Stable Isotope-Assisted Metabolomics for Deciphering Xenobiotic Metabolism in Mammalian Cell Culture
Stable Isotope-Assisted Metabolomics for Deciphering Xenobiotic Metabolism in Mammalian Cell Culture
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DOI：
10.1021/acschembio.9b01016
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发表时间：
2020-04-17
影响因子：
4
通讯作者：
Warth, Benedikt
中科院分区：
文献类型：
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作者：
Flasch, Mira;Bueschl, Christoph;Warth, Benedikt
外源性物质在环境中普遍存在，并通过I期和II期代谢在人体中进行修饰。液相色谱-高分辨质谱联用技术是研究这些生物转化产物的有力工具。我们提出了一个基于稳定同位素辅助代谢组学和生物信息学工具MetExtract H的工作流程，用于破译人类细胞产生的异生物质代谢物。其潜力被证明是通过调查的代谢脱氧雪腐镰刀菌烯醇（DON），丰富的食品污染物，在肝癌细胞系（HepG 2）和结肠癌模型（HT 29）。检测到的已知代谢物包括DON-3-硫酸盐、DON-10-磺酸盐2和DON-10-谷胱甘肽以及DON-半胱氨酸。首次证实了与氨基酸和抗生素的结合。该方法允许在组织培养中非靶向地阐明人类异生物质产物。它可以应用于其他研究领域，包括药物代谢，个性化医疗，麻烦研究和系统生物学，以更好地了解体外实验的相关性。
Xenobiotics are ubiquitous in the environment and modified in the human body by phase I and II metabolism. Liquid chromatography coupled to high resolution mass spectrometry is a powerful tool to investigate these biotransformation products. We present a workflow based on stable isotope-assisted metabolomics and the bioinformatics tool MetExtract H for deciphering xenobiotic metabolites produced by human cells. Its potential was demonstrated by the investigation of the metabolism of deoxynivalenol (DON), an abundant food contaminant, in a liver carcinoma cell line (HepG2) and a model for colon carcinoma (HT29). Detected known metabolites included DON-3-sulfate, DON-10-sulfonate 2, and DON-10-glutathione as well as DON-cysteine. Conjugation with amino acids and an antibiotic was confirmed for the first time. The approach allows the untargeted elucidation of human xenobiotic products in tissue culture. It may be applied to other fields of research including drug metabolism, personalized medicine, exposome research, and systems biology to better understand the relevance of in vitro experiments.