MicroRNA-199a-5p inhibition enhances the liver repopulation ability of human embryonic stem cell-derived hepatic cells
MicroRNA-199a-5p inhibition enhances the liver repopulation ability of human embryonic stem cell-derived hepatic cells
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DOI：
10.1016/j.jhep.2014.08.016
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发表时间：
2015-01-01
影响因子：
25.7
通讯作者：
Sharma, Amar Deep
中科院分区：
文献类型：
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作者：
Moebus, Selina;Yang, Dakai;Sharma, Amar Deep
背景与目的：目前人类胚胎干细胞（ESCs）的肝脏分化方案需要大量的改进。已经报道了MicroRNA（miRNAs）在肝脏发育期间调节肝细胞的命运，但它们在改善ESC向肝细胞分化方面的效用仍有待研究。因此，我们的目的是确定和分析肝源性miRNA的潜力，以提高肝细胞分化ESCs.Methods：通过miRNA的分析和体外筛选，我们确定了几个潜在的肝源性miRNA中的miR-199 a-5 p。在免疫缺陷延胡索酰乙酰乙酸水解酶敲除小鼠（Fah（-/-）/Rag 2（-/-）/Il 2 rg（-/-））的肝脏中进行miR-199 a-5 p抑制的肝细胞样细胞（HLC）的移植研究，以评估其体内肝脏再增殖潜力。结果：miRNA谱分析显示了20个保守的候选肝源性miRNA。通过miRNA筛选，发现仅HLC中的miR-199 a-5 p抑制能够增强小鼠以及人ESC的体外肝分化。人ESC衍生的HLC中的miR-199 a-5 p抑制增强了其在Fah（-/-）/Rag 2（-/-）/Il 2 rg（-/-）小鼠肝脏中的植入和再增殖能力。此外，我们确定SMARCA 4和MST 1作为miR-199 a-5 p的新靶点，可能有助于改善肝细胞生成和体内肝脏repopulation.Conclusions：我们的研究结果表明，在ES衍生的HLC中抑制miR-199 a-5 p可改善肝细胞分化。移植后，HLC能够移植并重新填充Fah（-/-）/Rag 2（-/-）/Il 2 rg（-/-）小鼠的肝脏。因此，我们的研究结果表明，miRNA调节可能是一种有前途的方法，从干细胞来源产生更成熟的HLC，用于治疗肝脏疾病。(C)2014年欧洲肝脏研究协会。Elsevier B. V.出版，保留所有权利。
Background & Aims: Current hepatic differentiation protocols for human embryonic stem cells (ESCs) require substantial improvements. MicroRNAs (miRNAs) have been reported to regulate hepatocyte cell fate during liver development, but their utility to improve hepatocyte differentiation from ESCs remains to be investigated. Therefore, our aim was to identify and to analyse hepatogenic miRNAs for their potential to improve hepatocyte differentiation from ESCs.Methods: By miRNA profiling and in vitro screening, we identified miR-199a-5p among several potential hepatogenic miRNAs. Transplantation studies of miR-199a-5p-inhibited hepatocytelike cells (HLCs) in the liver of immunodeficient fumarylacetoacetate hydrolase knockout mice (Fah(-/-)/Rag2(-/-)/Il2rg(-/-)) were performed to assess their in vivo liver repopulation potential. For target determination, western blot and luciferase reporter assay were carried out.Results: miRNA profiling revealed 20 conserved candidate hepatogenic miRNAs. By miRNA screening, only miR-199a-5p inhibition in HLCs was found to be able to enhance the in vitro hepatic differentiation of mouse as well as human ESCs. miR-199a-5p inhibition in human ESCs-derived HLCs enhanced their engraftment and repopulation capacity in the liver of Fah(-/-)/Rag2(-/-)/Il2rg(-/-) mice. Furthermore, we identified SMARCA4 and MST1 as novel targets of miR-199a-5p that may contribute to the improved hepatocyte generation and in vivo liver repopulation.Conclusions: Our findings demonstrate that miR-199a-5p inhibition in ES-derived HLCs leads to improved hepatocyte differentiation. Upon transplantation, HLCs were able to engraft and repopulate the liver of Fah(-/-)/Rag2(-/-)/Il2rg(-/-) mice. Thus, our findings suggest that miRNA modulation may serve as a promising approach to generate more mature HLCs from stem cell sources for the treatment of liver diseases. (C) 2014 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.