Discovery of novel determinants of endothelial lineage using chimeric heterokaryons

作者：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^*

来源：eLife, 2017, 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-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.

• 出版日期2017-3-21