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。鲁棒 DNA加合物水平的测量可以为理解 DNA修复和允许评估人类暴露于遗传毒性化学品。我们推断一个预定的 选择反应监测（SRM）方法的DNA加合物分析将有助于高通量分析， DNA内收酶本申请的长期目标是开发一种靶向DNA内收体测定法， 用于定量评估由各种内源性和外源性DNA来源诱导的DNA加合物 有害物质，并采用该方法监测人类暴露于复杂的环境 污染物和研究DNA修复。本研究分为三个具体目标。在Aim中 #1，我们将建立SRM文库用于DNA内收体的靶向定量测量。在目标2中， 将全面分析暴露于颗粒物空气的小鼠肺组织中的DNA内收体 污染在目标3中，我们将评估Fe 2+和2-对烷基化DNA损伤的氧化代谢和修复， 酮戊二酸依赖性加氧酶。拟议研究的结果将提供科学的 社区的高通量平台，允许监测DNA内收体，并将提供新的 关于修复烷基化DNA损伤的见解。

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.