Protein Adducts as Measures of Exposure Throughout Childhood Development

 描述(由申请人提供)：胎儿、婴儿和儿童早期发育阶段非常容易受到环境危害的影响，在这些关键的敏感窗口期间暴露于有毒化学物质与儿童时期和整个一生中的疾病、残疾和不良健康有关。因此，在调查环境致病原因时，在生命早期接触的时间是一个关键因素。作为对人类基因组的补充，暴露组的概念--代表自受孕以来人类暴露的全部--在环境健康科学领域中变得越来越重要，用于研究疾病的病因。虽然目前还不可能使用单一实验来测量血液样本中的所有化学物质(类似于测量基因的全基因关联研究)，但在发展的关键时期可以研究重要的化学物质类别。一类重要的化学物质是亲电体，这是一种高活性的化合物，与绝大多数癌症和慢性病有关。亲电性既有外源性(如环境污染物和饮食)来源，也有内源性(如氧化应激和炎症)来源，因此反映了整个暴露-疾病连续体中范围广泛的重要分子。然而，由于亲电体在体内的寿命往往很短，通常不能在血液样本中直接测量它们。这促使使用蛋白质加成产物(加合物)作为生物标志物，以估计接触活性亲电化学品的情况，这反映了在蛋白质的停留时间(例如，几周至几个月)内接触短命亲电物质的综合情况。在这项提案中，我们将采用直接与NIEHS 2012-2017战略计划保持一致的两阶段方法。首先，我们将应用一种名为内收手术的新组学策略来绘制从出生前到21岁的早期生命暴露组图。使用这些实验的数据，以及从合作实验室收集的数据，我们将创建一个包含所有可检测到的HSA-Cys34的库 加合物。最后，我们将开发并验证一种高通量多路分析方法，用于定量干血样本中的靶向加合物-使用简单且微创的脚跟或手指刺伤在滤纸上收集的血滴。鉴于人类体内关于早期环境暴露及其对儿童健康影响的数据极为缺乏，这项提议为弥合这一关键的科学知识鸿沟提供了一个独特的机会。这项提案中开发的生物标记物方法将提供早期生命内收体组的第一张地图，并将为未来研究中调查疾病和障碍的环境原因提供具体工具。

 DESCRIPTION (provided by applicant): Periods of fetal, infant, and early childhood development are remarkably venerable to environmental hazards, and exposures to toxic chemicals during these critical windows of susceptibility have been linked with disease, disabilities, and adverse health in childhood and across the entire life span. Consequently, the timing of exposure during early-life is a critical factor to consider when investigating environmental causes of disease. As a complement to the human genome, the concept of the exposome - representing the totality of human exposures occurring from conception onward - has become increasingly salient in the field of environmental health sciences for investigating disease etiologies. While it is currently not possible to measure all chemicals in blood samples using single experiments (analogous to gene-wide association studies for measuring genes), important classes of chemicals can be investigated during critical period of development. One important class of chemicals is electrophiles, which are highly reactive compounds that have been linked to a vast majority of cancers and chronic diseases. Electrophiles have both exogenous (e.g., environmental pollutants and diet) and endogenous (e.g., oxidative stress and inflammation) sources, and thus reflect a broad spectrum of important molecules ranging across the entire exposure-disease continuum. However, because electrophiles tend to have very short life spans in vivo, they normally cannot be measured directly in blood samples. This has motivated the use of protein addition products (adducts) as biomarkers for estimating exposures to reactive electrophilic chemicals, which reflect an integration of exposure to short-lived electrophiles over the residence time of the protein (e.g., weeks to months). In this proposal we will apply a two-stage approach that directly aligns with the NIEHS 2012-2017 Strategic Plan. First, we will apply a new omics strategy called adductomics to map the early-life exposome from before birth onward to age 21. Using data from these experiments, as well as data collected from collaborating laboratories, we will then crate a library of all detectable HSA-Cys34 adducts. Finally, we will develop and validate a high-throughput muiltiplexed assay for quantifying panels of targeted adducts in dried blood samples - drops of blood collected on filter paper using a simple and minimally-invasive heel or finger prick. Given the overwhelming lack of human in vivo data on early-life environmental exposures and their impacts on children's health, this proposal offers a unique opportunity to bridge this critical gap in scientific knowledge. The biomarker approaches developed in this proposal will provide the first maps of the early-life adductome, and will provide specific tools for investigating environmental causes of diseases and disorders in future studies.