Dax-1 Knockdown in Mouse Embryonic Stem Cells Induces Loss of Pluripotency and Multilineage Differentiation
Dax-1 Knockdown in Mouse Embryonic Stem Cells Induces Loss of Pluripotency and Multilineage Differentiation
复制标题
DOI：
10.1002/stem.78
复制
发表时间：
2009-01-01
期刊：
影响因子：
5.2
通讯作者：
Lalli, Enzo
中科院分区：
文献类型：
--
作者：
Khalfallah, Olfa;Rouleau, Matthieu;Lalli, Enzo
Dax-1（Nr 0 b1）是核激素受体超家族的孤儿成员，在肾上腺性腺发育和功能中具有关键作用。最近的研究还表明，Dax-1参与了控制胚胎干细胞（ES）多能性的转录网络。在这里，我们表明，Dax-1的表达是受分化治疗和药理学激活的β-连环蛋白依赖的转录在小鼠ES细胞。此外，Dax-1敲低诱导多系分化标志物的上调，并产生增强的分化和ES活力和增殖的缺陷。通过RNA干扰和转录组分析，我们已经确定了Dax-1在小鼠ES细胞中在敲低后24和48小时调控的基因。引人注目的是，这些基因中的绝大多数被上调，这表明Dax-1的普遍功能是在小鼠ES细胞中充当转录阻遏物，如通过使用Gal 4系统的实验所证实的。参与组织分化和增殖控制的基因在Dax-1调节的转录物中显著富集。这些数据表明，Dax-1是参与维持ES细胞多能性的分子回路中的重要元素，并且对理解生理（肾上腺）和临床（尤文瘤）环境中的干细胞功能具有意义，其中Dax-1分别在发育和发病机制中起关键作用。干细胞2009; 27：1529-1537
Dax-1 (Nr0b1) is an orphan member of the nuclear hormone receptor superfamily that has a key role in adrenogonadal development and function. Recent studies have also implicated Dax-1 in the transcriptional network controlling embryonic stem (ES) cell pluripotency. Here, we show that Dax-1 expression is affected by differentiating treatments and pharmacological activation of beta-catenin-dependent transcription in mouse ES cells. Furthermore, Dax-1 knockdown induced upregulation of multilineage differentiation markers, and produced enhanced differentiation and defects in ES viability and proliferation. Through RNA interference and transcriptome analysis, we have identified genes regulated by Dax-1 in mouse ES cells at 24 and 48 hours after knockdown. Strikingly, the great majority of these genes are upregulated, showing that the prevalent function of Dax-1 is to act as a transcriptional repressor in mouse ES cells, as confirmed by experiments using the Gal4 system. Genes involved in tissue differentiation and control of proliferation are significantly enriched among Dax-1 regulated transcripts. These data show that Dax-1 is an essential element in the molecular circuit involved in the maintenance of ES cell pluripotency and have implications for the understanding of stem cell function in both physiological (adrenal gland) and clinical (Ewing tumors) settings where Dax-1 plays a pivotal role in development and pathogenesis, respectively. STEM CELLS 2009; 27: 1529-1537