Effects of Silymarin on the Metabolome

本行政补充申请是为了响应PA-13-041而提交的。该请求的目的是促进一项新的合作，该合作将表征对水飞蓟素（SM）的代谢组学反应，水飞蓟素是一种奶蓟种子提取物，可预防许多动物模型中的肝损伤和疾病进展，并阻断细胞培养中的丙型肝炎病毒（HCV）感染。该补充将汇集斯蒂芬J.波利亚克博士的实验室，目前由NCCAM的R 01 AT 006842资助，和博士托马斯O。代谢组学专家梅斯和生物信息学专家卡特里娜沃茨博士。梅斯博士和沃茨博士的实验室都在太平洋西北国家实验室（PNNL）。 母基金的首要假设是SM衍生的天然产物以特定和有效的方式与哺乳动物生物分子相互作用，引起细胞中信号转导和基因表达的变化，以保护肝脏。为了解决这一假设，我们使用两种平行的方法来发现SM的作用机制。首先，我们使用微阵列分析来确定用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，使用光亲和点击化学，我们正在捕获和验证SM化合物的细胞目标，使用质谱。微阵列数据表明，SM在暴露后4-8小时内诱导肝细胞基因表达的快速变化。有趣的是，许多变化发生在与细胞增殖和代谢相关的基因中。这些和其他新出现的数据表明，SM正在诱导细胞的快速代谢重编程。因此，我们补充申请的目的是确定SM如何调节代谢组。我们基于转录和蛋白质组学的R 01扩展到包括代谢组学，将使我们能够以前所未有的细节水平阐明促进健康的天然产品（如SM）如何发挥其有益作用。从这个扩大的研究项目中产生的新数据可能会为细胞如何对天然产物做出反应建立新的范例，这可能最终导致天然产物治疗的改进和/或多种人类疾病的药物开发机会。

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.