High temporal resolution proteome and phosphoproteome profiling of stem cell-derived hepatocyte development
High temporal resolution proteome and phosphoproteome profiling of stem cell-derived hepatocyte development
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DOI：
10.1016/j.celrep.2022.110604
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发表时间：
2022-03-29
期刊：
影响因子：
8.8
通讯作者：
Kuster，Bernhard
中科院分区：
文献类型：
--
作者：
Krumm，Johannes;Sekine，Keisuke;Kuster，Bernhard
原代人肝细胞被广泛用于评价药物的肝毒性，但其缺乏且培养要求高。干细胞衍生的肝细胞越来越多地被讨论为替代品。为了更好地了解诱导多能干细胞分化为肝细胞过程中的分子过程，我们采用定量蛋白质组学方法跟踪9，000种蛋白质，12，000个磷酸化位点和800个乙酰化位点随时间的表达。该分析揭示了阶段特异性标志物，即肝内胚层与未成熟肝细胞样细胞之间的主要分子开关，其影响例如，代谢、细胞周期、激酶活性和药物转运蛋白的表达。比较二维和三维肝细胞与胎儿和成人肝脏的蛋白质组表明体外模型的胎儿样状态和重要ADME/Tox蛋白的低表达。收集的数据使构建肝细胞发育的分子路线图成为未来研究的宝贵资源。
Primary human hepatocytes are widely used to evaluate liver toxicity of drugs, but they are scarce and demanding to culture. Stem cell-derived hepatocytes are increasingly discussed as alternatives. To obtain a better appreciation of the molecular processes during the differentiation of induced pluripotent stem cells into hepatocytes, we employ a quantitative proteomic approach to follow the expression of 9,000 proteins, 12,000 phosphorylation sites, and 800 acetylation sites over time. The analysis reveals stage-specific markers, a major molecular switch between hepatic endoderm versus immature hepatocyte-like cells impacting, e.g., metabolism, the cell cycle, kinase activity, and the expression of drug transporters. Comparing the proteomes of two- (2D) and three-dimensional (3D)-derived hepatocytes with fetal and adult liver indicates a fetal-like status of thein vitromodels and lower expression of important ADME/Tox proteins. The collective data enable constructing a molecular roadmap of hepatocyte development that serves as a valuable resource for future research.