Elucidating Metabolic and Physicochemical Mechanisms of PAH Susceptibility in Toxicity Test Systems and Humans

项目摘要-Teguarden项目 缺乏对跨毒性测试系统敏感性的分子基础的了解是一个长期存在的问题 改进风险评估、改进SRP站点的清理决策和成功实现健康的障碍 减刑。我们的目标是开发变革性的、基于活性的蛋白质组探针和多模式 化学显微镜用来测量有助于差异的新陈代谢和分布过程 对多环芳烃暴露的敏感性。我们假设，可测量但通常被忽视的组织差异 分布和代谢能力激活和/或解毒多环芳烃有助于区分敏感性。 为了验证我们的假设，我们将完成三个目标：1.确定活性P450、谷胱甘肽-S- 代谢多环芳烃和多环芳烃代谢产物的转移酶和UDP-葡萄糖醛酸基转移酶。我们会 在组织样本中孵育基于活性的探针，利用我们的蛋白质组学丰富和鉴定活性酶 站台。2.建立一种测定多环芳烃代谢率和代谢敏感性的单一方法，方法如下： 将多环芳烃代谢率与全球酶活性测量联系起来。我们将孵化基于活动的探测器 组织进行标准多环芳烃代谢分析以确定底物特异性和代谢 活性酶的常量。3.确定测试系统和人体对暴露的敏感性 取决于多环芳烃的物理化学性质、组织成分和身体成分。我们将衡量 我们整个系统的组织组成，并测量和预测这些系统中多环芳烃的处置， 和人类。我们的研究成果直接增强了每个项目和核心的相关性和影响力 对利益相关者而言。我们的创新(ABPP和多模式化学显微镜)将开启新的科学 了解差异磁化率的两个关键分子决定因素的水平。这项工作将 通过以下方式满足对SRP的明确期望：a)证明“危险物质、其剂量、暴露 途径和模式生物在时间、流行和检测的背景下被考虑 B)确保“工作是与人类接触有关的背景”。 我们的研究结果将直接为超级基金网站的多环芳烃人类健康评估提供信息和改进。

PROJECT SUMMARY – TEEGUARDEN PROJECT Lack of understanding the molecular basis for susceptibility across toxicity test systems is a long-standing barrier to improved risk assessment, improved cleanup decisions for SRP sites, and successful health mitigations. Our objective is to develop transformative, activity-based proteomic probes and multi-modal chemical microscopy to measure the metabolic and distributional processes that contribute to differential susceptibility to PAH exposure. We hypothesize that measurable but commonly ignored differences in tissue distribution and metabolic capacity to activate and/or detoxify PAHs contributes to differential susceptibility. To test our hypothesis, we will complete three aims: 1. Identify the complete set of active P450, glutathione-S- transferase and UDP-glucuronosyltransferase enzymes that metabolize PAHs & PAH metabolites. We will incubate activity-based probes in tissue samples to enrich and identify active enzymes using our proteomics platform. 2. Develop a single assay to determine both PAH metabolic rates and metabolic susceptibility by relating PAH metabolic rates to global measures of enzyme activity. We will incubate activity-based probes in tissues while conducting standard PAH metabolism assays to determine substrate specificity and metabolic constants for active enzymes. 3. Determine how exposure susceptibility across test systems and humans depends upon PAH physicochemical properties, tissue composition, and body composition. We will measure tissue composition across our systems and measure and predict the disposition of PAHs across these systems, and humans. The products of our research directly enhance the relevance and impact of each project and core for stakeholders. Our innovations (ABPP and multi-modal chemical microscopy) will open new scientific horizons in the understanding of two key molecular determinants of differential susceptibility. The work will satisfy explicit expectations for SRP by a) demonstrating “that the hazardous substance, its dose, exposure pathway and model organisms are considered within the context of timing, prevalence, and detection of exposure” and by b) assuring that the “work is contextualized in terms of its relevance to human exposure.” Our research results will directly inform and improve human health assessment for PAHs at Superfund sites.