TCF/LEF regulation of the topologically associated domain ADI promotes mESCs to exit the pluripotent ground state

Cell Rep. 2021 Sep 14;36(11):109705. doi: 10.1016/j.celrep.2021.109705.

Abstract

Mouse embryonic stem cells (mESCs) can be maintained in vitro in defined N2B27 medium supplemented with two chemical inhibitors for GSK3 and MEK (2i) and the cytokine leukemia inhibitory factor (LIF), which act synergistically to promote self-renewal and pluripotency. Here, we find that genetic deletion of the four genes encoding the TCF/LEF transcription factors confers mESCs with the ability to self-renew in N2B27 medium alone. TCF/LEF quadruple knockout (qKO) mESCs display dysregulation of several genes, including Aire, Dnmt3l, and IcosL, located adjacent to each other within a topologically associated domain (TAD). Aire, Dnmt3l, and IcosL appear to be regulated by TCF/LEF in a β-catenin independent manner. Moreover, downregulation of Aire and Dnmt3l in wild-type mESCs mimics the loss of TCF/LEF and increases mESC survival in the absence of 2iL. Hence, this study identifies TCF/LEF effectors that mediate exit from the pluripotent state.

Keywords: TCF/LEF; Wnt signaling; mESC; pluripotency; β-catenin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIRE Protein
  • Animals
  • Benzamides / pharmacology
  • Cell Self Renewal* / drug effects
  • Culture Media / chemistry
  • Culture Media / pharmacology
  • DNA (Cytosine-5-)-Methyltransferases / antagonists & inhibitors
  • DNA (Cytosine-5-)-Methyltransferases / genetics
  • DNA (Cytosine-5-)-Methyltransferases / metabolism
  • Diphenylamine / analogs & derivatives
  • Diphenylamine / pharmacology
  • Down-Regulation / drug effects
  • Gene Editing
  • Hepatocyte Nuclear Factor 1-alpha / deficiency
  • Hepatocyte Nuclear Factor 1-alpha / genetics*
  • Hepatocyte Nuclear Factor 1-alpha / metabolism
  • Inducible T-Cell Co-Stimulator Ligand / antagonists & inhibitors
  • Inducible T-Cell Co-Stimulator Ligand / genetics
  • Inducible T-Cell Co-Stimulator Ligand / metabolism
  • Lymphoid Enhancer-Binding Factor 1 / deficiency
  • Lymphoid Enhancer-Binding Factor 1 / genetics*
  • Lymphoid Enhancer-Binding Factor 1 / metabolism
  • Mice
  • Mouse Embryonic Stem Cells / cytology
  • Mouse Embryonic Stem Cells / metabolism
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism
  • Pyridines / pharmacology
  • Pyrimidines / pharmacology
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Transcription Factor 7-Like 1 Protein / deficiency
  • Transcription Factor 7-Like 1 Protein / genetics*
  • Transcription Factor 7-Like 1 Protein / metabolism
  • Transcription Factor 7-Like 2 Protein / deficiency
  • Transcription Factor 7-Like 2 Protein / genetics*
  • Transcription Factor 7-Like 2 Protein / metabolism
  • Transcription Factors / antagonists & inhibitors
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • beta Catenin / deficiency
  • beta Catenin / genetics

Substances

  • Benzamides
  • Chir 99021
  • Culture Media
  • Hepatocyte Nuclear Factor 1-alpha
  • Hnf1a protein, mouse
  • Icosl protein, mouse
  • Inducible T-Cell Co-Stimulator Ligand
  • Lef1 protein, mouse
  • Lymphoid Enhancer-Binding Factor 1
  • Octamer Transcription Factor-3
  • Pou5f1 protein, mouse
  • Pyridines
  • Pyrimidines
  • RNA, Small Interfering
  • Tcf7l1 protein, mouse
  • Tcf7l2 protein, mouse
  • Transcription Factor 7-Like 1 Protein
  • Transcription Factor 7-Like 2 Protein
  • Transcription Factors
  • beta Catenin
  • mirdametinib
  • Diphenylamine
  • Dnmt3l protein, mouse
  • DNA (Cytosine-5-)-Methyltransferases

Grants and funding