver oxidants, paraquat and carbon tetrachloride, to test mode-of-action and tissue-dependent responses. The mouse studies will facilitate discovery, verification, refinement and validation of candidate protein biomarkers of oxidative stress and inflammation using mass spectrometry proteomic approaches that will be added to PNNL's two proposed biosensor platforms-a laboratory-based sandwich ELISA microarray capable of quantitating dozens of protein biomarkers (Core B) and an in-clinic deployable nanoparticlebased, multiplexed immunochromatographic electrochemical biosensor capable of quantitating multiple biomarkers of exposure (e.g. an environmental chemical and/or its metabolites) and response (protein biomarkers of oxidative stress and inflammation) (Project 3). The mouse studies will enable comparisons of responses in target tissues, which cannot be done in humans, with biomarkers of oxidative stress/inflammation that can be quantitated in samples collected by progressively less invasive approaches (e.g. bronchoalveolar lavage fluid, plasma, saliva) to facilitate biosensor platform deployment in large scale human biomonitoring studies. Furthermore, this project provides parallel examination of the responses in normal mice with those in obese mice to identify key biological pathways that define the relationships between the environmental factors contributing to susceptibility, thereby creating an important database of global gene and protein profiling to perform mouse to human extrapolations.