A key role for mitochondrial gatekeeper pyruvate dehydrogenase in oncogene-induced senescence

Subjects

Abstract

In response to tenacious stress signals, such as the unscheduled activation of oncogenes, cells can mobilize tumour suppressor networks to avert the hazard of malignant transformation. A large body of evidence indicates that oncogene-induced senescence (OIS) acts as such a break, withdrawing cells from the proliferative pool almost irreversibly, thus crafting a vital pathophysiological mechanism that protects against cancer1,2,3,4,5. Despite the widespread contribution of OIS to the cessation of tumorigenic expansion in animal models and humans, we have only just begun to define the underlying mechanism and identify key players6. Although deregulation of metabolism is intimately linked to the proliferative capacity of cells7,8,9,10, and senescent cells are thought to remain metabolically active11, little has been investigated in detail about the role of cellular metabolism in OIS. Here we show, by metabolic profiling and functional perturbations, that the mitochondrial gatekeeper pyruvate dehydrogenase (PDH) is a crucial mediator of senescence induced by BRAFV600E, an oncogene commonly mutated in melanoma and other cancers. BRAFV600E-induced senescence was accompanied by simultaneous suppression of the PDH-inhibitory enzyme pyruvate dehydrogenase kinase 1 (PDK1) and induction of the PDH-activating enzyme pyruvate dehydrogenase phosphatase 2 (PDP2). The resulting combined activation of PDH enhanced the use of pyruvate in the tricarboxylic acid cycle, causing increased respiration and redox stress. Abrogation of OIS, a rate-limiting step towards oncogenic transformation, coincided with reversion of these processes. Further supporting a crucial role of PDH in OIS, enforced normalization of either PDK1 or PDP2 expression levels inhibited PDH and abrogated OIS, thereby licensing BRAFV600E-driven melanoma development. Finally, depletion of PDK1 eradicated melanoma subpopulations resistant to targeted BRAF inhibition, and caused regression of established melanomas. These results reveal a mechanistic relationship between OIS and a key metabolic signalling axis, which may be exploited therapeutically.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: The PDK1–PDP2–PDH axis is deregulated in OIS.
Figure 2: PDP2 regulates metabolic rewiring and OIS.
Figure 3: PDK1 regulates metabolic rewiring and OIS, and acts tumorigenically.
Figure 4: PDK1 depletion causes melanoma regression and eradicates subpopulations resistant to targeted BRAFV600E inhibition.

References

  1. 1

    Campisi, J. Suppressing cancer: the importance of being senescent. Science 309, 886–887 (2005)

    CAS  Article  Google Scholar 

  2. 2

    Collado, M. & Serrano, M. Senescence in tumours: evidence from mice and humans. Nature Rev. Cancer 10, 51–57 (2010)

    CAS  Article  Google Scholar 

  3. 3

    Vredeveld, L. C. W. et al. Abrogation of BRAFV600E-induced senescence by PI3K pathway activation contributes to melanomagenesis. Genes Dev. 26, 1055–1069 (2012)

    CAS  Article  Google Scholar 

  4. 4

    Kuilman, T., Michaloglou, C., Mooi, W. J. & Peeper, D. S. The essence of senescence. Genes Dev. 24, 2463–2479 (2010)

    CAS  Article  Google Scholar 

  5. 5

    Lowe, S. W., Cepero, E. & Evan, G. Intrinsic tumour suppression. Nature 432, 307–315 (2004)

    ADS  CAS  Article  Google Scholar 

  6. 6

    Adams, P. D. Healing and hurting: molecular mechanisms, functions, and pathologies of cellular senescence. Mol. Cell 36, 2–14 (2009)

    CAS  Article  Google Scholar 

  7. 7

    DeBerardinis, R. J., Sayed, N., Ditsworth, D. & Thompson, C. B. Brick by brick: metabolism and tumor cell growth. Curr. Opin. Genet. Dev. 18, 54–61 (2008)

    CAS  Article  Google Scholar 

  8. 8

    Tennant, D. A., Durán, R. V. & Gottlieb, E. Targeting metabolic transformation for cancer therapy. Nature Rev. Cancer 10, 267–277 (2010)

    CAS  Article  Google Scholar 

  9. 9

    Wellen, K. E. & Thompson, C. B. Cellular metabolic stress: considering how cells respond to nutrient excess. Mol. Cell 40, 323–332 (2010)

    CAS  Article  Google Scholar 

  10. 10

    Vander Heiden, M. G., Cantley, L. C. & Thompson, C. B. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324, 1029–1033 (2009)

    ADS  CAS  Article  Google Scholar 

  11. 11

    Campisi, J. Replicative senescence: an old lives’ tale? Cell 84, 497–500 (1996)

    CAS  Article  Google Scholar 

  12. 12

    Michaloglou, C. et al. BRAFE600-associated senescence-like cell cycle arrest of human naevi. Nature 436, 720–724 (2005)

    ADS  CAS  Article  Google Scholar 

  13. 13

    Kuilman, T. et al. Oncogene-induced senescence relayed by an interleukin-dependent inflammatory network. Cell 133, 1019–1031 (2008)

    CAS  Article  Google Scholar 

  14. 14

    Dankort, D. et al. BrafV600E cooperates with Pten loss to induce metastatic melanoma. Nature Genet. 41, 544–552 (2009)

    CAS  Article  Google Scholar 

  15. 15

    Dhomen, N. et al. Oncogenic Braf induces melanocyte senescence and melanoma in mice. Cancer Cell 15, 294–303 (2009)

    CAS  Article  Google Scholar 

  16. 16

    Davies, H. et al. Mutations of the BRAF gene in human cancer. Nature 417, 949–954 (2002)

    ADS  CAS  Article  Google Scholar 

  17. 17

    Wieland, O. H. The mammalian pyruvate dehydrogenase complex: structure and regulation. Rev. Physiol. Biochem. Pharmacol. 96, 123–170 (1983)

    CAS  Article  Google Scholar 

  18. 18

    Patel, M. S. & Roche, T. E. Molecular biology and biochemistry of pyruvate dehydrogenase complexes. FASEB J. 4, 3224–3233 (1990)

    CAS  Article  Google Scholar 

  19. 19

    Kolobova, E., Tuganova, A., Boulatnikov, I. & Popov, K. M. Regulation of pyruvate dehydrogenase activity through phosphorylation at multiple sites. Biochem. J. 358, 69–77 (2001)

    CAS  Article  Google Scholar 

  20. 20

    Roche, T. E. et al. Distinct regulatory properties of pyruvate dehydrogenase kinase and phosphatase isoforms. Prog. Nucleic Acid Res. Mol. Biol. 70, 33–75 (2001)

    CAS  Article  Google Scholar 

  21. 21

    Holness, M. J. & Sugden, M. C. Regulation of pyruvate dehydrogenase complex activity by reversible phosphorylation. Biochem. Soc. Trans. 31, 1143–1151 (2003)

    CAS  Article  Google Scholar 

  22. 22

    Patel, M. S. & Korotchkina, L. G. Regulation of the pyruvate dehydrogenase complex. Biochem. Soc. Trans. 34, 217–222 (2006)

    CAS  Article  Google Scholar 

  23. 23

    Lemons, J. M. S. et al. Quiescent fibroblasts exhibit high metabolic activity. PLoS Biol. 8, e1000514 (2010)

    Article  Google Scholar 

  24. 24

    Fantin, V. R., St-Pierre, J. & Leder, P. Attenuation of LDH-A expression uncovers a link between glycolysis, mitochondrial physiology, and tumor maintenance. Cancer Cell 9, 425–434 (2006)

    CAS  Article  Google Scholar 

  25. 25

    Mooi, W. J. & Peeper, D. S. Oncogene-induced cell senescence–halting on the road to cancer. N. Engl. J. Med. 355, 1037–1046 (2006)

    CAS  Article  Google Scholar 

  26. 26

    Tsai, J. et al. Discovery of a selective inhibitor of oncogenic B-Raf kinase with potent antimelanoma activity. Proc. Natl Acad. Sci. USA 105, 3041–3046 (2008)

    ADS  CAS  Article  Google Scholar 

  27. 27

    Flaherty, K. T., Yasothan, U. & Kirkpatrick, P. Vemurafenib. Nature Rev. Drug Discov. 10, 811–812 (2011)

    CAS  Article  Google Scholar 

  28. 28

    Frezza, C. et al. Haem oxygenase is synthetically lethal with the tumour suppressor fumarate hydratase. Nature 477, 225–228 (2011)

    ADS  CAS  Article  Google Scholar 

  29. 29

    Frezza, C. et al. Metabolic profiling of hypoxic cells revealed a catabolic signature required for cell survival. PLoS ONE 6, e24411 (2011)

    ADS  CAS  Article  Google Scholar 

  30. 30

    Chaneton, B. et al. Serine is a natural ligand and allosteric activator of pyruvate kinase M2. Nature 491, 458–462 (2012)

    ADS  CAS  Article  Google Scholar 

  31. 31

    Serrano, M., Lin, A. W., McCurrach, M. E., Beach, D. & Lowe, S. W. Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88, 593–602 (1997)

    CAS  Article  Google Scholar 

  32. 32

    Sviderskaya, E. V. et al. Complementation of hypopigmentation in p-mutant (pink-eyed dilution) mouse melanocytes by normal human P cDNA, and defective complementation by OCA2 mutant sequences. J. Invest. Dermatol. 108, 30–34 (1997)

    CAS  Article  Google Scholar 

  33. 33

    Dankort, D. et al. A new mouse model to explore the initiation, progression, and therapy of BRAFV600E-induced lung tumors. Genes Dev. 21, 379–384 (2007)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank J.-Y. Song for pathological analysis, M. McMahon for providing BrafCA mice, C. Vogel for sharing cell lines, R. van Amerongen for critical reading of the manuscript, and all members of the Gottlieb and Peeper laboratories for their input. This work was supported by Cancer Research UK, Spanish Government-EU-FEDER (grants SAF2011-25726 and ISCIII-RTICC-RD6/0020/0046) and ICREA-Academia to M.C., Israel Cancer Research Foundation and Israel Science Foundation to T.S., a Vici grant from the Netherlands Organization for Scientific Research (NWO) and a Queen Wilhelmina Award grant from the Dutch Cancer Society (KWF Kankerbestrijding) to D.S.P.

Author information

Affiliations

Authors

Contributions

J.K., E.G. and D.S.P. conceived the project, analysed the data and wrote the manuscript. J.K. performed all in vitro experiments and carried out the in vivo experiments together with K.M. J.K., K.M. and B.C. performed metabolic experiments. L.Z. and G.M. performed LC–MS analyses. S.H.B. and E.M.E.V. provided low passage melanoma cell lines. V.A.S., M.C. and T.S. helped with metabolic analyses. All authors discussed the results and commented on the manuscript. E.G. and D.S.P. contributed equally to this work.

Corresponding authors

Correspondence to Eyal Gottlieb or Daniel S. Peeper.

Ethics declarations

Competing interests

A patent application for combined PDK and MAPK/ERK pathway inhibition in neoplasia has been filed, with J.K. and D.S.P. as inventors.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1-15. (PDF 4632 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kaplon, J., Zheng, L., Meissl, K. et al. A key role for mitochondrial gatekeeper pyruvate dehydrogenase in oncogene-induced senescence. Nature 498, 109–112 (2013). https://doi.org/10.1038/nature12154

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.