• A Corrigendum to this article was published on 26 April 2017

This article has been updated


Cancers have dysfunctional redox regulation resulting in reactive oxygen species production, damaging both DNA and free dNTPs. The MTH1 protein sanitizes oxidized dNTP pools to prevent incorporation of damaged bases during DNA replication. Although MTH1 is non-essential in normal cells, we show that cancer cells require MTH1 activity to avoid incorporation of oxidized dNTPs, resulting in DNA damage and cell death. We validate MTH1 as an anticancer target in vivo and describe small molecules TH287 and TH588 as first-in-class nudix hydrolase family inhibitors that potently and selectively engage and inhibit the MTH1 protein in cells. Protein co-crystal structures demonstrate that the inhibitors bind in the active site of MTH1. The inhibitors cause incorporation of oxidized dNTPs in cancer cells, leading to DNA damage, cytotoxicity and therapeutic responses in patient-derived mouse xenografts. This study exemplifies the non-oncogene addiction concept for anticancer treatment and validates MTH1 as being cancer phenotypic lethal.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Change history

  • 10 April 2014

    Asterisks were missing in Fig. 5c and have now been added.


Primary accessions


  1. 1.

    et al. Effects of a selective inhibitor of the Abl tyrosine kinase on the growth of Bcr-Abl positive cells. Nature Med. 2, 561–566 (1996)

  2. 2.

    et al. Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose)polymerase. Nature 434, 913–917 (2005)

  3. 3.

    et al. Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature 434, 917–921 (2005)

  4. 4.

    et al. Signatures of mutational processes in human cancer. Nature 500, 415–421 (2013)

  5. 5.

    et al. Redox control of the survival of healthy and diseased cells. Antioxid. Redox Signal. 15, 2867–2908 (2011)

  6. 6.

    , , & Redox regulation of DNA repair: implications for human health and cancer therapeutic development. Antioxid. Redox Signal. 12, 1247–1269 (2010)

  7. 7.

    & DNA precursor pool: a significant target for N-methyl-N-nitrosourea in C3H/10T1/2 clone 8 cells. Proc. Natl Acad. Sci. USA 79, 2211–2215 (1982)

  8. 8.

    et al. Cloning and expression of cDNA for a human enzyme that hydrolyzes 8-oxo-dGTP, a mutagenic substrate for DNA synthesis. J. Biol. Chem. 268, 23524–23530 (1993)

  9. 9.

    et al. Two distinct pathways of cell death triggered by oxidative damage to nuclear and mitochondrial DNAs. EMBO J. 27, 421–432 (2008)

  10. 10.

    et al. Oxidation of mitochondrial deoxynucleotide pools by exposure to sodium nitroprusside induces cell death. DNA Repair (Amst.) 7, 418–430 (2008)

  11. 11.

    et al. The oxidized deoxynucleoside triphosphate pool is a significant contributor to genetic instability in mismatch repair-deficient cells. Mol. Cell. Biol. 24, 465–474 (2004)

  12. 12.

    et al. Enhanced elimination of oxidized guanine nucleotides inhibits oncogenic RAS-induced DNA damage and premature senescence. Oncogene 30, 1489–1496 (2011)

  13. 13.

    , & Analysis of MTH1 gene function in mice with targeted mutagenesis. Mutat. Res. 477, 71–78 (2001)

  14. 14.

    , , & p53 binding protein 1 (53BP1) is an early participant in the cellular response to DNA double-strand breaks. J. Cell Biol. 151, 1381–1390 (2000)

  15. 15.

    Measurement of DNA oxidation in human cells by chromatographic and enzymic methods. Free Radic. Biol. Med. 34, 1089–1099 (2003)

  16. 16.

    et al. An oxidized purine nucleoside triphosphatase, MTH1, suppresses cell death caused by oxidative stress. J. Biol. Chem. 278, 37965–37973 (2003)

  17. 17.

    et al. Rb-mediated heterochromatin formation and silencing of E2F target genes during cellular senescence. Cell 113, 703–716 (2003)

  18. 18.

    et al. Monitoring drug target engagement in cells and tissues using the cellular thermal shift assay. Science 341, 84–87 (2013)

  19. 19.

    et al. Crystal structure of human MTH1 and the 8-oxo-dGMP product complex. FEBS Lett. 585, 2617–2621 (2011)

  20. 20.

    et al. A functional analysis of the DNA glycosylase activity of mouse MUTYH protein excising 2-hydroxyadenine opposite guanine in DNA. Nucleic Acids Res. 33, 672–682 (2005)

  21. 21.

    et al. Activation of cellular signaling by 8-oxoguanine DNA glycosylase-1-initiated DNA base excision repair. DNA Repair (Amst.) 12, 856–863 (2013)

  22. 22.

    et al. Creation of human tumour cells with defined genetic elements. Nature 400, 464–468 (1999)

  23. 23.

    Substrate ambiguity among the nudix hydrolases: biologically significant, evolutionary remnant, or both? Cell. Mol. Life Sci. 70, 373–385 (2013)

  24. 24.

    & MutT protein specifically hydrolyses a potent mutagenic substrate for DNA synthesis. Nature 355, 273–275 (1992)

  25. 25.

    et al. The oxidized forms of dATP are substrates for the human MutT homologue, the hMTH1 protein. J. Biol. Chem. 274, 18201–18205 (1999)

  26. 26.

    et al. Contribution of hMTH1 to the maintenance of 8-oxoguanine levels in lung DNA of non-small-cell lung cancer patients. J. Natl Cancer Inst. 97, 384–395 (2005)

  27. 27.

    , , , & ATM activation by oxidative stress. Science 330, 517–521 (2010)

  28. 28.

    et al. Stereospecific targeting of MTH1 by (S)-crizotinib as an anticancer strategy. Nature (this issue)

  29. 29.

    et al. Prolonged lifespan with enhanced exploratory behavior in mice overexpressing the oxidized nucleoside triphosphatase hMTH1. Aging Cell 12, 695–705 (2013)

  30. 30.

    et al. A role for oxidized DNA precursors in Huntington's disease-like striatal neurodegeneration. PLoS Genet. 4, e1000266 (2008)

  31. 31.

    et al. Ogg1 knockout-associated lung tumorigenesis and its suppression by Mth1 gene disruption. Cancer Res. 63, 902–905 (2003)

  32. 32.

    et al. Defects in homologous recombination repair in mismatch-repair-deficient tumour cell lines. Hum. Mol. Genet. 11, 2189–2200 (2002)

  33. 33.

    et al. Continuous elimination of oxidized nucleotides is necessary to prevent rapid onset of cellular senescence. Proc. Natl Acad. Sci. USA 106, 169–174 (2009)

  34. 34.

    , , & Inducible and reversible gene silencing by stable integration of an shRNA-encoding lentivirus in transgenic rats. Proc. Natl Acad. Sci. USA 105, 18507–18512 (2008)

  35. 35.

    , , & Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur. J. Biochem. 267, 5421–5426 (2000)

  36. 36.

    , , & Direct detection of 8-oxodeoxyguanosine and 8-oxoguanine by avidin and its analogues. Anal. Biochem. 255, 20–31 (1998)

  37. 37.

    , , , & Protein carbonyl groups as biomarkers of oxidative stress. Clin. Chim. Acta 329, 23–38 (2003)

  38. 38.

    , & A malachite green procedure for orthophosphate determination and its use in alkaline phosphatase-based enzyme immunoassay. Anal. Biochem. 171, 266–270 (1988)

  39. 39.

    & Cyclocondensations of 3-(R2-amino)-3-methylthio-1-R1-propenones with 2-aminoazoles. Ukr. Khim. Zh. 74, 126–128 (2008)

  40. 40.

    Xds. Acta Crystallogr. D 66, 125–132 (2010)

  41. 41.

    , & Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr. D 53, 240–255 (1997)

  42. 42.

    & Coot: model-building tools for molecular graphics. Acta Crystallogr. D 60, 2126–2132 (2004)

  43. 43.

    , , & Validation of a rapid equilibrium dialysis approach for the measurement of plasma protein binding. J. Pharm. Sci. 97, 4586–4595 (2008)

  44. 44.

    Utility of in vitro drug metabolism data in predicting in vivo metabolic clearance. Biochem. Pharmacol. 47, 1469–1479 (1994)

  45. 45.

    et al. Introduction to in vitro estimation of metabolic stability and drug interactions of new chemical entities in drug discovery and development. Pharmacol. Rep. 58, 453–472 (2006)

  46. 46.

    , & Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers. Nature Protocols 2, 2111–2119 (2007)

Download references


We thank scientists at BESSY, ESRF, Max-Lab and the Swiss Light Source for structural biology data collection support, PSF for protein purification, GE Healthcare for instrument access, HTSRC (Rockefeller University) for iTC200 calorimeter access, S. Nordstrand for animal support, and L. Ny, U. Stierner and J. Mattsson for discussions. The Helleday Laboratory is primarily funded by the Torsten and Ragnar Söderberg Foundation (T.H.). This project is primarily supported by The Knut and Alice Wallenberg Foundation. Further support was received from the Swedish Research Council (T.H., A.J.J., P.A., P.S., J.A.N.), the European Research Council (T.H.), Swedish Cancer Society (T.H., J.A.N.), the Swedish Children’s Cancer Foundation (T.H.), AFA insurance (T.H.), the Swedish Pain Relief Foundation (T.H.), The Cancer Society in Stockholm (T.H.), the Wenner-Gren Foundations (P.S.), the Swedish Foundation for Strategic Research (P.S.), the Dutch Cancer Society (B.E.), EMBO LTF (R.B.), Region Västra Götaland (J.A.N.), BioCARE (J.A.N.), the Swiss National Science Foundation (F.Z.G.) and the Nicholson Exchange Program (T.L.). Chemical Biology Consortium Sweden (CBCS) is primarily funded by the Swedish Research Council. CBCS acknowledge Swedish Orphan Biovitrum for their donation of a small-molecule infrastructure including a compound collection.

Author information

Author notes

    • Helge Gad
    • , Tobias Koolmeister
    • , Ann-Sofie Jemth
    • , Saeed Eshtad
    • , Sylvain A. Jacques
    •  & Cecilia E. Ström

    These authors contributed equally to this work.

    • Bastiaan Evers
    •  & Tatjana Djureinovic

    Present addresses: Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, 1006 Amsterdam, The Netherlands (B.E.); Department of Immunology, Genetics, and Pathology, Uppsala University, S-751 23 Uppsala, Sweden (T.D.).


  1. Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 21 Stockholm, Sweden

    • Helge Gad
    • , Tobias Koolmeister
    • , Ann-Sofie Jemth
    • , Saeed Eshtad
    • , Sylvain A. Jacques
    • , Cecilia E. Ström
    • , Niklas Schultz
    • , Thomas Lundbäck
    • , Camilla Göktürk
    • , Pawel Baranczewski
    • , Kia Strömberg
    • , Kumar Sanjiv
    • , Marie-Caroline Jacques-Cordonnier
    • , Matthieu Desroses
    • , Anna-Lena Gustavsson
    • , Evert J. Homan
    • , Olga Loseva
    • , Lars Bräutigam
    • , Lars Johansson
    • , Andreas Höglund
    • , Anna Hagenkort
    • , Therese Pham
    • , Mikael Altun
    • , Fabienne Z. Gaugaz
    • , Svante Vikingsson
    • , Bastiaan Evers
    • , Martin Henriksson
    • , Karl S. A. Vallin
    • , Olov A. Wallner
    • , Lars G. J. Hammarström
    • , Elisee Wiita
    • , Ingrid Almlöf
    • , Christina Kalderén
    • , Hanna Axelsson
    • , Jordi Carreras Puigvert
    • , Fredrik Jeppsson
    • , Cecilia Lundin
    • , Annika Jenmalm Jensen
    • , Martin Scobie
    • , Ulrika Warpman Berglund
    •  & Thomas Helleday
  2. Department of Biochemistry and Biophysics, Stockholm University, S-106 91 Stockholm, Sweden

    • Linda M. Svensson
    • , Ronnie P.-A. Berntsson
    • , Robert Gustafsson
    •  & Pål Stenmark
  3. Chemical Biology Consortium Sweden, Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 21 Stockholm, Sweden

    • Thomas Lundbäck
    • , Richard Svensson
    • , Anna-Lena Gustavsson
    • , Lars Johansson
    • , Lars G. J. Hammarström
    • , Hanna Axelsson
    • , Annika Jenmalm Jensen
    •  & Per Artursson
  4. Sahlgrenska Translational Melanoma Group, Sahlgrenska Cancer Center, Department of Surgery, University of Gothenburg and Sahlgrenska University Hospital, S-405 30 Gothenburg, Sweden

    • Berglind Osk Einarsdottir
    • , Roger Olofsson
    •  & Jonas A. Nilsson
  5. Department of Analytical Chemistry, Stockholm University, S-106 91 Stockholm, Sweden

    • Aljona Saleh
    •  & Ingrid Granelli
  6. Uppsala University Drug Optimization and Pharmaceutical Profiling Platform, Department of Pharmacy, Uppsala University, S-751 23 Uppsala, Sweden

    • Pawel Baranczewski
    • , Richard Svensson
    • , Fabienne Z. Gaugaz
    •  & Per Artursson
  7. Department of Genetics, Microbiology and Toxicology, Stockholm University, S-106 91 Stockholm, Sweden

    • Fredrik Johansson
    •  & Tatjana Djureinovic
  8. Clinical Pharmacology, Department of Medical and Health Sciences, Linköping University, S-58185 Linköping, Sweden

    • Svante Vikingsson
  9. Science for Life Laboratory, RNAi Cell Screening Facility, Department of Biochemistry and Biophysics, Stockholm University, S-106 91 Stockholm, Sweden

    • Maria Häggblad
    • , Ulf Martens
    •  & Bo Lundgren


  1. Search for Helge Gad in:

  2. Search for Tobias Koolmeister in:

  3. Search for Ann-Sofie Jemth in:

  4. Search for Saeed Eshtad in:

  5. Search for Sylvain A. Jacques in:

  6. Search for Cecilia E. Ström in:

  7. Search for Linda M. Svensson in:

  8. Search for Niklas Schultz in:

  9. Search for Thomas Lundbäck in:

  10. Search for Berglind Osk Einarsdottir in:

  11. Search for Aljona Saleh in:

  12. Search for Camilla Göktürk in:

  13. Search for Pawel Baranczewski in:

  14. Search for Richard Svensson in:

  15. Search for Ronnie P.-A. Berntsson in:

  16. Search for Robert Gustafsson in:

  17. Search for Kia Strömberg in:

  18. Search for Kumar Sanjiv in:

  19. Search for Marie-Caroline Jacques-Cordonnier in:

  20. Search for Matthieu Desroses in:

  21. Search for Anna-Lena Gustavsson in:

  22. Search for Roger Olofsson in:

  23. Search for Fredrik Johansson in:

  24. Search for Evert J. Homan in:

  25. Search for Olga Loseva in:

  26. Search for Lars Bräutigam in:

  27. Search for Lars Johansson in:

  28. Search for Andreas Höglund in:

  29. Search for Anna Hagenkort in:

  30. Search for Therese Pham in:

  31. Search for Mikael Altun in:

  32. Search for Fabienne Z. Gaugaz in:

  33. Search for Svante Vikingsson in:

  34. Search for Bastiaan Evers in:

  35. Search for Martin Henriksson in:

  36. Search for Karl S. A. Vallin in:

  37. Search for Olov A. Wallner in:

  38. Search for Lars G. J. Hammarström in:

  39. Search for Elisee Wiita in:

  40. Search for Ingrid Almlöf in:

  41. Search for Christina Kalderén in:

  42. Search for Hanna Axelsson in:

  43. Search for Tatjana Djureinovic in:

  44. Search for Jordi Carreras Puigvert in:

  45. Search for Maria Häggblad in:

  46. Search for Fredrik Jeppsson in:

  47. Search for Ulf Martens in:

  48. Search for Cecilia Lundin in:

  49. Search for Bo Lundgren in:

  50. Search for Ingrid Granelli in:

  51. Search for Annika Jenmalm Jensen in:

  52. Search for Per Artursson in:

  53. Search for Jonas A. Nilsson in:

  54. Search for Pål Stenmark in:

  55. Search for Martin Scobie in:

  56. Search for Ulrika Warpman Berglund in:

  57. Search for Thomas Helleday in:


T.H. devised concept and supervised the project. H.G., S.E., N.S., C.E.S., L.B., K.Sa., F.Jo., A.Hö., B.E., T.D., M.A., A.Ha., C.L., U.W.B. and T.H. designed, performed and analysed cell biology experiments. A-S.J., T.L., F.Je., O.L., K.St., T.K., M.Hä., U.M., B.L., L.J, A.J.J., E.W., C.K., I.A., S.A.J., U.W.B. and T.H. designed, performed and analysed biochemical and high-throughput experiments. L.M.S., R.P.-A.B., R.G. and P.S. designed, performed and analysed structural biology experiments. T.K., S.A.J., M.D., M.-C.J.-C., L.J., L.G.J.H., M.He, K.S.A.V., O.A.W., A.J.J., M.S. and T.H. designed, performed and analysed medicinal chemistry experiments. A.-L.G., J.C.P., E.J.H. and M.D. performed computational chemistry analysis and/or support. C.G., B.O.E., A.S., K.Sa., P.B., R.S., F.Z.G., I.G., P.A., T.P., S.V., J.A.N. and U.W.B. designed, performed and analysed ADME, pharmacology and in vivo experiments. R.O. performed surgery, and clinical follow-up, H.G., U.W.B. and T.H. wrote the paper. All authors discussed results and approved the manuscript.

Competing interests

A patent has been filed with data generated in this manuscript where T.H., M.S., T.K., S.A.J., M.D., M-C.JC. are listed as inventors.

Corresponding author

Correspondence to Thomas Helleday.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Methods

    This file contains details of the synthesis of specific MTH1 inhibitors.

About this article

Publication history






Further reading


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.