Chen Danyang, Fan Xiaonuo, Sun Ninghe, Wang Kai, Gong Liyan, Melero-Martin Juan M, Pu William T
Department of Cardiology, Boston Children's Hospital, Boston, MA, USA.
Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.
Nat Cardiovasc Res. 2025 Jun;4(6):689-709. doi: 10.1038/s44161-025-00660-y. Epub 2025 Jun 10.
Mechanisms of cell fate specification are central to developmental biology and regenerative medicine. ETV2 is a master regulator for the endothelial cell (EC) lineage specification. Here we study mechanisms by which ETV2 overexpression in human induced pluripotent stem-cell-derived mesodermal progenitors efficiently specifies ECs. We used CUT&RUN, scRNA-seq and scATAC-seq to characterize the molecular features of EC differentiation mediated by ETV2. We defined the scope of ETV2 pioneering activity and identified its direct downstream target genes. Induced ETV2 expression both directed specification of endothelial progenitors and suppressed acquisition of alternative fates. Functional screening and candidate validation revealed cofactors essential for efficient EC specification, including the transcriptional activator GABPA. Notably, the transcriptional repressor REST was also necessary for efficient EC specification. ETV2 recruited REST to repress non-EC lineage genes. Our study provides an unparalleled molecular analysis of EC specification at single-cell resolution and highlights the important role of pioneer factors to recruit repressors that suppress commitment to alternative lineages.
细胞命运特化机制是发育生物学和再生医学的核心。ETV2是内皮细胞（EC）谱系特化的主要调节因子。在此，我们研究了人类诱导多能干细胞衍生的中胚层祖细胞中ETV2过表达有效特化EC的机制。我们使用CUT&RUN、单细胞RNA测序（scRNA-seq）和单细胞染色质可及性测序（scATAC-seq）来表征ETV2介导的EC分化的分子特征。我们定义了ETV2先锋活性的范围，并鉴定了其直接下游靶基因。诱导的ETV2表达既指导内皮祖细胞的特化，又抑制其他命运的获得。功能筛选和候选验证揭示了高效EC特化所必需的辅助因子，包括转录激活因子GABPA。值得注意的是，转录抑制因子REST对高效EC特化也是必需的。ETV2招募REST来抑制非EC谱系基因。我们的研究提供了在单细胞分辨率下对EC特化的无与伦比的分子分析，并强调了先锋因子招募抑制因子以抑制向其他谱系分化的重要作用。
