Involvement of Hepatocyte Nuclear Factor 3 in Endoderm Differentiation of Embryonic Stem Cells

Abstract

The transcription factors of the hepatocyte nuclear factor 3 (HNF3) family, which are active in the liver, are expressed early during endoderm differentiation. To study their involvement in early murine development, we examined their role in embryonic stem (ES) cells. HNF3α or HNF3β mRNA transcripts were not detected in ES cells before differentiation, and only low levels of HNF3β mRNA were detected at a late stage of differentiation of ES cells to embryoid bodies (EB) (20 days after induction of differentiation). To examine the consequences of overexpressing HNF3α or -β in ES cells, we transfected the two genes into these cells and determined the levels of expression of tissue-specific genes during EB differentiation. Specifically, we examined expression of albumin, cystic fibrosis transmembrane conductance regulator (CFTR), phosphoenolpyruvate carboxykinase (PEPCK), α1-antitrypsin, transthyretin, ζ-globin, and neurofilament 68kd as markers for different cell lineages. Overexpression of HNF3β (and to a lesser extent of HNF3α) induced the expression of genes associated with endodermal lineage, namely, the genes for CFTR and albumin, but did not induce the expression of genes involved in late endoderm differentiation, such as the genes for PEPCK and α1-antitrypsin. Moreover, expression of HNF1β was highly induced in HNF3-overexpressing cells, while expression of HNF1α and HNF4 was only mildly induced in these cells. Therefore, HNF3α and -β seem to be involved in early endoderm differentiation of ES cells and together with other developmental factors are apparently needed for the induction of the endodermal lineage in vivo.