A Targeted DNA Adductomics Approach for Analyzing > 100 DNA Adducts

摘要 内源性代谢和环境暴露可引起DNA加合物的形成。健壮 DNA加合物水平的测定可以为了解其发病机制提供重要的知识。 DNA修复，并允许评估人类接触遗传有毒化学物质。我们的理由是一个预定的 用于DNA加合物分析的选择反应监测(SRM)方法将有助于高通量分析 DNA加合物组。这一应用的长期目标是开发一种靶向dna加合试验。 用于定量评估各种内源和外源DNA诱导的DNA加合物 有害物质，并采用监测人类暴露在复杂环境中的方法 用于污染物和研究DNA修复。这项拟议的研究分为三个具体目标。在AIM 第一，我们将建立一个SRM文库，用于对DNA加合物组进行有针对性的定量测量。在目标2中，我们 将全面分析暴露在颗粒物空气中的小鼠肺组织中的DNA加合物组 污染。在目标3中，我们将评估Fe2+和2-Fe2+对烷化DNA损伤的氧化代谢和修复作用。 依赖草戊二酸的加氧酶。拟议的研究结果将提供科学的 具有高通量平台的社区，该平台允许监控DNA加合物组，并将负担 关于烷基化DNA损伤修复的见解。

ABSTRACT Endogenous metabolism and environmental exposure can give rise to DNA adduct formation. Robust measurement of the levels of DNA adducts can offer important knowledge for understanding the mechanisms of DNA repair and allow for assessment of human exposure to genotoxic chemicals. We reason that a scheduled selected-reaction monitoring (SRM) method for DNA adduct analysis will facilitate high-throughput analysis of the DNA adductome. The long-term objective of this application is to develop a targeted DNA adductomic assay for assessing quantitatively the DNA adducts induced by various endogenous and exogenous sources of DNA damaging agents, and to employ the method for monitoring human exposure to complicated environmental pollutants and for investigating DNA repair. The proposed research is organized into three Specific Aims. In Aim #1, we will establish an SRM library for targeted quantitative measurement of the DNA adductome. In Aim 2, we will profile comprehensively the DNA adductome in lung tissues of mice exposed to particulate matter air pollution. In Aim 3, we will assess the oxidative metabolism and repair of alkylated DNA lesions by Fe2+- and 2- oxoglutarate-dependent oxygenases. The outcome of the proposed research will provide the scientific community with a high-throughput platform that allows for monitoring the DNA adductome and will afford novel insights about the repair of alkylated DNA lesions.