Elsevier

Developmental Biology

Volume 384, Issue 2, 15 December 2013, Pages 155-165
Developmental Biology

The human constitutive androstane receptor promotes the differentiation and maturation of hepatic-like cells

https://doi.org/10.1016/j.ydbio.2013.10.012Get rights and content
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Highlights

  • Expression of CAR/NR1I3 in human embryonic stem cells drives maturation of hepatic-like cells.

  • In contrast, expression of PXR/NR1I2 was largely without effect in hepatic cell differentiation.

  • The results presented define a novel role for human CAR in hepatic lineage commitment.

Abstract

Expression of the constitutive androstane receptor (CAR, NR1I3) is enriched in the mature mammalian liver and increasingly recognized for its prominent role in regulating a myriad of processes including biotransformation, chemical transport, energy metabolism and lipid homeostasis. Previously, we demonstrated that CAR levels were markedly enhanced during the differentiation of hepatic-like cells derived from hESCs, prompting the hypothesis that CAR contributes a key functional role in directing human hepatogenesis. Here we demonstrate that over-expression of CAR in human embryonic stem cells (ESCs), transduced by a lentiviral vector, accelerates the maturation of hepatic-like cells, with CAR over-expressing cells exhibiting a 2.5-fold increase in albumin secretion by day 20 in culture differentiation, and significantly enhanced levels of mRNA expression of several liver-selective markers, including hepatic transcription factors, plasma proteins, biotransformation enzymes, and metabolic enzymes. CAR over-expressing cells also exhibited enhanced CITCO-inducible CYP3A7 enzymatic activity. Knockdown of CAR via siRNA attenuated the differentiation-dependent expression programs. In contrast, expression levels of the pregnane X receptor (PXR), a nuclear receptor most similar to CAR in primary sequence, were negligible in human fetal liver tissues or in the differentiating hESCs, and stable over-expression of PXR in hepatic-induced hESCs failed to enhance expression of hepatic phenotype markers. Together, these results define a novel role for human CAR in hepatic lineage commitment.

Keywords

CAR
PXR
Hepatic differentiation
Lentivirus
siRNA
hESCs
Drug metabolism

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