title: Reduced Oct4 expression directs a robust pluripotent state with distinct signaling activity and increased enhancer occupancy by Oct4 and Nanog.
abstract: Embryonic stem cell (ESC) pluripotency is governed by a gene regulatory network centered on the transcription factors Oct4 and Nanog. To date, robust self-renewing ESC states have only been obtained through the chemical inhibition of signaling pathways or enforced transgene expression. Here, we show that ESCs with reduced Oct4 expression resulting from heterozygosity also exhibit a stabilized pluripotent state. Despite having reduced Oct4 expression, Oct4(+/-) ESCs show increased genome-wide binding of Oct4, particularly at pluripotency-associated enhancers, homogeneous expression of pluripotency transcription factors, enhanced self-renewal efficiency, and delayed differentiation kinetics. Cells also exhibit increased Wnt expression, enhanced leukemia inhibitory factor (LIF) sensitivity, and reduced responsiveness to fibroblast growth factor. Although they are able to maintain pluripotency in the absence of bone morphogenetic protein, removal of LIF destabilizes pluripotency. Our findings suggest that cells with a reduced Oct4 concentration range are maintained in a robust pluripotent state and that the wild-type Oct4 concentration range enables effective differentiation.
authors: MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, 5 Little France Drive, Edinburgh EH16 4UU, Scotland, UK.
authors: Violetta
authors: V
authors: Karwacki-Neisius
authors: Jonathan
authors: J
authors: Göke
authors: Rodrigo
authors: R
authors: Osorno
authors: Florian
authors: F
authors: Halbritter
authors: Jia Hui
authors: JH
authors: Ng
authors: Andrea Y
authors: AY
authors: Weiße
authors: Frederick C K
authors: FC
authors: Wong
authors: Alessia
authors: A
authors: Gagliardi
authors: Nicholas P
authors: NP
authors: Mullin
authors: Nicola
authors: N
authors: Festuccia
authors: Douglas
authors: D
authors: Colby
authors: Simon R
authors: SR
authors: Tomlinson
authors: Huck-Hui
authors: HH
authors: Ng
authors: Ian
authors: I
authors: Chambers
mesh_terms: D001483:Base Sequence; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002999:Clone Cells; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D050814:Octamer Transcription Factor-3 / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D044967:Serum; D015398:Signal Transduction / Q000187:drug effects; D051153:Wnt Proteins / Q000378:metabolism
chemical_list: D019485:Bone Morphogenetic Proteins; D018398:Homeodomain Proteins; D050814:Octamer Transcription Factor-3; D051153:Wnt Proteins
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