MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, 5 Little France Drive, Edinburgh EH16 4UU, Scotland, UK.
Cell Stem Cell. 2013 May 2;12(5):531-45. doi: 10.1016/j.stem.2013.04.023.
Embryonic stem cell (ESC) pluripotency is governed by a gene regulatory network centered on the transcription factors Oct4 and Nanog. To date, robust self-renewing ESC states have only been obtained through the chemical inhibition of signaling pathways or enforced transgene expression. Here, we show that ESCs with reduced Oct4 expression resulting from heterozygosity also exhibit a stabilized pluripotent state. Despite having reduced Oct4 expression, Oct4(+/-) ESCs show increased genome-wide binding of Oct4, particularly at pluripotency-associated enhancers, homogeneous expression of pluripotency transcription factors, enhanced self-renewal efficiency, and delayed differentiation kinetics. Cells also exhibit increased Wnt expression, enhanced leukemia inhibitory factor (LIF) sensitivity, and reduced responsiveness to fibroblast growth factor. Although they are able to maintain pluripotency in the absence of bone morphogenetic protein, removal of LIF destabilizes pluripotency. Our findings suggest that cells with a reduced Oct4 concentration range are maintained in a robust pluripotent state and that the wild-type Oct4 concentration range enables effective differentiation.
胚胎干细胞(ESC)的多能性由以转录因子 Oct4 和 Nanog 为中心的基因调控网络控制。迄今为止,只有通过化学抑制信号通路或强制转基因表达才能获得强大的自我更新的 ESC 状态。在这里,我们表明,由于杂合性导致的 Oct4 表达减少的 ESC 也表现出稳定的多能状态。尽管 Oct4 表达减少,但 Oct4(+/-) ESC 显示出全基因组范围内 Oct4 的结合增加,特别是在多能相关增强子处,多能转录因子的同质表达,自我更新效率增强,分化动力学延迟。细胞还表现出 Wnt 表达增加、白血病抑制因子(LIF)敏感性增强和对成纤维细胞生长因子反应性降低。尽管它们能够在没有骨形态发生蛋白的情况下维持多能性,但去除 LIF 会破坏多能性。我们的发现表明,浓度降低的 Oct4 范围的细胞被维持在强大的多能状态,而野生型 Oct4 浓度范围则能够实现有效的分化。