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Inhibition of glycogen synthase kinase-3 alleviates Tcf3 repression of the pluripotency network and increases embryonic stem cell resistance to differentiation

Nature Cell Biology volume 13, pages 838–845 (2011)Cite this article

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An Erratum to this article was published on 02 May 2012

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Abstract

Self-renewal of rodent embryonic stem cells is enhanced by partial inhibition of glycogen synthase kinase-3 (Gsk3; refs 1, 2). This effect has variously been attributed to stimulation of Wnt signalling by β-catenin1, stabilization of Myc protein3 and global de-inhibition of anabolic processes4. Here we demonstrate that β-catenin is not necessary for embryonic stem cell identity or expansion, but its absence eliminates the self-renewal response to Gsk3 inhibition. Responsiveness is fully restored by truncated β-catenin lacking the carboxy-terminal transactivation domain5. However, requirement for Gsk3 inhibition is dictated by expression of T-cell factor 3 (Tcf3) and mediated by direct interaction with β-catenin. Tcf3 localizes to many pluripotency genes6 in embryonic stem cells. Our findings confirm that Tcf3 acts as a transcriptional repressor and reveal that β-catenin directly abrogates Tcf3 function. We conclude that Gsk3 inhibition stabilizes the embryonic stem cell state primarily by reducing repressive influence on the core pluripotency network.

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Figure 1: Suppression of Gsk3 mediates enhanced embryonic stem cell self-renewal but β-catenin is dispensable for embryonic stem cell maintenance.
Figure 2: β catΔ/− embryonic stem cells do not resist differentiation on Gsk3 inhibition.
Figure 3: β-catenin inhibits differentiation independently of its transcriptional activation domain.
Figure 4: β-catenin functions by abrogating Tcf3 repression.
Figure 5: Gsk3 inhibition relieves the core pluripotency network from repression by Tcf3 and complements Mek inhibition and/or Stat3 activation to stabilize embryonic stem cell self-renewal.

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  • 27 March 2012

    In the version initially published, the received date was incorrect. The correct received date is 20 May 2010.

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Acknowledgements

We thank N. Lyashenko and C. Hartmann for discussion and exchange of unpublished data. We also thank A. Martinez-Arias for comments on the manuscript. We are grateful to B. Merrill (University of Illinois at Chicago, USA) for generously providing Tcf3 mutant embryonic stem cells and to B. Doble (Stem Cell and Cancer Research Institute, McMaster University, Canada) and J. Woodgett (Samuel Lunenfeld Research Institute, Mount Sinai Hospital and University of Toronto, Canada) for Gsk3 mutant embryonic stem cells. We thank R. Grosschedl for the β-catenin ΔC construct. Gsk3 inhibitors, compounds A–G, were provided by Pfizer. R. Walker and P. Humphreys supported flow cytometry and imaging, respectively. The study was financially supported by the Biotechnology and Biological Sciences Research Council and the Medical Research Council of the United Kingdom, the Wellcome Trust and the European Commission FP7 project EuroSyStem. S.G-L. was supported by a CONACYT Studentship. A.S. is a Medical Research Council Professor.

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  1. Jason Wray, Sandra Gomez-Lopez & Dominik Eckardt

    Present address: Present addresses: Cancer Institute, University College London, Paul O’Gorman Building, 72 Huntley Street, London WC1E 6BT, UK (J.W.); Instituto de Fisiologı´a Celular, División de Neurociencias, UNAM, Circuito Exterior s/n, Ciudad Universitaria, México, DF 04510, Mexico (S.G-L.); Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, 51429 Bergisch Gladbach, Germany (D.E.),

Authors and Affiliations

  1. Wellcome Trust Centre for Stem Cell Research & Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK

    Jason Wray, Tüzer Kalkan, Sandra Gomez-Lopez & Austin Smith

  2. Max-Planck Institute of Immunobiology, Stubeweg 51, D-79108 Freiburg, Germany

    Dominik Eckardt & Rolf Kemler

  3. World Wide Medicinal Chemistry, Pfizer Ltd, Sandwich CT13 9NJ, UK

    Andrew Cook

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Contributions

J.W. carried out, analysed and interpreted experiments, T.K. created and validated the Rex1GFPd2 reporter, S.G-L. generated Cre–Ires–fluorescent protein plasmids, D.E. and R.K. generated floxed β-catenin embryonic stem cells, A.C. selected and provided Gsk3 inhibitors, and A.S. supervised the study and wrote the paper together with J.W.

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Correspondence to Austin Smith.

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Wray, J., Kalkan, T., Gomez-Lopez, S. et al. Inhibition of glycogen synthase kinase-3 alleviates Tcf3 repression of the pluripotency network and increases embryonic stem cell resistance to differentiation. Nat Cell Biol 13, 838–845 (2011). https://doi.org/10.1038/ncb2267

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