Omics-Based Platform for Studying Chemical Toxicity Using Stem Cells
Omics-Based Platform for Studying Chemical Toxicity Using Stem Cells
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DOI：
10.1021/acs.jproteome.7b00693
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发表时间：
2018-01-01
影响因子：
4.4
通讯作者：
Wang, Daojing
中科院分区：
文献类型：
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作者：
Han, Yan;Zhao, Jinghua;Wang, Daojing
化学毒性测试和建模的新策略是使用体外基于人类细胞的测定结合定量高通量筛选（qHTS）技术，以确定分子机制并预测体内反应。干细胞比永生化细胞系更具有生理相关性，因为它们具有独特的增殖和分化潜力。我们建立了一个强大的双干细胞-双谱系检测系统，包括沿着骨发生的人间充质干细胞（hMSCs）和沿沿着肝发生的人诱导多能干细胞（hiPSC）。我们进行了LOPAC 1280的qHTS表型筛选，并鉴定了38个hMSC的初步命中。随后验证选定的命中数并确定其IC 50值，并使用蛋白质组学和生物信息学研究idarlavin和藜芦定治疗的机理。一般来说，hiPSC比hMSC对化学物质更敏感，分化的后代比其祖细胞更不敏感。我们发现，化学毒性取决于干细胞类型及其分化阶段。蛋白质组学鉴定和定量了两种干细胞的3000多种蛋白质。生物信息学鉴定细胞凋亡和G2/M为赋予艾达鲁肽毒性的主要途径。我们基于Omics的干细胞检测提供了对化学毒性的机理性见解，并可能有助于优先考虑化学品进行深入的毒理学评估。
The new strategy for chemical toxicity testing and modeling is to use in vitro human cell-based assays in conjunction with quantitative high-throughput screening (qHTS) technology, to identify molecular mechanisms and predict in vivo responses. Stem cells are more physiologically relevant than immortalized cell lines because of their unique proliferation and differentiation potentials. We established a robust two stem cells-two lineages assay system, encompassing human mesenchymal stem cells (hMSCs) along osteogenesis and human induced pluripotent stem cells (hiPSCs) along hepatogenesis. We performed qHTS phenotypic screening of LOPAC1280 and identified 38 preliminary hits for hMSCs. This was followed by validation of a selected number of hits and determination of their IC50 values and mechanistic studies of idarubicin and cantharidin treatments using proteomics and bioinformatics. In general, hiPSCs were more sensitive than hMSCs to chemicals, and differentiated progenies were less sensitive than their progenitors. We showed that chemical toxicity depends on both stem cell types and their differentiation stages. Proteomics identified and quantified over 3000 proteins for both stem cells. Bioinformatics identified apoptosis and G2/M as the top pathways conferring idarubicin toxicity. Our Omics-based assays of stem cells provide mechanistic insights into chemical toxicity and may help prioritize chemicals for in-depth toxicological evaluations.