HUMAN BIOMARKER DISCOVERY AND VALIDATION STUDIES

暴露生物学计划的最终目标是了解的发展和进展， 复杂的疾病，通过准确和定量评估个人的暴露于环境 压力源和个人对这些压力源的反应。人类最重要的两个风险因素 发病率和死亡率与吸烟和肥胖有关。吸烟和肥胖都是 与全身慢性炎症和氧化应激有关。越来越多的证据表明， 由慢性炎症刺激引起的氧化应激是引起炎症的统一机制。 吸烟和肥胖共病的发展以及这些危险因素与 其他环境毒物和基因组。我们假设活性氮和活性氧 血浆中的RNS/ROS修饰蛋白将为香烟暴露提供重要的生物标志物 吸烟、肥胖的存在以及联合反应。项目1的总体目标是 鉴定、验证和确认人群中氧化应激血浆蛋白生物标志物， 四个目标：目标1。使用免疫组织化学鉴定RNS/ROS修饰的肽作为人血浆中的候选生物标志物 MS/MS。目标2将验证RNS/ROS修饰的肽作为120人血浆中的特异性生物特征 通过数据导向MS和FTICR-MS（核心A）分析样品。目标3：用于使用的CRNS/ROS修饰的蛋白质 作为特异性生物标志物，使用定制设计的夹心ELISA微阵列（核心B）。该确认将 包括抗体试剂开发和评价以及测定重现性和灵敏度（核心B）。 项目2将告知用于验证的蛋白质的选择。目标4。在实验室和 临床，用于暴露以及用于RNS/ROS反应的特异性和一般标志物的检测器系统（具有 项目3）。传感器系统是基于纳米颗粒的多路复用免疫色谱/电化学 生物传感器（IEB），支持测量暴露标记物（可替宁）和特定RNS/RNS 修饰的蛋白质和氧化应激和慢性炎症的通用反应标志物， 平台项目1将应用最先进的蛋白质组学分析的人血浆样本，从强大的，良好的 特征化的队列，为NIEHS提供候选生物标志物的数据库，以及用于选择的 暴露于香烟烟雾的标记物、肥胖的存在以及组合效应。生物标记物将 测试和验证一个包含500人的队列，并通过小鼠平行研究获得信息，以及 部署在强大的，在诊所部署纳米粒子为基础的传感器。

The ultimate goal of the Exposure Biology Program is to understand the development and progression of complex disease by accurately, and quantitatively assessing the individual's exposure to environmental stressors and the individual's responses to these stressors. Two of the most important risk factors for human morbidity and mortality are exposure to cigarette smoke and obesity. Smoking and obesity are each associated with systemic chronic inflammatory and oxidative stress. There is increasing evidence that the oxidative stress resulting from chronic inflammatory stimuli is the unifying mechanism underlying the development of co-morbidities in cigarette smoke and obesity and for the interaction of these risk factors with other environmental toxicants and the genome. We hypothesize that reactive nitrogen and reactive oxygen species (RNS/ROS) modified proteins in plasma will provide important biomarkers for exposure to cigarette smoke, presence of obesity, and the combined response. The overarching objective for Project 1 is to identify, verify, and validate plasma protein biomarkers of oxidative stress in a human population through four aims: Aim 1. Identify RNS/ROS-modified peptides as candidate biomarkers in human plasma using MS/MS. Aim 2 will Verify RNS/ROS-modified peptides as specific biosignatures in 120 human plasma samples by data-directed MS and FTICR-MS (Core A). Aim 3. Validate RNS/ROS-modified proteins for use as specific biomarkers using custom-designed sandwich ELISA microarrays (Core B). The validation will include development and evaluation of antibody reagents and assay reproducibility and sensitivity (Core B). The selection of proteins for validation will be informed by Project 2. Aim 4. Test in the laboratory and in the clinic, a detector system for exposure and for both specific and general markers of RNS/ROS response (with Project 3). The sensor system is nanoparticle-based multiplexed Immunochromatographic / Electrochemical Biosensor (IEB) that will support the measurement of markers for exposure (cotinine) and specific RNS/RNS modified proteins and generic response markers of oxidative stress and chronic inflammation on a single platform. Project 1 will apply state-of-the-art proteomic analysis of human plasma samples from robust, well characterized cohort to provide NIEHS a database of candidate biomarkers, and reagents for selected markers for exposure to cigarette smoke, presence of obesity, and the combined effect. The biomarkers will be tested and validated a cohort containing 500 humans and informed by parallel studies in mice, and deployed on robust, in-clinic deployable nanoparticle-based sensors.