Department of Periodontology, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
Department of Cell Biology, School of Basic Medical Sciences, Peking University Stem Cell Research Center, Peking University, Beijing 100191, China.
Development. 2020 Dec 13;147(23):dev190298. doi: 10.1242/dev.190298.
E protein transcription factors are crucial for many cell fate decisions. However, the roles of E proteins in the germ-layer specification of human embryonic stem cells (hESCs) are poorly understood. We disrupted the gene locus to delete the E protein E2A in hESCs. E2A knockout (KO) hESCs retained key features of pluripotency, but displayed decreased neural ectoderm coupled with enhanced mesoendoderm outcomes. Genome-wide analyses showed that E2A directly regulates neural ectoderm and Nodal pathway genes. Accordingly, inhibition of Nodal or E2A overexpression partially rescued the neural ectoderm defect in E2A KO hESCs. Loss of E2A had little impact on the epigenetic landscape of hESCs, whereas E2A KO neural precursors displayed increased accessibility of the gene locus encoding the Nodal agonist CRIPTO. Double-deletion of both E2A and HEB () resulted in a more severe neural ectoderm defect. Therefore, this study reveals critical context-dependent functions for E2A in human neural ectoderm fate specification.
E 蛋白转录因子对于许多细胞命运决定至关重要。然而，E 蛋白在人类胚胎干细胞（hESC）的胚层特化中的作用还知之甚少。我们破坏了 基因座，以在 hESC 中删除 E 蛋白 E2A。E2A 敲除（KO）hESC 保留了多能性的关键特征，但表现出神经外胚层减少，同时增强中胚层结局。全基因组分析表明，E2A 直接调节神经外胚层和 Nodal 途径基因。因此，抑制 Nodal 或 E2A 过表达部分挽救了 E2A KO hESC 中的神经外胚层缺陷。E2A 的缺失对 hESC 的表观基因组景观几乎没有影响，而 E2A KO 神经前体细胞显示出编码 Nodal 激动剂 CRIPTO 的基因座的可及性增加。E2A 和 HEB（）的双重缺失导致更严重的神经外胚层缺陷。因此，这项研究揭示了 E2A 在人类神经外胚层命运特化中的关键上下文相关功能。
