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FOXO1 is an essential regulator of pluripotency in human embryonic stem cells

Nature Cell Biology volume 13, pages 10921099 (2011) | Download Citation

Abstract

Pluripotency of embryonic stem cells (ESCs) is defined by their ability to differentiate into three germ layers and derivative cell types1,2,3 and is established by an interactive network of proteins including OCT4 (also known as POU5F1; ref. 4), NANOG (refs 5, 6), SOX2 (ref. 7) and their binding partners. The forkhead box O (FoxO) transcription factors are evolutionarily conserved regulators of longevity and stress response whose function is inhibited by AKT protein kinase. FoxO proteins are required for the maintenance of somatic and cancer stem cells8,9,10,11,12,13; however, their function in ESCs is unknown. We show that FOXO1 is essential for the maintenance of human ESC pluripotency, and that an orthologue of FOXO1 (Foxo1) exerts a similar function in mouse ESCs. This function is probably mediated through direct control by FOXO1 of OCT4 and SOX2 gene expression through occupation and activation of their respective promoters. Finally, AKT is not the predominant regulator of FOXO1 in human ESCs. Together these results indicate that FOXO1 is a component of the circuitry of human ESC pluripotency. These findings have critical implications for stem cell biology, development, longevity and reprogramming, with potentially important ramifications for therapy.

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Acknowledgements

We thank F. Lohmann for advice on the ChIP assay, J. Bieker (Mount Sinai School of Medicine) and G. Blobel (University of Pennsylvania) for critical reading of the manuscript, I. George and M. Grisotto for cell sorting, and the Flow Cytometry Shared Research Facility of Mount Sinai School of Medicine. This work was supported in part by a National Institutes of Health grant RO1 DK077174, an American Cancer Society Research Scholarship (RSG LIB-110480), a Career Enhancement Award (K18 HL76510-01), a Black Family Stem Cell Institute Exploratory Research Award, a New York State Stem Cell Science (NYSTEM) award (CO24408), an Irma Hirschl/Weill-Caulier Trust Research Award and a Roche Foundation for Anemia Research (RoFAR) Award to S.G., and an NIH P20 GM75019 (S.G. CoPI).

Author information

Author notes

    • Markus Landthaler

    Present address: Berlin Institute for Medical Systems Biology at the Max-Delbrück-Center for Molecular Medicine, 13092 Berlin, Germany

Affiliations

  1. Department of Developmental and Regenerative Biology, Mount Sinai School of Medicine, New York, New York 10029, USA

    • Xin Zhang
    • , Safak Yalcin
    • , Dung-Fang Lee
    • , Seung-Min Lee
    • , Jie Su
    • , Sathish Kumar Mungamuri
    • , Pauline Rimmelé
    • , Ihor Lemischka
    •  & Saghi Ghaffari
  2. Research Service, VA San Diego Healthcare System, School of Medicine, University of California, San Diego, La Jolla, California 92093, USA

    • Tsung-Yin J. Yeh
    •  & Nai-Wen Chi
  3. McEwen Center for Regenerative Medicine, University Health Network, Toronto, Ontario M5G 1L7, Canada

    • Marion Kennedy
    •  & Gordon Keller
  4. Department of Pathology, Albert Einstein College of Medicine, New York, New York 10461, USA

    • Rani Sellers
  5. Howard Hughes Medical Institute, Laboratory for RNA Molecular Biology, The Rockefeller University, New York, New York 10065, USA

    • Markus Landthaler
    •  & Thomas Tuschl
  6. Black Family Stem Cell Institute, Mount Sinai School of Medicine, New York, New York 10029, USA

    • Ihor Lemischka
    •  & Saghi Ghaffari
  7. Department of Medicine Division of Hematology, Oncology, Mount Sinai School of Medicine, New York, New York 10029, USA

    • Saghi Ghaffari

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Contributions

X.Z. and S.G. designed experiments and analysed data; X.Z. carried out most of the experiments, with significant help from S.Y. and some assistance from S-M.L., S.K.M. and P.R.; M.K. helped with the set-up of some critical techniques; R.S. analysed data; D-F.L. and J.S. designed and carried out experiments involving mESCs; N-W.C. designed and carried out antibody calibration experiments with the help of T-Y.J.Y.; M.L. and T.T. contributed key reagents; I.L. and G.K. provided valuable reagents and advice; S.G. conceived the project and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Saghi Ghaffari.

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DOI

https://doi.org/10.1038/ncb2293

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