Abstract

Most differentiated cells convert glucose to pyruvate in the cytosol through glycolysis, followed by pyruvate oxidation in the mitochondria. These processes are linked by the mitochondrial pyruvate carrier (MPC), which is required for efficient mitochondrial pyruvate uptake. In contrast, proliferative cells, including many cancer and stem cells, perform glycolysis robustly but limit fractional mitochondrial pyruvate oxidation. We sought to understand the role this transition from glycolysis to pyruvate oxidation plays in stem cell maintenance and differentiation. Loss of the MPC in Lgr5-EGFP-positive stem cells, or treatment of intestinal organoids with an MPC inhibitor, increases proliferation and expands the stem cell compartment. Similarly, genetic deletion of the MPC in Drosophila intestinal stem cells also increases proliferation, whereas MPC overexpression suppresses stem cell proliferation. These data demonstrate that limiting mitochondrial pyruvate metabolism is necessary and sufficient to maintain the proliferation of intestinal stem cells.

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Acknowledgements

We thank B. Edgar (University of Utah, USA) for stocks and reagents, C. Micchelli (Washington University School of Medicine, USA) for providing the Notch RNAi line, K. Beebe for helpful advice and comments on the Drosophila intestinal studies, G. Lam for establishing the Drosophila MPC overexpression strain, O. Yilmaz and D. Sabatini for assistance and insight into intestinal stem cell metabolism, D. Tantin for critiques and comments, members of the Rutter laboratory for assistance and advice, J. O’Shea, R. Orbus and C. DeHeer for assistance with NanoString, W. Swiatek for mouse assistance, ARUP Institute for Clinical and Experimental Pathology, and S. R. Tripp and E. Hammond for histology; L. Nikolova at the University of Utah Electron Microscopy Core Laboratory performed electron microscopy; mass spectrometry analysis was performed at the Mass Spectrometry and Proteomics Core Facility at the University of Utah. Mass spectrometry equipment was obtained through NCRR Shared Instrumentation Grant no. 1 S10 RR020883-01, 1 S10 RR025532-01A1, NIH 1 S10OD021505-01 (J.E.C.) and the Diabetes and Metabolism Center at the University of Utah. This study was conducted with support from the Biorepository and Molecular Pathology Shared Resource supported by the Cancer Center Support Grant awarded to the Huntsman Cancer Institute by the National Cancer Institute of the National Institutes of Health. Nanostring transcript analysis utilized the Molecular Diagnostics Section of the Biorepository and Molecular Pathology Shared Resource and was supported by the National Cancer Institute of the National Institutes of Health under Award Number P30CA042014 (the content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH). J. Marvin at the University of Utah Flow Cytometry Facility carried out flow sorting (National Cancer Institute through Award Number 5P30CA042014-24, National Center for Research Resources of the National Institutes of Health under Award Number 1S10RR026802-01). Funding was also provided by HHMI (J.R.), Treadwell (J.R.) and RO1GM094232 (to J.R. and C.S.T.). J.C.S. was supported by an NIH Developmental Biology Training Grant (5T32 HD07491) and a University of Utah Graduate Research Fellowship. D.R.W. was supported by a University of Utah Graduate Research Fellowship.

Author information

Affiliations

  1. Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah 84112, USA

    • John C. Schell
    • , Claire Bensard
    • , Peng Wei
    • , Jason Tanner
    • , Christian S. Earl
    • , Kristofor A. Olson
    • , Ren Miao
    • , T. Cameron Waller
    • , James E. Cox
    •  & Jared Rutter
  2. Department of Human Genetics, University of Utah School of Medicine, Salt Lake City, Utah 84112, USA

    • Dona R. Wisidagama
    •  & Carl S. Thummel
  3. Department of Molecular Medicine, University of Utah School of Medicine, Salt Lake City, Utah 84112, USA

    • Helong Zhao
    • , Lise K. Sorensen
    •  & Dean Y. Li
  4. Eli and Edythe Broad Center for Regenerative Medicine, University of California Los Angeles, Los Angeles, California 90095, USA

    • Aimee Flores
    • , Heather R. Christofk
    •  & William E. Lowry
  5. Department of Molecular Cell and Developmental Biology, University of California Los Angeles, Los Angeles, California 90095, USA

    • Aimee Flores
    •  & William E. Lowry
  6. Department of Pathology, University of Utah School of Medicine, Salt Lake City, Utah 84112, USA

    • Jeffrey Mohlman
    •  & Mary P. Bronner
  7. Department of Internal Medicine, University of Utah School of Medicine, Salt Lake City, Utah 84112, USA

    • Don Delker
    •  & Priyanka Kanth
  8. Children’s Medical Center Research Institute, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA

    • Lei Jiang
    •  & Ralph J. DeBerardinis
  9. Department of Molecular and Cellular Endocrinology, Beckman Research Institute at City of Hope, Duarte, California 91010, USA

    • Lei Jiang
  10. Department of Biological Chemistry, University of California Los Angeles, Los Angeles, California 90095, USA

    • Heather R. Christofk
  11. Howard Hughes Medical Institute, University of Utah School of Medicine, Salt Lake City, Utah 84112, USA

    • Jared Rutter

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Contributions

Conceptualization, J.C.S., D.R.W., C.S.T. and J.R.; methodology, J.C.S., D.R.W., C.B., H.Z., P.W., J.T., A.F., J.M., L.K.S., C.S.E., K.A.O., D.D., P.K., M.P.B., D.Y.L., J.E.C., H.R.C., W.E.L., C.S.T. and J.R.; investigation, J.C.S., D.R.W., C.B., H.Z., P.W., J.T., A.F., J.M., L.K.S., C.S.E., R.M., D.D. and P.K.; formal analysis, J.C.S., D.R.W., C.B., P.W., T.C.W., R.M., L.J., R.J.D. and J.E.C.; writing—original draft, review and editing, J.C.S., D.R.W., C.S.T. and J.R.; funding acquisition, C.S.T. and J.R.; resources, D.Y.L., J.C., C.S.T. and J.R.; supervision, C.S.T. and J.R.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Jared Rutter.

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https://doi.org/10.1038/ncb3593

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