Multiomics Analyses of HNF4α Protein Domain Function during Human Pluripotent Stem Cell Differentiation
Multiomics Analyses of HNF4α Protein Domain Function during Human Pluripotent Stem Cell Differentiation
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DOI：
10.1016/j.isci.2019.05.028
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发表时间：
2019-06-28
期刊：
影响因子：
5.8
通讯作者：
Hay, David C.
中科院分区：
文献类型：
--
作者：
Wang, Yu;Tatham, Michael H.;Hay, David C.
在哺乳动物发育过程中，肝脏分化是由汇聚在多个转录因子网络上的信号驱动的。众所周知，肝细胞核因子信号网络对肝细胞的规范和维持是必不可少的。在这项研究中，我们构建了肝细胞核因子-4α(HNF4α)的缺失和点突变，以精确评估人多能干细胞在肝细胞分化过程中蛋白结构域的功能。我们证明了核HNF4α对于肝脏前体细胞的指定是必不可少的，并且在HNF4α的小泛素样修饰物(SUMO)共识基序中引入点突变导致肝细胞分化中断。采用多组学方法，我们确定了细胞生物学中的关键缺陷，包括代谢功能障碍、底物黏附、三元酸循环通量、microRNA运输和mRNA加工。总之，基因组编辑和多组学分析的结合为HNF4α在多能干细胞进入肝脏谱系和肝细胞分化过程中的不同功能提供了有价值的见解。
During mammalian development, liver differentiation is driven by signals that converge on multiple transcription factor networks. The hepatocyte nuclear factor signaling network is known to be essential for hepatocyte specification and maintenance. In this study, we have generated deletion and point mutants of hepatocyte nuclear factor-4alpha (HNF4 alpha) to precisely evaluate the function of protein domains during hepatocyte specification from human pluripotent stem cells. We demonstrate that nuclear HNF4 alpha is essential for hepatic progenitor specification, and the introduction of point mutations in HNF4 alpha's Small Ubiquitin-like Modifier (SUMO) consensus motif leads to disrupted hepatocyte differentiation. Taking a multiomics approach, we identified key deficiencies in cell biology, which included dysfunctional metabolism, substrate adhesion, tricarboxylic acid cycle flux, microRNA transport, and mRNA processing. In summary, the combination of genome editing and multiomics analyses has provided valuable insight into the diverse functions of HNF4 alpha during pluripotent stem cell entry into the hepatic lineage and during hepatocellular differentiation.