Abstract

D-type cyclins (D1, D2 and D3) and their associated cyclin-dependent kinases (CDK4 and CDK6) are components of the core cell cycle machinery that drives cell proliferation1,2. Inhibitors of CDK4 and CDK6 are currently being tested in clinical trials for patients with several cancer types, with promising results2. Here, using human cancer cells and patient-derived xenografts in mice, we show that the cyclin D3–CDK6 kinase phosphorylates and inhibits the catalytic activity of two key enzymes in the glycolytic pathway, 6-phosphofructokinase and pyruvate kinase M2. This re-directs the glycolytic intermediates into the pentose phosphate (PPP) and serine pathways. Inhibition of cyclin D3–CDK6 in tumour cells reduces flow through the PPP and serine pathways, thereby depleting the antioxidants NADPH and glutathione. This, in turn, increases the levels of reactive oxygen species and causes apoptosis of tumour cells. The pro-survival function of cyclin D-associated kinase operates in tumours expressing high levels of cyclin D3–CDK6 complexes. We propose that measuring the levels of cyclin D3–CDK6 in human cancers might help to identify tumour subsets that undergo cell death and tumour regression upon inhibition of CDK4 and CDK6. Cyclin D3–CDK6, through its ability to link cell cycle and cell metabolism, represents a particularly powerful oncoprotein that affects cancer cells at several levels, and this property can be exploited for anti-cancer therapy.

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Acknowledgements

Supported by R01 CA083688, R01 CA202634, P01 CA080111 (P.S.), R01 CA163698 (N.J.D.) and F32 CA165856 (B.N.N.). N.J.D. is a James and Shirley Curvey MGH Research Scholar. X.G. was supported by an NIH post-doc training grant (T32CA009361) and NIH grant P50 CA090381-14 (DF/HCC SPORE in Prostate Cancer). J.M.S. was supported by a Mobilnos´c´ Plus fellowship. We thank M. Eck, J. Daly, P. Hydbring, T. Otto, W. Michowski and I. Harris for help.

Author information

Affiliations

  1. Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA

    • Haizhen Wang
    • , Xueliang Gao
    • , Yan Geng
    • , Hong Ren
    • , Jan M. Suski
    • , Thomas M. Roberts
    •  & Piotr Sicinski
  2. Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Haizhen Wang
    • , Yan Geng
    • , Hong Ren
    • , Jan M. Suski
    •  & Piotr Sicinski
  3. Massachusetts General Hospital Cancer Center, Charlestown, Massachusetts 02129, USA

    • Brandon N. Nicolay
    •  & Nicholas J. Dyson
  4. Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Joel M. Chick
    •  & Steven P. Gygi
  5. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Xueliang Gao
    •  & Thomas M. Roberts
  6. Novartis Institutes for Biomedical Research, Cambridge, Massachusetts 02139, USA

    • Hui Gao
    • , Guizhi Yang
    •  & Juliet A. Williams
  7. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 USA

    • Mark A. Keibler
  8. Department of Oncologic Pathology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA

    • Ewa Sicinska
  9. Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA

    • Ulrike Gerdemann
    •  & W. Nicholas Haining
  10. Division of Pediatric Hematology and Oncology, Children’s Hospital, Boston, Massachusetts 02115, USA

    • W. Nicholas Haining
  11. Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, USA

    • W. Nicholas Haining
  12. Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA

    • Kornelia Polyak

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Contributions

H.W. and P.S. designed the study. H.W. performed all experiments with the help of collaborators as follows. B.N.N. and N.J.D. performed and interpreted isotopic enrichment analyses. J.M.C. and S.P.G. contributed mass spectrometric and biocomputational analyses. X.G. helped with viral transductions and kinase assays. Y.G. helped with T-ALL xenografts and kinase assays. H.R. helped with analyses of cyclin levels and tissue culture. J.M.S. helped with design and construction of expression vectors. T.M.R. helped with supervision. H.G., G.Y. and J.A.W. contributed ribociclib xenograft studies. M.A.K. contributed some isotopic-enrichment analyses. E.S. carried out pathological analyses. U.G. and W.N.H. isolated human T cells. K.P. helped with breast cancer studies. H.W. and P.S. wrote the paper. P.S. directed the study.

Competing interests

P.S., T.M.R. and K.P. are consultants and recipients of research grants from Novartis. H.G., G.Y. and J.A.W. are employees of Novartis Institutes for BioMedical Research.

Corresponding author

Correspondence to Piotr Sicinski.

Reviewer Information Nature thanks J. Bartek, H. Christofk and A. Schulze for their contribution to the peer review of this work.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Figure

    This file contains the uncropped blots.

Excel files

  1. 1.

    Supplementary Table 1

    This tablecontains affinity purification tandem mass spectrometry analysis of CDK6 interactome in T-ALL cells.

  2. 2.

    Supplementary Table 2

    This table contains gene set enrichment analysis of common CDK6 interactors.

  3. 3.

    Supplementary Table 3

    This table contains CDK6-associated glycolytic enzymes that contain potential CDK phosphorylation residues.

  4. 4.

    Supplementary Table 4

    This table contains LC-MS/MS analysis of cyclin D3-CDK6 dependent phosphorylation sites on PKM2 and PFKP.

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DOI

https://doi.org/10.1038/nature22797

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