Center for Novel Biomarkers of Response

暴露生物学计划的最终目标是通过精确，准确和定量评估个人对环境压力源的暴露以及个人对这些压力源的反应来了解复杂疾病的发展和进展。人类发病率和死亡率的两个最重要的风险因素是暴露于香烟烟雾和肥胖，两者都与全身性慢性炎症和氧化应激相关。该中心的组织主题是，血浆中持久修饰蛋白的鉴定和验证将提供有关应激源，其作用模式和靶器官的具体信息。人类和动物项目的环境压力集中在主流和侧流香烟烟雾上，肥胖是一个令人困惑的生理因素。我们提出了一个综合的，多学科的中心，三个研究项目（人类，小鼠和传感器）和两个技术核心（蛋白质组学和ELISA微阵列），共同将有四个目标。目标1.发现活性氮和活性氧（RNS/ROS）修饰的肽使用MS/MS作为候选生物标志物。目标2.使用数据指导的MS验证RNS/ROS修饰的肽作为特异性生物特征。使用定制设计的夹心ELISA微阵列验证RNS/ROS修饰的蛋白质用作环境应激源的特异性生物标志物。目标4。 开发、测试和部署两个探测器系统，用于暴露以及RNS/ROS反应的特定和一般标记物。基于实验室的ELISA微阵列平台将用于提供高通量， 对数十种分析物进行多重分析。经过验证的生物标志物也将部署在原型，临床部署的检测器系统进行现场分析。该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），将支持在单个平台中测量暴露（可替宁）和反应的标记物，包括氧化应激和慢性炎症的特异性（修饰蛋白质）和通用标记物，如TNF-α。关键结果和可验证性。PNNL U 54中心将采用最先进的蛋白质组学和传感器技术， NIEHS具有告知反应生物标志物的作用模式数据库，用于在人类中测试和验证并通过小鼠中的平行研究获得信息的选定标志物的试剂，并部署在（a）基于实验室的ELISA微阵列平台上，以及（B）适用于大规模人类生物监测研究以评估基因与环境的相互作用的稳健的、临床可部署的基于纳米颗粒的传感器上。 U 54中心的研究结果还将提高科学界在人类和小鼠之间比较、对比和推断生物标志物的能力。

The ultimate goal of the Exposure Biology Program is to understand the development and progression of complex disease by precisely, 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, both assocaiated with systemic chronic inflammation and oxidative stress. The organizing theme the Center is that identification and validation of persistent modified proteins in plasma will provide specific information about the stressor, its mode of action, and the target organ. The environmental stress of the human and animal projects is focused on main-stream and side-stream cigarette smoke with obesity as a confounding physiological factor. We propose an integrated, multidisciplinary center with three research projects (human, mouse, and sensor) and two technology cores (proteomics and ELISA Microarray) which collectively will have four goals. Aim 1. Discover reactive nitrogen and reactive oxygen species (RNS/ROS) modified peptides using MS/MS as candidate biomarkers. Aim 2. Verification of RNS/ROS modified peptides as specific biosignatures using data-directed MS. Aim 3. Validation of RNS/ROS modified proteins for use as specific biomarkers for environmental stressors using custom-designed sandwich ELISA microarrays. Aim 4. Develop, test, and deploy two detector systems for exposure and for both specific and general markers of RNS/ROS response. A laboratory-based ELISA Microarray platform will be used to provide high-throughput, multiplexed analysis of dozens of analytes. The validated biomarkers will be also be deployed on a prototype, clinic-deployable detector system for on-site analysis. The sensor system is nanoparticle-based multiplexed Immunochromatographic / Electrochemical Biosensor (IEB) that will support the measurement of markers for exposure (cotinine) and response, both specific (modified proteins) and generic markers of oxidative stress and chronic inflammation, such as TNF-alpha, in a single platform. Critical Outcomes and Deliverables. The PNNL U54 Center will apply state-of-the-art proteomic and sensor technologies to provide NIEHS with a database of mode-of-action informing biomarkers of response, reagents for selected markers tested and validated in humans and informed by parallel studies in mice, and deployed on (a) a laboratory based ELISA Microarray platform, and (b) on a robust, clinic-deployable nanoparticle-based sensor suitable for use in large-scale human biomonitoring studies to evaluate the interaction of genes and the environment. The results of this U54 Center also will improve the scientific community's ability to compare, contrast and extrapolate biomarkers between humans and mice.