Oct4 and LIF/Stat3 additively induce Krüppel factors to sustain embryonic stem cell self-renewal.
Embryonic stem cell (ESC) pluripotency is dependent on an intrinsic gene regulatory network centered on Oct4. Propagation of the pluripotent state is stimulated by the cytokine leukemia inhibitory factor (LIF) acting through the transcriptional regulator Stat3. Here, we show that this extrinsic stimulus converges with the intrinsic circuitry in Krüppel-factor activation. Oct4 primarily induces Klf2 while LIF/Stat3 selectively enhances Klf4 expression. Overexpression of either factor reduces LIF dependence, but with quantitative and qualitative differences. Unlike Klf4, Klf2 increases ESC clonogenicity, maintains undifferentiated ESCs in the genetic absence of Stat3, and confers resistance to BMP-induced differentiation. ESCs expanded with Klf2 remain capable of contributing to adult chimeras. Postimplantation-embryo-derived EpiSCs lack both Klf2 and Klf4 and expression of either can reinstate naive pluripotency. These findings indicate that Oct4 and Stat3 intersect in directing expression of Klf transcriptional regulators with overlapping properties that additively reinforce ground-state ESC pluripotency, identity, and self-renewal.
Hall
John
J
Wellcome Trust Centre for Stem Cell Research, University of Cambridge, UK.
Guo
Ge
G
Wray
Jason
J
Eyres
Isobel
I
Nichols
Jennifer
J
Grotewold
Lars
L
Morfopoulou
Sofia
S
Humphreys
Peter
P
Mansfield
William
W
Walker
Rachael
R
Tomlinson
Simon
S
Smith
Austin
A
D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000378:metabolism; D054337:Cell Dedifferentiation; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D002678:Chimera; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism* / Q000473:pathology; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D053605:Leukemia Inhibitory Factor / Q000378:metabolism*; D051379:Mice; D046228:Microarray Analysis; D050814:Octamer Transcription Factor-3 / Q000378:metabolism*; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation; D000075746:Zinc Finger Protein Gli2
C525810:Bmp4 protein, mouse; D055415:Bone Morphogenetic Protein 4; C000618551:Gli2 protein, mouse; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D050796:STAT3 Transcription Factor; D000075746:Zinc Finger Protein Gli2