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Pyruvate kinase M2 activators promote tetramer formation and suppress tumorigenesis

An Erratum to this article was published on 26 November 2012

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Abstract

Cancer cells engage in a metabolic program to enhance biosynthesis and support cell proliferation. The regulatory properties of pyruvate kinase M2 (PKM2) influence altered glucose metabolism in cancer. The interaction of PKM2 with phosphotyrosine-containing proteins inhibits enzyme activity and increases the availability of glycolytic metabolites to support cell proliferation. This suggests that high pyruvate kinase activity may suppress tumor growth. We show that expression of PKM1, the pyruvate kinase isoform with high constitutive activity, or exposure to published small-molecule PKM2 activators inhibits the growth of xenograft tumors. Structural studies reveal that small-molecule activators bind PKM2 at the subunit interaction interface, a site that is distinct from that of the endogenous activator fructose-1,6-bisphosphate (FBP). However, unlike FBP, binding of activators to PKM2 promotes a constitutively active enzyme state that is resistant to inhibition by tyrosine-phosphorylated proteins. These data support the notion that small-molecule activation of PKM2 can interfere with anabolic metabolism.

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Figure 1: PKM1 expression in cancer cells impairs xenograft tumor growth.
Figure 2: TEPP-46 and DASA-58 isoform specificity in vitro and in cells.
Figure 3: Activators promote PKM2 tetramer formation and prevent inhibition by phosphotyrosine signaling.
Figure 4: Structural analysis of the PKM2 activator mode of action.
Figure 5: Metabolic effects of cell treatment with PKM2 activators.
Figure 6: PKM2 activators impair xenograft growth.

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  • 11 October 2012

    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.

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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.

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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.

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Correspondence to Matthew G Vander Heiden.

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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.

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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

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