Discovery of novel determinants of endothelial lineage using chimeric heterokaryons.
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.
Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States.
Wing Tak
WT
Wong
Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States.
Gianfranco
0000-0002-6064-0734
G
Matrone
Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States.
XiaoYu
X
Tian
Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States.
Simion Alin
SA
Tomoiaga
Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States.|Department of Internal Medicine, University of Iowa, Iowa City, United States.
Kin Fai
KF
Au
Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States.
Shu
S
Meng
Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, United States.
Sayumi
S
Yamazoe
Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, United States.
Daniel
D
Sieveking
Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States.
Kaifu
K
Chen
Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, United States.
David M
DM
Burns
Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, United States.
James K
JK
Chen
Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, United States.
Helen M
HM
Blau
Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States.
John P
0000-0003-0033-9138
JP
Cooke
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D002459:Cell Fusion; D053595:Embryonic Stem Cells / Q000502:physiology*; D042783:Endothelial Cells / Q000502:physiology*; D020869:Gene Expression Profiling; D006801:Humans; D051379:Mice; D015027:Zebrafish
Endothelial lineage; Heterokaryons; cell biology; developmental biology; human; mouse; nuclear reprogramming; pou3f2; stem cells; zebrafish