Pyruvate kinase M2 activators promote tetramer formation and suppress tumorigenesis (original) (raw)

Accession codes

Primary accessions

Protein Data Bank

Referenced accessions

Protein Data Bank

Change history

In the version of this article initially published, the accession codes to the Protein Data Bank were omitted. The error has been corrected in the HTML and PDF versions of the article.

References

  1. Tennant, D.A., Duran, R.V. & Gottlieb, E. Targeting metabolic transformation for cancer therapy. Nat. Rev. Cancer 10, 267–277 (2010).
    Article CAS PubMed Google Scholar
  2. Vander Heiden, M.G. Targeting cancer metabolism: a therapeutic window opens. Nat. Rev. Drug Discov. 10, 671–684 (2011).
    Article CAS PubMed Google Scholar
  3. 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).
    CAS PubMed PubMed Central Google Scholar
  4. Cairns, R.A., Harris, I.S. & Mak, T.W. Regulation of cancer cell metabolism. Nat. Rev. Cancer 11, 85–95 (2011).
    Article CAS PubMed Google Scholar
  5. Levine, A.J. & Puzio-Kuter, A.M. The control of the metabolic switch in cancers by oncogenes and tumor suppressor genes. Science 330, 1340–1344 (2010).
    Article CAS PubMed Google Scholar
  6. Trachootham, D., Alexandre, J. & Huang, P. Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach? Nat. Rev. Drug Discov. 8, 579–591 (2009).
    Article CAS PubMed Google Scholar
  7. Weissleder, R. Molecular imaging in cancer. Science 312, 1168–1171 (2006).
    Article CAS PubMed Google Scholar
  8. Christofk, H.R. et al. The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth. Nature 452, 230–233 (2008).
    Article CAS PubMed Google Scholar
  9. Mazurek, S. Pyruvate kinase type M2: a key regulator of the metabolic budget system in tumor cells. Int. J. Biochem. Cell Biol. 43, 969–980 (2011).
    Article CAS PubMed Google Scholar
  10. Noguchi, T., Inoue, H. & Tanaka, T. The M1- and M2-type isozymes of rat pyruvate kinase are produced from the same gene by alternative RNA splicing. J. Biol. Chem. 261, 13807–13812 (1986).
    CAS PubMed Google Scholar
  11. Clower, C.V. et al. The alternative splicing repressors hnRNP A1/A2 and PTB influence pyruvate kinase isoform expression and cell metabolism. Proc. Natl. Acad. Sci. USA 107, 1894–1899 (2010).
    Article CAS PubMed PubMed Central Google Scholar
  12. Yamada, K. & Noguchi, T. Regulation of pyruvate kinase M gene expression. Biochem. Biophys. Res. Commun. 256, 257–262 (1999).
    Article CAS PubMed Google Scholar
  13. Ikeda, Y., Tanaka, T. & Noguchi, T. Conversion of non-allosteric pyruvate kinase isozyme into an allosteric enzyme by a single amino acid substitution. J. Biol. Chem. 272, 20495–20501 (1997).
    Article CAS PubMed Google Scholar
  14. Ikeda, Y. & Noguchi, T. Allosteric regulation of pyruvate kinase M2 isozyme involves a cysteine residue in the intersubunit contact. J. Biol. Chem. 273, 12227–12233 (1998).
    Article CAS PubMed Google Scholar
  15. Ashizawa, K., Willingham, M.C., Liang, C.M. & Cheng, S.Y. In vivo regulation of monomer-tetramer conversion of pyruvate kinase subtype M2 by glucose is mediated via fructose 1,6-bisphosphate. J. Biol. Chem. 266, 16842–16846 (1991).
    CAS PubMed Google Scholar
  16. Ashizawa, K., McPhie, P., Lin, K.H. & Cheng, S.Y. An in vitro novel mechanism of regulating the activity of pyruvate kinase M2 by thyroid hormone and fructose 1, 6-bisphosphate. Biochemistry 30, 7105–7111 (1991).
    Article CAS PubMed Google Scholar
  17. Christofk, H.R., Vander Heiden, M.G., Wu, N., Asara, J.M. & Cantley, L.C. Pyruvate kinase M2 is a phosphotyrosine-binding protein. Nature 452, 181–186 (2008).
    Article CAS PubMed Google Scholar
  18. Eigenbrodt, E., Reinacher, M., Scheefers-Borchel, U., Scheefers, H. & Friis, R. Double role for pyruvate kinase type M2 in the expansion of phosphometabolite pools found in tumor cells. Crit. Rev. Oncog. 3, 91–115 (1992).
    CAS PubMed Google Scholar
  19. Anastasiou, D. et al. Inhibition of pyruvate kinase M2 by reactive oxygen species contributes to cellular antioxidant responses. Science 334, 1278–1283 (2011).
    Article CAS PubMed PubMed Central Google Scholar
  20. Vander Heiden, M.G. et al. Evidence for an alternative glycolytic pathway in rapidly proliferating cells. Science 329, 1492–1499 (2010).
    Article CAS PubMed Google Scholar
  21. Locasale, J.W., Vander Heiden, M.G. & Cantley, L.C. Rewiring of glycolysis in cancer cell metabolism. Cell Cycle 9, 4253 (2010).
    Article CAS PubMed Google Scholar
  22. Jiang, J.K. et al. Evaluation of thieno[3,2-b]pyrrole[3,2-d]pyridazinones as activators of the tumor cell specific M2 isoform of pyruvate kinase. Bioorg. Med. Chem. Lett. 20, 3387–3393 (2010).
    Article CAS PubMed PubMed Central Google Scholar
  23. Boxer, M.B. et al. Evaluation of substituted N,_N_′-diarylsulfonamides as activators of the tumor cell specific M2 isoform of pyruvate kinase. J. Med. Chem. 53, 1048–1055 (2010).
    Article CAS PubMed PubMed Central Google Scholar
  24. Ikeda, Y., Taniguchi, N. & Noguchi, T. Dominant negative role of the glutamic acid residue conserved in the pyruvate kinase M(1) isozyme in the heterotropic allosteric effect involving fructose-1,6-bisphosphate. J. Biol. Chem. 275, 9150–9156 (2000).
    Article CAS PubMed Google Scholar
  25. Kato, H., Fukuda, T., Parkison, C., McPhie, P. & Cheng, S.Y. Cytosolic thyroid hormone-binding protein is a monomer of pyruvate kinase. Proc. Natl. Acad. Sci. USA 86, 7861–7865 (1989).
    Article CAS PubMed PubMed Central Google Scholar
  26. Hitosugi, T. et al. Tyrosine phosphorylation inhibits PKM2 to promote the Warburg effect and tumor growth. Sci. Signal. 2, ra73 (2009).
    Article PubMed PubMed Central Google Scholar
  27. Lv, L. et al. Acetylation targets the M2 isoform of pyruvate kinase for degradation through chaperone-mediated autophagy and promotes tumor growth. Mol. Cell 42, 719–730 (2011).
    Article CAS PubMed PubMed Central Google Scholar
  28. Luo, W. et al. Pyruvate kinase M2 is a PHD3-stimulated coactivator for hypoxia-inducible factor 1. Cell 145, 732–744 (2011).
    Article CAS PubMed PubMed Central Google Scholar
  29. Hoshino, A., Hirst, J.A. & Fujii, H. Regulation of cell proliferation by interleukin-3–induced nuclear translocation of pyruvate kinase. J. Biol. Chem. 282, 17706–17711 (2007).
    Article CAS PubMed Google Scholar
  30. Steták, A. et al. Nuclear translocation of the tumor marker pyruvate kinase M2 induces programmed cell death. Cancer Res. 67, 1602–1608 (2007).
    Article PubMed Google Scholar
  31. Hatzivassiliou, G. et al. ATP citrate lyase inhibition can suppress tumor cell growth. Cancer Cell 8, 311–321 (2005).
    Article CAS PubMed Google Scholar
  32. Vander Heiden, M.G. et al. Identification of small molecule inhibitors of pyruvate kinase M2. Biochem. Pharmacol. 79, 1118–1124 (2010).
    Article CAS PubMed Google Scholar
  33. Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. in Methods in Enzymology Vol. 276, 307–326 (Academic Press, 1997).
  34. Minor, W., Cymborowski, M., Otwinowski, Z. & Chruszcz, M. HKL-3000: the integration of data reduction and structure solution—from diffraction images to an initial model in minutes. Acta Crystallogr. D Biol. Crystallogr. 62, 859–866 (2006).
    Article PubMed Google Scholar
  35. Berman, H.M. et al. The Protein Data Bank. Nucleic Acids Res. 28, 235–242 (2000).
    Article CAS PubMed PubMed Central Google Scholar
  36. Schüttelkopf, A.W. & van Aalten, D.M. PRODRG: a tool for high-throughput crystallography of protein-ligand complexes. Acta Crystallogr. D Biol. Crystallogr. 60, 1355–1363 (2004).
    Article PubMed Google Scholar
  37. Emsley, P., Lohkamp, B., Scott, W.G. & Cowtan, K. Features and development of Coot. Acta Crystallogr. D Biol. Crystallogr. 66, 486–501 (2010).
    Article CAS PubMed PubMed Central Google Scholar
  38. Murshudov, G.N., Vagin, A.A. & Dodson, E.J. Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr. D Biol. Crystallogr. 53, 240–255 (1997).
    Article CAS PubMed Google Scholar
  39. Davis, I.W., Murray, L.W., Richardson, J.S. & Richardson, D.C. MOLPROBITY: structure validation and all-atom contact analysis for nucleic acids and their complexes. Nucleic Acids Res. 32, W615–W619 (2004).
    Article CAS PubMed PubMed Central Google Scholar
  40. Schumacker, P.T., Chandel, N. & Agusti, A.G. Oxygen conformance of cellular respiration in hepatocytes. Am. J. Physiol. 265, L395–L402 (1993).
    CAS PubMed Google Scholar
  41. Metallo, C.M. et al. Reductive glutamine metabolism by IDH1 mediates lipogenesis under hypoxia. Nature 481, 380–384 (2011).
    Article PubMed PubMed Central Google Scholar
  42. Yuan, M., Breitkopf, S.B., Yang, X. & Asara, J.M. A positive/negative ion-switching, targeted mass spectrometry-based metabolomics platform for bodily fluids, cells, and fresh and fixed tissue. Nat. Protoc. 7, 872–881 (2012).
    Article CAS PubMed PubMed Central Google Scholar
  43. Xia, J., Mandal, R., Sinelnikov, I.V., Broadhurst, D. & Wishart, D.S. MetaboAnalyst 2.0–a comprehensive server for metabolomic data analysis. Nucleic Acids Res. W127–W133 (2012).

Download references

Acknowledgements

The Structural Genomics Consortium is a registered charity (1097737) and receives funds from the Canadian Institutes for Health Research, the Canadian Foundation for Innovation, Genome Canada through the Ontario Genomics Institute, GlaxoSmithKline, Karolinska Institutet, the Knut and Alice Wallenberg Foundation, the Ontario Innovation Trust, the Ontario Ministry for Research and Innovation, Merck and Co., Inc., the Novartis Research Foundation, the Swedish Agency for Innovation Systems, the Swedish Foundation for Strategic Research and the Wellcome Trust. The crystallography results shown in this report are derived from work performed at Argonne National Laboratory, Structural Biology Center at the Advanced Photon Source. Argonne is operated by UChicago Argonne, LLC for the US Department of Energy, Office of Biological and Environmental Research under contract DE-AC02-06CH11357. We thank P. Chang for experimental advice related to sucrose gradient ultracentrifugation and SAI Advantium Pharma Ltd. for help with pharmacokinetics studies. We also thank M. Kini for experimental help and acknowledge M. Yuan and S. Breitkopf for help with MS experiments. This work was supported by the Molecular Libraries Initiative of the NIH Roadmap for Medical Research and the Intramural Research Program of the National Human Genome Research Institute, NIH and by NIH grant R03MH085679. This work was also funded by NIH grant R01 GM56203 (L.C.C.). J.M.A. acknowledges funding from NIH 5P01CA120964 and Dana-Farber/Harvard Cancer Center support grant NIH 5P30CA006516. M.G.V.H. acknowledges additional funding support from the Smith Family Foundation, the Burroughs Wellcome Fund, the Damon Runyon Cancer Research Foundation, the Stern family and the National Cancer Institute, including NIH 5P30CA1405141.

Author information

Author notes

  1. Dimitrios Anastasiou, Yimin Yu and William J Israelsen: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medicine, Division of Signal Transduction, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA
    Dimitrios Anastasiou, Kevin D Courtney, Heather R Christofk, John M Asara & Lewis C Cantley
  2. Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA
    Dimitrios Anastasiou, Kevin D Courtney & Lewis C Cantley
  3. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology Cambridge, Massachusetts, USA
    Yimin Yu, William J Israelsen, Katherine R Mattaini, Brian P Fiske, Scott Malstrom, Tahsin M Khan, Sophia Y Lunt, Zachary R Johnson, Shawn M Davidson & Matthew G Vander Heiden
  4. National Institutes of Health (NIH) Chemical Genomics Center, National Center for Advancing Translational Sciences, NIH, Bethesda, Maryland, USA.,
    Jian-Kang Jiang, Matthew B Boxer, Min Shen, Amanda P Skoumbourdis, Henrike Veith, Noel Southall, Martin J Walsh, Kyle R Brimacombe, William Leister, Christopher P Austin, James Inglese, Douglas S Auld & Craig J Thomas
  5. Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada
    Bum Soo Hong, Wolfram Tempel, Svetoslav Dimov & Hee-Won Park
  6. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA
    Abhishek Jha & Gregory Stephanopoulos
  7. Agios Pharmaceuticals, Cambridge, Massachusetts, USA
    Hua Yang, Charles Kung, Katharine E Yen, Kaiko Kunii, Francesco G Salituro, Shengfang Jin & Lenny Dang
  8. Department of Bioengineering, University of California, San Diego, California, USA
    Christian M Metallo
  9. Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, USA
    Kevin D Courtney & Matthew G Vander Heiden
  10. Department of Pathology, Children's Hospital, Boston, Massachusetts, USA
    Marian H Harris
  11. Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA
    John M Asara
  12. Department of Pharmacology, University of Toronto, Toronto, Ontario, Canada
    Hee-Won Park

Authors

  1. Dimitrios Anastasiou
  2. Yimin Yu
  3. William J Israelsen
  4. Jian-Kang Jiang
  5. Matthew B Boxer
  6. Bum Soo Hong
  7. Wolfram Tempel
  8. Svetoslav Dimov
  9. Min Shen
  10. Abhishek Jha
  11. Hua Yang
  12. Katherine R Mattaini
  13. Christian M Metallo
  14. Brian P Fiske
  15. Kevin D Courtney
  16. Scott Malstrom
  17. Tahsin M Khan
  18. Charles Kung
  19. Amanda P Skoumbourdis
  20. Henrike Veith
  21. Noel Southall
  22. Martin J Walsh
  23. Kyle R Brimacombe
  24. William Leister
  25. Sophia Y Lunt
  26. Zachary R Johnson
  27. Katharine E Yen
  28. Kaiko Kunii
  29. Shawn M Davidson
  30. Heather R Christofk
  31. Christopher P Austin
  32. James Inglese
  33. Marian H Harris
  34. John M Asara
  35. Gregory Stephanopoulos
  36. Francesco G Salituro
  37. Shengfang Jin
  38. Lenny Dang
  39. Douglas S Auld
  40. Hee-Won Park
  41. Lewis C Cantley
  42. Craig J Thomas
  43. Matthew G Vander Heiden

Contributions

D.A., Y.Y., W.J.I. and M.G.V.H. designed and coordinated the study. M.B.B., C.J.T., L.C.C., H.-W.P. and L.D. advised on various aspects of the study. J.-K.J., M.B.B., M.S., A.P.S., H.V., N.S., M.J.W., K.R.B., W.L., C.P.A., J.I., D.S.A. and C.J.T. designed and provided compounds. B.S.H., W.T., S.D. and H.-W.P. performed all structural studies. A.J. did additional structural analysis. H.Y., C.K., K.E.Y., K.K., F.G.S., S.J. and L.D. performed in vivo pharmacology and ADME studies. C.M.M., J.M.A. and G.S. did MS. M.H.H. reviewed pathology. D.A., Y.Y., W.J.I., K.R.M., B.P.F., K.D.C., S.M., T.M.K., C.K., S.Y.L., Z.R.J., S.M.D., H.R.C. and M.G.V.H. all performed experiments. D.A. and M.G.V.H. wrote the paper with substantial input from Y.Y. and W.J.I.

Corresponding author

Correspondence toMatthew G Vander Heiden.

Ethics declarations

Competing interests

L.C.C. is a founder, M.G.V.H. is a consultant and A.J., H.Y., C.K., K.E.Y., K.K., F.G.S., S.J. and L.D are employed by Agios Pharmaceuticals, a company seeking to target metabolic enzymes for cancer therapy.

Supplementary information

Rights and permissions

About this article

Cite this article

Anastasiou, D., Yu, Y., Israelsen, W. et al. Pyruvate kinase M2 activators promote tetramer formation and suppress tumorigenesis.Nat Chem Biol 8, 839–847 (2012). https://doi.org/10.1038/nchembio.1060

Download citation