利用Hnf4α敲除的小鼠肝脏类器官探究发育模型的应用潜力  

Exploring the potential application of liver organoids of mouse as developmental models upon Hnf4α knockout

作　　者：张智玉 曹军 梁俊波 ZHANG Zhiyu;CAO Jun;LIANG Junbo(Department of Biochemistry and Molecular Biology,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所、北京协和医学院基础学院生物化学与分子生物学系,北京100005

出　　处：《基础医学与临床》2023年第5期724-732,共9页Basic and Clinical Medicine

基　　金：国家自然科学基金(82172887)。

摘　　要：目的探讨胆管类器官(ICO)分化模型用作肝脏发育模型的应用潜力。方法结合CRISPR/Cas9基因编辑技术和ICO分化模型,使用分别在扩增培养基、分化培养基及敲除肝细胞核因子4α(Hnf4α)后在分化培养基培养的类器官的全转录组测序数据分析ICO的转分化过程,使用公共测序数据集分析胎肝发育过程中的分化,并从基因表达的变化趋势、生物学功能,以Hnf4α为例的regulon调控关系这三个方面对两种分化过程进行比较。结果ICO转分化过程中基因表达的变化趋势与胎肝发育总体一致;转分化过程中表达上调的差异基因主要富集在与肝实质细胞生物学功能密切相关的代谢相关通路;ICO转分化过程中Hnf4α的下游靶基因与胎肝发育部分相似。结论ICO转分化模型可以部分模拟胎肝发育中的分化过程。Objective To explore the potential application of the intrahepatic cholangiocyte organoid(ICO)differentiation model to study liver development.Methods Combining CRISPR/Cas9 gene editing technology and ICO differentiation model,bulk RNA-seq data of the organoids cultured in expansion medium,differentiation medium,and differentiation medium after Hnf4αknockout were used to analyze the transdifferentiation process of ICO,and public single-cell transcriptome data sets were used to analyze the differentiation during fetal liver development.The two differentiation processes were compared in three aspects:the change of gene expression,biological function,and transcriptional regulation taking Hnf4αregulon as an example.Results The change of gene expression during ICO transdifferentiation was almost consistent with that in liver development.The up-regulated genes during transdifferentiation were mainly enriched in metabolism-related pathways closely associated biological functions of liver parenchymal cells.The downstream target genes of Hnf4αduring ICO transdifferentiation were overlapped with those in fetal liver development.Conclusions ICO transdifferentiation model can partially simulate the differentiation process of fetal liver development.

关 键 词：胆管类器官 转录组测序 分化 肝细胞核因子4α(Hnf4α) 

分 类 号：Q75[生物学—分子生物学]