MicroRNA-199a-5p inhibition enhances the liver repopulation ability of human embryonic stem cell-derived hepatic cells

• Published in: Journal of hepatology Vol. 62; no. 1; pp. 101 - 110
• Main Authors: Möbus, Selina, Yang, Dakai, Yuan, Qinggong, Lüdtke, Timo H.-W, Balakrishnan, Asha, Sgodda, Malte, Rani, Bhavna, Kispert, Andreas, Araúzo-Bravo, Marcos J, Vogel, Arndt, Manns, Michael P, Ott, Michael, Cantz, Tobias, Sharma, Amar Deep
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.01.2015
• Subjects: Animals; Blotting, Western; Cell Differentiation; Cells, Cultured; Embryonic stem cells; Gastroenterology and Hepatology; Gene Expression Regulation; Hepatocyte differentiation; Hepatocytes - cytology; Hepatocytes - metabolism; Human Embryonic Stem Cells - cytology; Human Embryonic Stem Cells - metabolism; Humans; Liver repopulation; Liver Transplantation; Mice; Mice, Knockout; MicroRNA; MicroRNAs - antagonists & inhibitors; MicroRNAs - biosynthesis; MicroRNAs - genetics; Real-Time Polymerase Chain Reaction; RNA - genetics; mammalian STE20-like protein kinase 1/serine/threonine-protein kinase 4; NTBC; Hepatocyte differentiation; untranslated region; siRNA; small interfering RNA; MST1/STK4; Embryonic stem cells; SMARCA4; UTR; HLCs; 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione; miRNA; microRNA; SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 4; Liver repopulation; hepatocyte-like cells; MicroRNA
• ISSN: 0168-8278; 1600-0641
• DOI: 10.1016/j.jhep.2014.08.016

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 hepatocyte-like 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.