We investigated the human induced pluripotent stem cells (hiPSCs) during a sequential in vitro step-by-step differentiation into hepatocyte-like cells (HLCs) using nanoCAGE, a method for promoters, transcription factors, and transcriptome analysis. Specific gene clusters reflected the different steps of the hepatic differentiation. The proliferation step was characterized by a typical cell cycle and DNA replication. The hepatic endoderm and the HLC steps were marked by a common signature including cell interactions with extracellular matrix (ECM), lipoproteins and hepatic biomarkers (such as albumin and alpha-fetoprotein). The specific HLC profile was characterized by important transcription factors such as HIF1A, JUN, MAF, KLF6, BMP4 and with a larger expression of genes related to Wnt signaling, extracellular matrix, lipid metabolism, urea cycle, drugs, and solute transporters. HLC profile was also characterized by the activation of upstream regulators such as HNF1A, MEIS2, NFIX, WRNIP1, SP4, TAL1. Their regulatory networks highlighted HNF4a as a bridge and linked them to important processes such as EMT-MET transitions, ECM remodeling and liver development pathways (HNF3, PPARA signaling, iron metabolism) along the different steps of differentiation.

中文翻译：

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在诱导多能干细胞逐步分化为肝细胞样细胞过程中转录因子及其调控作用的分析。

我们使用nanoCAGE，一种用于启动子，转录因子和转录组分析的方法，在依次体外逐步分化为类肝细胞（HLC）的过程中，研究了人类诱导的多能干细胞（hiPSC）。特定的基因簇反映了肝分化的不同步骤。增殖步骤的特征在于典型的细胞周期和DNA复制。肝内胚层和HLC步骤具有共同特征，包括细胞与细胞外基质（ECM），脂蛋白和肝生物标志物（如白蛋白和甲胎蛋白）的相互作用。特定的HLC图谱的特征在于重要的转录因子，例如HIF1A，JUN，MAF，KLF6，BMP4，以及与Wnt信号，细胞外基质，脂质代谢，尿素循环，药物和溶质转运蛋白。HLC曲线的特征还在于上游调节剂如HNF1A，MEIS2，NFIX，WRNIP1，SP4，TAL1的激活。他们的调节网络突出显示了HNF4a作为桥梁，并将它们与重要的过程（如EMT-MET转换，ECM重塑和肝脏发育途径（HNF3，PPARA信号传导，铁代谢））沿着不同的分化步骤联系在一起。