Abstract

Stem-cell fate can be influenced by metabolite levels in culture, but it is not known whether physiological variations in metabolite levels in normal tissues regulate stem-cell function in vivo. Here we describe a metabolomics method for the analysis of rare cell populations isolated directly from tissues and use it to compare mouse haematopoietic stem cells (HSCs) to restricted haematopoietic progenitors. Each haematopoietic cell type had a distinct metabolic signature. Human and mouse HSCs had unusually high levels of ascorbate, which decreased with differentiation. Systemic ascorbate depletion in mice increased HSC frequency and function, in part by reducing the function of Tet2, a dioxygenase tumour suppressor. Ascorbate depletion cooperated with Flt3 internal tandem duplication (Flt3ITD) leukaemic mutations to accelerate leukaemogenesis, through cell-autonomous and possibly non-cell-autonomous mechanisms, in a manner that was reversed by dietary ascorbate. Ascorbate acted cell-autonomously to negatively regulate HSC function and myelopoiesis through Tet2-dependent and Tet2-independent mechanisms. Ascorbate therefore accumulates within HSCs to promote Tet activity in vivo, limiting HSC frequency and suppressing leukaemogenesis.

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Acknowledgements

S.J.M. is a Howard Hughes Medical Institute (HHMI) Investigator, the Mary McDermott Cook Chair in Pediatric Genetics, the Kathryn and Gene Bishop Distinguished Chair in Pediatric Research, the director of the Hamon Laboratory for Stem Cells and Cancer, and a Cancer Prevention and Research Institute of Texas Scholar. M.A. was a Royal Commission for the Exhibition of 1851 Research Fellow. We thank F. Harrison for sharing the Slc23a2/ mice, N. Loof and the Moody Foundation Flow Cytometry Facility for flow cytometry, K. Correll, A. Leach and A. Gross for mouse colony management and BioHPC at UT Southwestern for providing high-performance computing. This work was supported by the Cancer Prevention and Research Institute of Texas and the National Institutes of Health (R37 AG024945 and R01 DK100848).

Author information

Affiliations

  1. Children’s Research Institute and the Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA

    • Michalis Agathocleous
    • , Corbin E. Meacham
    • , Rebecca J. Burgess
    • , Elena Piskounova
    • , Zhiyu Zhao
    • , Genevieve M. Crane
    • , Brianna L. Cowin
    • , Emily Bruner
    • , Malea M. Murphy
    • , Zeping Hu
    • , Ralph J. DeBerardinis
    •  & Sean J. Morrison
  2. Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA

    • Weina Chen
  3. Department of Medicine, University of Utah, Salt Lake City, Utah, USA

    • Gerald J. Spangrude
  4. Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA

    • Ralph J. DeBerardinis
    •  & Sean J. Morrison

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Contributions

M.A. conceived and performed most experiments. C.E.M. performed experiments with Tet2fl and Tet2fl;Flt3ITD mice in Extended Data Figs 5, 8. R.J.B. and E.P. performed the histone methylation analysis. E.P. and Z.Z. performed RNA-sequencing analysis. Z.Z. performed the statistical analyses. G.M.C. assessed haematopathology in Figs 4, 5. E.B. and B.L.C. provided technical assistance. M.M.M. performed collagen staining. W.C. provided human bone marrow specimens. G.J.S. provided some of the data from Slc23a2/ mice. Z.H., M.A., and R.J.D. developed the metabolomics methods and R.J.D. helped to interpret metabolomics results. M.A. and S.J.M. designed experiments, interpreted results and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Sean J. Morrison.

Reviewer Information Nature thanks H. Christofk, R. Levine and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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Extended data

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    Supplementary Information

    This file contains the uncropped gels, representative flow cytometry plots and a list of antibodies used.

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https://doi.org/10.1038/nature23876

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