title: Nanog-dependent feedback loops regulate murine embryonic stem cell heterogeneity.
abstract: A number of key regulators of mouse embryonic stem (ES) cell identity, including the transcription factor Nanog, show strong expression fluctuations at the single-cell level. The molecular basis for these fluctuations is unknown. Here we used a genetic complementation strategy to investigate expression changes during transient periods of Nanog downregulation. Employing an integrated approach that includes high-throughput single-cell transcriptional profiling and mathematical modelling, we found that early molecular changes subsequent to Nanog loss are stochastic and reversible. However, analysis also revealed that Nanog loss severely compromises the self-sustaining feedback structure of the ES cell regulatory network. Consequently, these nascent changes soon become consolidated to committed fate decisions in the prolonged absence of Nanog. Consistent with this, we found that exogenous regulation of Nanog-dependent feedback control mechanisms produced a more homogeneous ES cell population. Taken together our results indicate that Nanog-dependent feedback loops have a role in controlling both ES cell fate decisions and population variability.
authors.affiliation: Centre for Human Development, Stem Cells and Regeneration, Institute of Developmental Sciences, University of Southampton, Southampton SO17 1BJ, UK.
authors.forename: Ben D
authors.initials: BD
authors.lastname: MacArthur
authors.forename: Ana
authors.initials: A
authors.lastname: Sevilla
authors.forename: Michel
authors.initials: M
authors.lastname: Lenz
authors.forename: Franz-Josef
authors.initials: FJ
authors.lastname: Müller
authors.forename: Berhard M
authors.initials: BM
authors.lastname: Schuldt
authors.forename: Andreas A
authors.initials: AA
authors.lastname: Schuppert
authors.forename: Sonya J
authors.initials: SJ
authors.lastname: Ridden
authors.forename: Patrick S
authors.initials: PS
authors.lastname: Stumpf
authors.forename: Miguel
authors.initials: M
authors.lastname: Fidalgo
authors.forename: Avi
authors.initials: A
authors.lastname: Ma'ayan
authors.forename: Jianlong
authors.initials: J
authors.lastname: Wang
authors.forename: Ihor R
authors.initials: IR
authors.lastname: Lemischka
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D019295:Computational Biology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D005434:Flow Cytometry; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D000071317:Nanog Homeobox Protein; D060888:Real-Time Polymerase Chain Reaction; D059467:Transcriptome
chemical_list: D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse