ZFIN ID: ZDB-PUB-170323-19
Discovery of novel determinants of endothelial lineage using chimeric heterokaryons
Wong, W.T., Matrone, G., Tian, X., Tomoiaga, S.A., Au, K.F., Meng, S., Yamazoe, S., Sieveking, D., Chen, K., Burns, D.M., Chen, J.K., Blau, H.M., Cooke, J.P.
Date: 2017
Source: eLIFE   6: (Journal)
Registered Authors: Chen, James K.
Keywords: cell biology, developmental biology, human, mouse, stem cells, zebrafish
MeSH Terms:
  • Animals
  • Cell Differentiation/genetics*
  • Cell Fusion
  • Embryonic Stem Cells/physiology*
  • Endothelial Cells/physiology*
  • Gene Expression Profiling
  • Humans
  • Mice
  • Zebrafish
PubMed: 28323620 Full text @ Elife
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.