Examination of transcriptional networks reveals an important role for TCFAP2C, SMARCA4, and EOMES in trophoblast stem cell maintenance

Abstract

Trophoblast stem cells (TS cells), derived from the trophectoderm (TE) of blastocysts, require transcription factors (TFs) and external signals (FGF4, INHBA/NODAL/TGFB1) for self-renewal. While many reports have focused on TF networks that regulate embryonic stem cell (ES cell) self-renewal and pluripotency, little is know about TF networks that regulate self-renewal in TS cells. To further understand transcriptional networks in TS cells, we used chromatin immunoprecipitation with DNA microarray hybridization (ChIP-chip) analysis to investigate targets of the TFs-TCFAP2C, EOMES, ETS2, and GATA3-and a chromatin remodeling factor, SMARCA4. We then evaluated the transcriptional states of target genes using transcriptome analysis and genome-wide analysis of histone H3 acetylation (AcH3). Our results describe previously unknown transcriptional networks in TS cells, including TF occupancy of genes involved in ES cell selfrenewal and pluripotency, co-occupancy of TCFAP2C, SMARCA4, and EOMES at a significant number of genes, and transcriptional regulatory circuitry within the five factors. Moreover, RNAi depletion of Tcfap2c, Smarca4, and Eomes transcripts resulted in a loss of normal colony morphology and down-regulation of TS cell-specific genes, suggesting an important role for TCFAP2C, SMARCA4, and EOMES in TS cell self-renewal. Through genome-wide mapping and global expression analysis of five TF target genes, our data provide a comprehensive analysis of transcriptional networks that regulate TS cell self-renewal.

Regulation of Trophoblast-Specific Factors by GATA2 and GATA3 in Bovine Trophoblast CT-1 Cells

“…It was recently reported that Gata3 and Cdx2 act in concert to induce the expression of common and independent target genes in the trophoblast lineage [35]. Moreover, transcription factors including GATA3 occupy regulatory regions of a number of target genes in TS cells [41]. GATA factors may interact alone or in combination with other transcription factors at the regulatory regions of trophoblast-specific factor genes, which in turn determine timing and degree of trophoblastspecific factor expression in trophoblast cells.…”

Diverse transcription factor binding features revealed by genome-wide ChIP-seq in C. elegans

“…S6). The signals over all targets were normalized and averaged to represent the overall distribution of the binding pattern for all targets of a given factor (Methods; Kidder and Palmer 2010). To assign a binding site to an annotated gene, we first collected coding and non-coding gene annotations from WormBase 170 and 200, respectively, while miRNAs were collected from miRBase v14 (Gerstein et al 2010).…”

BMP4-directed trophoblast differentiation of human embryonic stem cells is mediated through a ΔNp63+ cytotrophoblast stem cell state

“…However, to date, even the defining characteristics of such a human TSC remain controversial. Many genes have been designated, based on rodent studies, as required for establishment of the trophoblast lineage and/or maintenance of TSCs, including Cdx2, Elf5 and Eomes Niwa et al, 2005;Donnison et al, 2005;Ng et al, 2008;Russ et al, 2000;Kidder and Palmer, 2010). Some studies have identified the mRNA for these genes in human trophectoderm or primary trophoblast, and fewer have indisputably confirmed presence of the corresponding proteins in this compartment (Adjaye et al, 2005;Hemberger et al, 2010;Niakan and Eggan, 2013); even fewer studies have shown a definite role for these gene products in human trophoblast proliferation and/or differentiation.…”