HNF4A is essential for specification of hepatic progenitors from human pluripotent stem cells. Development

Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
Development (Impact Factor: 6.46). 08/2011; 138(19):4143-53. DOI: 10.1242/dev.062547
Source: PubMed


The availability of pluripotent stem cells offers the possibility of using such cells to model hepatic disease and development. With this in mind, we previously established a protocol that facilitates the differentiation of both human embryonic stem cells and induced pluripotent stem cells into cells that share many characteristics with hepatocytes. The use of highly defined culture conditions and the avoidance of feeder cells or embryoid bodies allowed synchronous and reproducible differentiation to occur. The differentiation towards a hepatocyte-like fate appeared to recapitulate many of the developmental stages normally associated with the formation of hepatocytes in vivo. In the current study, we addressed the feasibility of using human pluripotent stem cells to probe the molecular mechanisms underlying human hepatocyte differentiation. We demonstrate (1) that human embryonic stem cells express a number of mRNAs that characterize each stage in the differentiation process, (2) that gene expression can be efficiently depleted throughout the differentiation time course using shRNAs expressed from lentiviruses and (3) that the nuclear hormone receptor HNF4A is essential for specification of human hepatic progenitor cells by establishing the expression of the network of transcription factors that controls the onset of hepatocyte cell fate.

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Available from: Karim Si-Tayeb
    • "Quantification by RT-qPCR of a subset of mRNAs that are expressed in hepatic progenitor cells revealed a substantial reduction in the levels of HNF4A, Forkhead box A2 (FOXA2), HHEX, solute carrier family 35 (UDP-GlcA/UDP-GalNAc transporter) member D1 (SLC35D1), and phospholipase A2 group XIIB (PLA2G12B) mRNAs; however, there was no reduction in expression of GATA6 mRNA, which, like GATA4, is found in the endoderm prior to hepatic specification (Fig. 1C). We demonstrated previously that depletion of HNF4A in human pluripotent stem cells prevents hepatic specification (Delaforest et al. 2011). Given that the addition of PD161570 or PD173074 results in a dramatic reduction in expression of HNF4A as well as several other hepatic progenitor cell markers, we conclude that FGFR signaling acts upstream of HNF4A and is therefore essential for the production of hepatic progenitor cells from hiPSC-derived endoderm. "

    No preview · Article · Dec 2015 · Genes & Development
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    • "In the mouse embryo, the first expression of HNF4α is in the liver bud and gut tube at embryonic day 8.5, and then in the developing pancreas, kidney and in extra-embryonic visceral endoderm of the yolk sac [16]. The initial stages of hepatocyte differentiation are started by HNF4α that is accompanied by reduction in chromatin-mediated repression, resulting in a basal expression level of hepatocyte genes [17]. Many protocols are available for induction of the stem cells to differentiate into hepatocytes. "
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    ABSTRACT: The research on Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) from the umbilical cord suggests promising therapeutic use for hepatocyte replacement therapy. One of the highly conserved members of the nuclear receptor superfamily in the liver is hepatocyte nuclear factor-4α (HNF4α), involved in hepatocyte differentiation. The objectives of this study were to determine the effects of two- and threedimensional (2D and 3D) cultures of WJ-MSCs on hepatocyte differentiation. MSCs were isolated from human Wharton’s jelly, characterized by flow cytometry, and differentiated toward osteogenic and adipogenic lineage. WJ-MSCs were cultured in 2D collagen films and 3D collagen scaffolds in the presence of hepatogenic media with or without pre-treatment with fibroblast growth factor-4 (FGF4) for 21 days. The expression of HNF4α was evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). According to flow cytometry data, the cells isolated from Wharton’s jelly were shown to express MSC markers. HNF4α expression analysis revealed that pre-exposing the cells with FGF4 was more effective in hepatocyte differentiation. 3D cultures also improve the expression of HNF4α compared with 2D culture system. In conclusion, the combination of FGF4 and 3D culture improved hepatocyte differentiation. It seems 3D interaction of the cells improved the hepatogenesis.
    Full-text · Article · Jan 2015 · Romanian journal of morphology and embryology = Revue roumaine de morphologie et embryologie
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    • "In addition, HNF4a was found to be essential for the specification of hepatic progenitors from human pluripotent stem cells (iPSCs) [15]. The homeobox transcription factor Prox1 (prospero-related homeobox 1), an early specific marker during the development of liver and pancreas from the foregut endoderm, is a co-regulator of HNF4a and human liver receptor homolog-1 (hLRH-1) [16] [17]. "
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    ABSTRACT: Background & aims: Hepatocyte-like cells, differentiated from different stem cell sources, are considered to have a range of possible therapeutic applications, including drug discovery, metabolic disease modelling, and cell transplantation. However, little is known about how stem cells differentiate into mature and functional hepatocytes. Methods: Using transcriptomic screening, a transcription factor, liver X receptor α (NR1H3), was identified as increased during HepaRG cell hepatogenesis; this protein was also upregulated during embryonic stem cell and induced pluripotent stem cell differentiation. Results: Overexpressing NR1H3 in human HepaRG cells promoted hepatic maturation; the hepatocyte-like cells exhibited various functions associated with mature hepatocytes, including cytochrome P450 (CYP) enzyme activity, secretion of urea and albumin, upregulation of hepatic-specific transcripts and an increase in glycogen storage. Importantly, the NR1H3-derived hepatocyte-like cells were able to rescue lethal fulminant hepatic failure using a non-obese diabetic/severe combined immunodeficient mouse model. Conclusions: In this study, we found that NR1H3 accelerates hepatic differentiation through an HNF4α-dependent reciprocal network. This contributes to hepatogenesis and is therapeutically beneficial to liver disease.
    Preview · Article · Jul 2014 · Journal of Hepatology
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