Loss of the Otx2-Binding Site in the Nanog Promoter Affects the Integrity of Embryonic Stem Cell Subtypes and Specification of Inner Cell Mass-Derived Epiblast

Cell Rep. 2016 Jun 21;15(12):2651-64. doi: 10.1016/j.celrep.2016.05.041. Epub 2016 Jun 9.

Abstract

Mouse embryonic stem cells (ESCs) and the inner cell mass (ICM)-derived epiblast exhibit naive pluripotency. ESC-derived epiblast stem cells (EpiSCs) and the postimplantation epiblast exhibit primed pluripotency. Although core pluripotency factors are well-characterized, additional regulators, including Otx2, recently have been shown to function during the transition from naive to primed pluripotency. Here we uncover a role for Otx2 in the control of the naive pluripotent state. We analyzed Otx2-binding activity in ESCs and EpiSCs and identified Nanog, Oct4, and Sox2 as direct targets. To unravel the Otx2 transcriptional network, we targeted the strongest Otx2-binding site in the Nanog promoter, finding that this site modulates the size of specific ESC-subtype compartments in cultured cells and promotes Nanog expression in vivo, predisposing ICM differentiation to epiblast. Otx2-mediated Nanog regulation thus contributes to the integrity of the ESC state and cell lineage specification in preimplantation development.

Keywords: Nanog; Otx2; embryonic stem cells; pluripotency; preimplantation epiblast.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Blastocyst / cytology*
  • Blastocyst / drug effects
  • Blastocyst / metabolism
  • Cell Compartmentation / drug effects
  • Cell Differentiation / drug effects
  • Cell Lineage / drug effects
  • Cell Proliferation / drug effects
  • Chimera / metabolism
  • Embryonic Development / drug effects
  • Embryonic Development / genetics
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Endoderm / cytology
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental / drug effects
  • Germ Layers / cytology*
  • Germ Layers / drug effects
  • Germ Layers / metabolism
  • Leukemia Inhibitory Factor / pharmacology
  • Mesoderm / cytology
  • Mice
  • Mutation / genetics
  • Nanog Homeobox Protein / genetics*
  • Nanog Homeobox Protein / metabolism
  • Otx Transcription Factors / genetics
  • Otx Transcription Factors / metabolism*
  • Promoter Regions, Genetic / genetics*
  • Protein Binding / drug effects

Substances

  • Leukemia Inhibitory Factor
  • Nanog Homeobox Protein
  • Otx Transcription Factors
  • Otx2 protein, mouse