Effects of Silymarin on the Metabolome

本行政补充申请是应PA-13-041“促进代谢组学研究合作活动”的要求提交的。这项请求的目的是促进一项新的合作，以表征对水飞蓟素(SM)的代谢反应，水飞蓟素(SM)是一种水飞蓟籽提取物，在许多动物模型中可以防止肝脏损伤和疾病进展，还可以在细胞培养中阻止丙型肝炎病毒(HCV)感染。增刊将汇集斯蒂芬·J·波利亚克博士、托马斯·O·梅茨博士和卡特里娜·沃特斯博士的实验室。波利亚克博士目前由NCCAM的R01AT006842资助，托马斯·O·梅茨博士是代谢组学方面的老牌专家，卡特里娜·沃特斯博士是生物信息学的专家。梅茨·S博士和沃特斯博士的实验室都在太平洋西北国家实验室。 父母赠款的主要假设是，SM衍生的天然产品以特定和多产的方式与哺乳动物生物分子相互作用，导致细胞中信号转导和基因表达的变化，从而保护肝脏。为了解决这一假设，我们使用了两种平行的方法来发现SM的作用机制。首先，我们正在使用微阵列分析来鉴定SM和SM衍生的纯化合物处理的肝细胞系和原代肝细胞培养物的转录变化。其次，利用光亲和点击化学，我们正在使用质谱学捕获和验证SM化合物的细胞靶标。基因芯片数据显示，SM在暴露后4-8小时内可诱导肝细胞基因表达的快速变化。有趣的是，许多变化都发生在与细胞增殖和代谢相关的基因上。这些和其他新出现的数据表明，SM正在诱导细胞快速新陈代谢重新编程。因此，我们的补充请求的目的是定义SM如何调节代谢组。我们将基于转录和蛋白质组学的R01扩展到包括代谢组学，这将使我们能够以前所未有的详细程度阐明SM等促进健康的天然产品是如何发挥其有益作用的。这一扩大的研究项目产生的新数据可能会为细胞如何对天然产品做出反应建立新的范式，这最终可能导致对多种人类疾病的天然产品治疗和/或药物开发机会的改进。

This Administrative Supplement request is submitted in response to PA-13-041 ¿Collaborative Activities to Promote Metabolomics Research¿. The intention of the request is to foster a new collaboration that will characterize the metabolomic response to silymarin (SM), an extract of milk thistle seeds, which prevents liver injury and disease progression in many animal models, and also blocks hepatitis C virus (HCV) infection in cell culture. The Supplement will bring together the laboratories of Dr. Stephen J. Polyak, currently funded by R01AT006842 from NCCAM, and Dr. Thomas O. Metz, an established expert in metabolomics, and Dr. Katrina Waters, an expert in bioinformatics. Both Drs. Metz¿s and Waters¿ laboratories are at Pacific Northwest National Laboratories (PNNL). The overarching hypothesis of the parent grant is that SM-derived natural products interact with mammalian biomolecules in a specific and productive manner to cause changes in signal transduction and gene expression in a cell to protect the liver. To address the hypothesis, we are using two parallel approaches to discover the mechanisms of action of SM. First, we are using microarray analysis to identify the transcriptional changes of liver cell lines and primary hepatocyte cultures treated with SM and SM-derived pure compounds. Second, using photoaffinity click chemistry, we are capturing and validating cellular targets of SM compounds using mass spectrometry. Microarray data indicate that SM induces rapid changes in hepatocellular gene expression within 4-8 hours after exposure. Intriguingly, many of the changes are in genes associated with cell proliferation and metabolism. These and other emerging data suggest that SM is inducing a rapid metabolic reprogramming of cells. Thus, the purpose of our supplement request is to define how SM modulates the metabolome. The extension of our transcriptional and proteomics-based R01 to include metabolomics will allow us to illuminate, to an unprecedented level of detail, exactly how health-promoting natural products such as SM exert their beneficial effects. The novel data emanating from this expanded research project may establish new paradigms for how cells respond to natural products, which may ultimately lead to refinements in natural product treatments and/or drug development opportunities for a multitude of human diseases.