EMD Serono Research Institute, Inc., Rockland, MA 02370, USA.
Genome Res. 2010 Apr;20(4):458-72. doi: 10.1101/gr.101469.109. Epub 2010 Feb 22.
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 self-renewal 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.
滋养层干细胞（TS 细胞）来源于囊胚的滋养外胚层（TE），需要转录因子（TFs）和外部信号（FGF4、INHBA/NODAL/TGFB1）来自我更新。虽然许多报告都集中在调节胚胎干细胞（ES 细胞）自我更新和多能性的 TF 网络上，但对于调节 TS 细胞自我更新的 TF 网络知之甚少。为了进一步了解 TS 细胞中的转录网络，我们使用染色质免疫沉淀与 DNA 微阵列杂交（ChIP-chip）分析来研究 TF-TCFAP2C、EOMES、ETS2 和 GATA3 的靶标，以及一个染色质重塑因子 SMARCA4。然后，我们使用转录组分析和组蛋白 H3 乙酰化（AcH3）的全基因组分析来评估靶基因的转录状态。我们的结果描述了 TS 细胞中以前未知的转录网络，包括 ES 细胞自我更新和多能性相关基因的 TF 占据、TCFAP2C、SMARCA4 和 EOMES 在大量基因上的共同占据，以及五个因子内的转录调控电路。此外，RNAi 耗尽 Tcfap2c、Smarca4 和 Eomes 转录本导致正常集落形态丧失和 TS 细胞特异性基因下调，表明 TCFAP2C、SMARCA4 和 EOMES 在 TS 细胞自我更新中具有重要作用。通过对五个 TF 靶基因的全基因组作图和全局表达分析，我们的数据提供了对调节 TS 细胞自我更新的转录网络的全面分析。
