Letter | Published:

Metabolic determinants of cancer cell sensitivity to glucose limitation and biguanides

Nature volume 508, pages 108112 (03 April 2014) | Download Citation

Abstract

As the concentrations of highly consumed nutrients, particularly glucose, are generally lower in tumours than in normal tissues1,2, cancer cells must adapt their metabolism to the tumour microenvironment. A better understanding of these adaptations might reveal cancer cell liabilities that can be exploited for therapeutic benefit. Here we developed a continuous-flow culture apparatus (Nutrostat) for maintaining proliferating cells in low-nutrient media for long periods of time, and used it to undertake competitive proliferation assays on a pooled collection of barcoded cancer cell lines cultured in low-glucose conditions. Sensitivity to low glucose varies amongst cell lines, and an RNA interference (RNAi) screen pinpointed mitochondrial oxidative phosphorylation (OXPHOS) as the major pathway required for optimal proliferation in low glucose. We found that cell lines most sensitive to low glucose are defective in the OXPHOS upregulation that is normally caused by glucose limitation as a result of either mitochondrial DNA (mtDNA) mutations in complex I genes or impaired glucose utilization. These defects predict sensitivity to biguanides, antidiabetic drugs that inhibit OXPHOS3,4, when cancer cells are grown in low glucose or as tumour xenografts. Notably, the biguanide sensitivity of cancer cells with mtDNA mutations was reversed by ectopic expression of yeast NDI1, a ubiquinone oxidoreductase that allows bypass of complex I function5. Thus, we conclude that mtDNA mutations and impaired glucose utilization are potential biomarkers for identifying tumours with increased sensitivity to OXPHOS inhibitors.

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Acknowledgements

We thank G. Stephanopoulos for assistance with Nutrostat design, M. Holland for mtDNA sequencing consultation, L. Garraway for assistance identifying mtDNA mutant cell lines, T. Yagi for the NDI1 antibody, T. DiCesare for diagrams, and members of the Sabatini Laboratory for assistance (particularly A. Saucedo, C. Koch, O. Yilmaz, Y. Gultekin and A. Hutchins for technical assistance, and D. Lamming and W. Comb for critical reading of the manuscript). This research is supported by fellowships from The Leukemia and Lymphoma Society and The Jane Coffin Childs Fund to K.B., the Council of Higher Education Turkey and Karadeniz T. University Scholarships to B.Y. and grants from the David H. Koch Institute for Integrative Cancer Research at MIT, The Alexander and Margaret Stewart Trust Fund, the NIH (K99 CA168940 to R.P. and CA103866, CA129105, and AI07389 to D.M.S.) and the Starr Cancer Consortium. D.M.S. is an investigator of the Howard Hughes Medical Institute.

Author information

Author notes

    • Kıvanç Birsoy
    •  & Richard Possemato

    These authors contributed equally to this work.

Affiliations

  1. Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA

    • Kıvanç Birsoy
    • , Richard Possemato
    • , Franziska K. Lorbeer
    • , Erol C. Bayraktar
    • , Prathapan Thiru
    • , Burcu Yucel
    • , Tim Wang
    • , Walter W. Chen
    •  & David M. Sabatini
  2. Howard Hughes Medical Institute and Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA

    • Kıvanç Birsoy
    • , Richard Possemato
    • , Tim Wang
    • , Walter W. Chen
    •  & David M. Sabatini
  3. Broad Institute of Harvard and MIT, Seven Cambridge Center, Cambridge, Massachusetts 02142, USA

    • Kıvanç Birsoy
    • , Richard Possemato
    • , Tim Wang
    • , Walter W. Chen
    • , Clary B. Clish
    •  & David M. Sabatini
  4. The David H. Koch Institute for Integrative Cancer Research at MIT, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA

    • Kıvanç Birsoy
    • , Richard Possemato
    • , Tim Wang
    • , Walter W. Chen
    •  & David M. Sabatini

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Contributions

K.B., R.P. and D.M.S. conceived the project and designed the experiments. K.B. and R.P. designed and engineered the Nutrostat and performed the screening, knockdown, cell proliferation, extracellular flux, glucose consumption, and tumour formation experiments and processed and analysed sequencing and expression data. F.K.L., E.C.B., B.Y., and W.W.C. assisted with experiments. C.B.C. performed the metabolite profiling experiments. T.W. provided bioinformatic support for shRNA abundance deconvolution, P.T. assisted in identifying mtDNA mutations. K.B., R.L.P and D.M.S. wrote and all authors edited the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to David M. Sabatini.

Extended data

Supplementary information

Excel files

  1. 1.

    Supplementary Table 1

    Primary Cell Competition Data. Primary Data for Figure 2b, Proliferation of barcoded cell lines in high and low glucose.

  2. 2.

    Supplementary Table 2

    Primary Screening Data. Primary Data for Figure 2d, RNAi Screen under 10 mM or 0.75 mM glucose conditions.

  3. 3.

    Supplementary Table 3

    Alternative Hit Lists from Pooled shRNA Screen. Data from Supplementary Table 2 analyzed using the GENE-E software to generate lists of scoring genes alternative to those in Figure 2e and Extended Data Figure 2a.

  4. 4.

    Supplementary Table 4

    Impaired Glucose Import Gene Signature Data. Primary Data for Extended Data Figure 5.

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DOI

https://doi.org/10.1038/nature13110

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