Wong Wing Tak, Matrone Gianfranco, Tian XiaoYu, Tomoiaga Simion Alin, Au Kin Fai, Meng Shu, Yamazoe Sayumi, Sieveking Daniel, Chen Kaifu, Burns David M, Chen James K, Blau Helen M, Cooke John P
Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States.
Department of Internal Medicine, University of Iowa, Iowa City, United States.
Elife. 2017 Mar 21;6:e23588. doi: 10.7554/eLife.23588.
We wish to identify determinants of endothelial lineage. Murine embryonic stem cells (mESC) were fused with human endothelial cells in stable, non-dividing, heterokaryons. Using RNA-seq, it is possible to discriminate between human and mouse transcripts in these chimeric heterokaryons. We observed a temporal pattern of gene expression in the ESCs of the heterokaryons that recapitulated ontogeny, with early mesodermal factors being expressed before mature endothelial genes. A set of transcriptional factors not known to be involved in endothelial development was upregulated, one of which was POU class 3 homeobox 2 (Pou3f2). We confirmed its importance in differentiation to endothelial lineage via loss- and gain-of-function (LOF and GOF). Its role in vascular development was validated in zebrafish embryos using morpholino oligonucleotides. These studies provide a systematic and mechanistic approach for identifying key regulators in directed differentiation of pluripotent stem cells to somatic cell lineages.
我们希望确定内皮细胞谱系的决定因素。将小鼠胚胎干细胞（mESC）与人类内皮细胞融合，形成稳定、不分裂的异核体。利用RNA测序，可以在这些嵌合异核体中区分人类和小鼠的转录本。我们观察到异核体胚胎干细胞中基因表达的时间模式重现了个体发育过程，早期中胚层因子在成熟内皮基因之前表达。一组未知参与内皮发育的转录因子被上调，其中之一是POU3类同源盒2（Pou3f2）。我们通过功能丧失和功能获得（LOF和GOF）证实了其在向内皮细胞谱系分化中的重要性。使用吗啉代寡核苷酸在斑马鱼胚胎中验证了其在血管发育中的作用。这些研究为识别多能干细胞向体细胞谱系定向分化中的关键调节因子提供了一种系统的机制方法。
