MTH1 Inhibition Eradicates Cancer by Preventing Sanitation of the DNTP Pool

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.

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References

Rai P, Young J, Burton D, Giribaldi M, Onder T, Weinberg R . Enhanced elimination of oxidized guanine nucleotides inhibits oncogenic RAS-induced DNA damage and premature senescence. Oncogene. 2010; 30(12):1489-96. DOI: 10.1038/onc.2010.520. View

. Measurement of DNA oxidation in human cells by chromatographic and enzymic methods. Free Radic Biol Med. 2003; 34(8):1089-99. DOI: 10.1016/s0891-5849(03)00041-8. View

Murshudov G, Vagin A, Dodson E . Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr D Biol Crystallogr. 1997; 53(Pt 3):240-55. DOI: 10.1107/S0907444996012255. View

Farmer H, McCabe N, Lord C, Tutt A, Johnson D, Richardson T . Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature. 2005; 434(7035):917-21. DOI: 10.1038/nature03445. View

Hahn W, Counter C, Lundberg A, Beijersbergen R, Brooks M, Weinberg R . Creation of human tumour cells with defined genetic elements. Nature. 1999; 400(6743):464-8. DOI: 10.1038/22780. View

Topal M, Baker M . DNA precursor pool: a significant target for N-methyl-N-nitrosourea in C3H/10T1/2 clone 8 cells. Proc Natl Acad Sci U S A. 1982; 79(7):2211-5. PMC: 346161. DOI: 10.1073/pnas.79.7.2211. View

German P, Szaniszlo P, Hajas G, Radak Z, Bacsi A, Hazra T . Activation of cellular signaling by 8-oxoguanine DNA glycosylase-1-initiated DNA base excision repair. DNA Repair (Amst). 2013; 12(10):856-63. PMC: 3797267. DOI: 10.1016/j.dnarep.2013.06.006. View

Molina D, Jafari R, Ignatushchenko M, Seki T, Larsson E, Dan C . Monitoring drug target engagement in cells and tissues using the cellular thermal shift assay. Science. 2013; 341(6141):84-7. DOI: 10.1126/science.1233606. View

Zhang Y, Du Y, Le W, Wang K, Kieffer N, Zhang J . Redox control of the survival of healthy and diseased cells. Antioxid Redox Signal. 2011; 15(11):2867-908. DOI: 10.1089/ars.2010.3685. View

10.

Waters N, Jones R, Williams G, Sohal B . Validation of a rapid equilibrium dialysis approach for the measurement of plasma protein binding. J Pharm Sci. 2008; 97(10):4586-95. DOI: 10.1002/jps.21317. View

11.

OBrien J, Wilson I, Orton T, Pognan F . Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem. 2000; 267(17):5421-6. DOI: 10.1046/j.1432-1327.2000.01606.x. View

12.

Druker B, Tamura S, Buchdunger E, Ohno S, Segal G, Fanning S . Effects of a selective inhibitor of the Abl tyrosine kinase on the growth of Bcr-Abl positive cells. Nat Med. 1996; 2(5):561-6. DOI: 10.1038/nm0596-561. View

13.

Alexandrov L, Nik-Zainal S, Wedge D, Aparicio S, Behjati S, Biankin A . Signatures of mutational processes in human cancer. Nature. 2013; 500(7463):415-21. PMC: 3776390. DOI: 10.1038/nature12477. View

14.

Baykov A, Evtushenko O, Avaeva S . A malachite green procedure for orthophosphate determination and its use in alkaline phosphatase-based enzyme immunoassay. Anal Biochem. 1988; 171(2):266-70. DOI: 10.1016/0003-2697(88)90484-8. View

15.

Hubatsch I, Ragnarsson E, Artursson P . Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers. Nat Protoc. 2007; 2(9):2111-9. DOI: 10.1038/nprot.2007.303. View

16.

Schultz L, Chehab N, Malikzay A, Halazonetis T . p53 binding protein 1 (53BP1) is an early participant in the cellular response to DNA double-strand breaks. J Cell Biol. 2001; 151(7):1381-90. PMC: 2150674. DOI: 10.1083/jcb.151.7.1381. View

17.

De Luca G, Ventura I, Sanghez V, Russo M, Ajmone-Cat M, Cacci E . Prolonged lifespan with enhanced exploratory behavior in mice overexpressing the oxidized nucleoside triphosphatase hMTH1. Aging Cell. 2013; 12(4):695-705. DOI: 10.1111/acel.12094. View

18.

Rai P, Onder T, Young J, McFaline J, Pang B, Dedon P . Continuous elimination of oxidized nucleotides is necessary to prevent rapid onset of cellular senescence. Proc Natl Acad Sci U S A. 2009; 106(1):169-74. PMC: 2629241. DOI: 10.1073/pnas.0809834106. View

19.

Luo M, He H, Kelley M, Georgiadis M . Redox regulation of DNA repair: implications for human health and cancer therapeutic development. Antioxid Redox Signal. 2009; 12(11):1247-69. PMC: 2864659. DOI: 10.1089/ars.2009.2698. View

20.

Fujikawa K, Kamiya H, Yakushiji H, Fujii Y, Nakabeppu Y, Kasai H . The oxidized forms of dATP are substrates for the human MutT homologue, the hMTH1 protein. J Biol Chem. 1999; 274(26):18201-5. DOI: 10.1074/jbc.274.26.18201. View