干细胞转分化肝细胞被认为是最具前景的原代肝细胞替代细胞源，但转分化机制不清，严重阻碍临床转化应用。课题组前期已创建基于人骨髓间充质干细胞（hBMSC）移植的人肝嵌合小鼠，发现植入干细胞3天内完成转分化，经增殖后进入功能成熟期，移植后2天内特异性高表达基因与肝细胞功能密切相关，其中DNMBP参与调控转分化肝细胞的成熟。本研究拟在上述基础上，构建DNMBP过表达和敲除的hBMSC，分别进行体外肝向诱导分化和体内肝衰竭微环境下转分化研究，通过人肝细胞特异性基因表达、人白蛋白分泌、尿素合成、安定代谢能力等观察DNMBP表达变化对转分化细胞功能成熟的影响；利用转录组测序和课题组自主研发的多组学功能关联分析技术，揭示转分化不同阶段分子特征，寻找DNMBP调控肝细胞功能的信号通路及关键分子，阐明DNMBP对转分化肝细胞成熟的调控机制。为最终使用转分化肝细胞进行再生医学研究和临床应用提供理论基础。

Stem cell differentiated hepatocyte is a promising alternative of primary human hepatocyte, however the differentiation mechanism is still unclear, which limits the further clinical translation of the differentiated hepatocyte. During our previous research, we have established a liver humanized mouse model via transplantation of human bone marrow mesenchymal stem cell (hBMSC), and found that the transplanted hBMSC differentiated into hepatocyte like cells within three days, then these cells went through proliferation and maturation. Genes that were significantly differentially upregulated within two days were related with the regulation of liver function, among them DNMBP was found to be involved in the regulation of hepatocyte maturation. In this study, we will overexpress and knock down the gene expression of DNMBP in hBMSC, then perform the hepatogenic induction in vitro and in vivo, and evaluate the effect of DNMBP on the maturation of hBMSC-differentiated hepatocytes through the detections of human hepatocyte specific gene expressions, human albumin secretion, urea synthesis and metabolism of diazepam. mRNA sequencing and our in-house multi-omics cross-validation and network analysis will be used to analyze the transcriptome profiling at different differentiation stage after management of DNMBP, and to discover the signaling pathways and key genes that were regulated by DNMBP, finally clarify the mechanism of DNMBP on the maturation of hBMSC-derived hepatocytes. This research will provide theoretical foundation for the translation of stem cell-derived hepatocytes in regeneration medicine and clinical use.