Cancer incidence is rising and this global challenge is further exacerbated by tumour resistance to available medicines. A promising approach to meet the need for improved cancer treatment is drug repurposing. Here we highlight the potential for repurposing disulfiram (also known by the trade name Antabuse), an old alcohol-aversion drug that has been shown to be effective against diverse cancer types in preclinical studies. Our nationwide epidemiological study reveals that patients who continuously used disulfiram have a lower risk of death from cancer compared to those who stopped using the drug at their diagnosis. Moreover, we identify the ditiocarb-copper complex as the metabolite of disulfiram that is responsible for its anti-cancer effects, and provide methods to detect preferential accumulation of the complex in tumours and candidate biomarkers to analyse its effect on cells and tissues. Finally, our functional and biophysical analyses reveal the molecular target of disulfiram's tumour-suppressing effects as NPL4, an adaptor of p97 (also known as VCP) segregase, which is essential for the turnover of proteins involved in multiple regulatory and stress-response pathways in cells.

Barbara Ann Karmanos Cancer Institute and Department of Oncology School of Medicine Wayne State University Detroit Michigan USA

Danish Cancer Society Research Center DK 2100 Copenhagen Denmark

Department of Cell Biology and Genetics Palacky University Olomouc Czech Republic

Division of Biology and Biological Engineering Caltech Pasadena California 91125 USA

Howard Hughes Medical Institute Caltech Pasadena California 91125 USA

Institute of Biophysics and Informatics 1st Faculty of Medicine Charles University 120 00 Prague 2 Czech Republic

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacky University Olomouc Czech Republic

Kantonsspital St Gallen Department Oncology Hematology St Gallen Switzerland

Psychiatric Hospital 785 01 Šternberk Czech Republic

School of Basic Medical Sciences Affiliated Tumor Hospital of Guangzhou Medical University Guangzhou 511436 China

Science for Life Laboratory Division of Genome Biology Department of Medical Biochemistry and Biophysics Karolinska Institute Stockholm Sweden

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Cancer Discov. 2018 Feb;8(2):OF7. doi: 10.1158/2159-8290.CD-RW2017-235. PubMed

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Nat Chem Biol. 2018 Jan 16;14(2):107. doi: 10.1038/nchembio.2562. PubMed

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Transl Cancer Res. 2018 Apr;7(Suppl 4):S495-S499. doi: 10.21037/tcr.2018.03.33. PubMed

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Collins FS. Mining for therapeutic gold. Nat Rev Drug Discov. 2011;10(6):397. PubMed PMC

Pantziarka P, et al. The Repurposing Drugs in Oncology (ReDO) Project. Ecancermedicalscience. 2014;8:442. PubMed PMC

Iljin K, et al. High-throughput cell-based screening of 4910 known drugs and drug-like small molecules identifies disulfiram as an inhibitor of prostate cancer cell growth. Clin Cancer Res. 2009;15(19):6070–6078. PubMed

Cvek B. Nonprofit drugs as the salvation of the world’s healthcare systems: the case of Antabuse (disulfiram) Drug Discov Today. 2012;17(9–10):409–412. PubMed

Shen ML, et al. Determination of in vivo adducts of disulfiram with mitochondrial aldehyde dehydrogenase. Biochem Pharmacol. 2001;61(5):537–545. PubMed

Chen D, et al. Disulfiram, a clinically used anti-alcoholism drug and copper-binding agent, induces apoptotic cell death in breast cancer cultures and xenografts via inhibition of the proteasome activity. Cancer Res. 2006;66(21):10425–10433. PubMed

Zha J, et al. Disulfiram targeting lymphoid malignant cell lines via ROS-JNK activation as well as Nrf2 and NF-kB pathway inhibition. J Transl Med. 2014;12:163. PubMed PMC

Safi R, et al. Copper signaling axis as a target for prostate cancer therapeutics. Cancer Res. 2014;74(20):5819–5831. PubMed PMC

Liu P, et al. Liposome encapsulated Disulfiram inhibits NFkappaB pathway and targets breast cancer stem cells in vitro and in vivo. Oncotarget. 2014;5(17):7471–7485. PubMed PMC

Dufour P, et al. Sodium dithiocarb as adjuvant immunotherapy for high risk breast cancer: a randomized study. Biotherapy. 1993;6(1):9–12. PubMed

Yip NC, et al. Disulfiram modulated ROS-MAPK and NFkappaB pathways and targeted breast cancer cells with cancer stem cell-like properties. Br J Cancer. 2011;104(10):1564–1574. PubMed PMC

Lovborg H, et al. Inhibition of proteasome activity, nuclear factor-KappaB translocation and cell survival by the antialcoholism drug disulfiram. Int J Cancer. 2006;118(6):1577–1780. PubMed

Allensworth JL, et al. Disulfiram (DSF) acts as a copper ionophore to induce copper-dependent oxidative stress and mediate anti-tumor efficacy in inflammatory breast cancer. Mol Oncol. 2015;9(6):1155–1168. PubMed PMC

Nechushtan H, et al. A phase IIb trial assessing the addition of disulfiram to chemotherapy for the treatment of metastatic non-small cell lung cancer. Oncologist. 2015;20(4):366–367. PubMed PMC

Jin M, et al. Alcohol drinking and all cancer mortality: a meta-analysis. Ann Oncol. 2013;24(3):807–16. PubMed

Li Y, et al. Copper improves the anti-angiogenic activity of disulfiram through the EGFR/Src/VEGF pathway in gliomas. Cancer Lett. 2015;369(1):86–96. PubMed

Suzuki Y, et al. The origin of an EPR signal observed in dithiocarbamate-loaded tissues. Copper(II)-dithiocarbamate complexes account for the narrow hyperfine lines. Biochim Biophys Acta. 1997;1335(3):242–245. PubMed

Kepp O, et al. Cell death assays for drug discovery. Nat Rev Drug Discov. 2011;10(3):221–237. PubMed

Doil C, et al. RNF168 binds and amplifies ubiquitin conjugates on damaged chromosomes to allow accumulation of repair proteins. Cell. 2009;136(3):435–446. PubMed

Li JM, et al. The p97-UFD1L-NPL4 protein complex mediates cytokine-induced IkappaBalpha proteolysis. Mol Cell Biol. 2014;34(3):335–347. PubMed PMC

Chou TF, Deshaies RJ. Quantitative cell-based protein degradation assays to identify and classify drugs that target the ubiquitin-proteasome system. J Biol Chem. 2011;286(19):16546–16554. PubMed PMC

Kisselev AF, Goldberg AL. Monitoring activity and inhibition of 26S proteasomes with fluorogenic peptide substrates. Methods Enzymol. 2005;398:364–378. PubMed

Asher G, et al. Regulation of p53 stability and p53-dependent apoptosis by NADH quinone oxidoreductase 1. Proc Natl Acad Sci U S A. 2001;98(3):1188–1193. PubMed PMC

Asher G, et al. A mechanism of ubiquitin-independent proteasomal degradation of the tumor suppressors p53 and p73. Genes Dev. 2005;19(3):316–321. PubMed PMC

Verma R, et al. Role of Rpn11 metalloprotease in deubiquitination and degradation by the 26S proteasome. Science. 2002;298(5593):611–615. PubMed

Dai RM, Li CC. Valosin-containing protein is a multi-ubiquitin chain-targeting factor required in ubiquitin-proteasome degradation. Nat Cell Biol. 2001;3(8):740–744. PubMed

Alexandru G, et al. UBXD7 binds multiple ubiquitin ligases and implicates p97 in HIF1alpha turnover. Cell. 2008;134(5):804–816. PubMed PMC

Chou TF, et al. Reversible inhibitor of p97, DBeQ, impairs both ubiquitin-dependent and autophagic protein clearance pathways. Proc Natl Acad Sci U S A. 2011;108(12):4834–4839. PubMed PMC

Riemer A, et al. The p97-Ufd1-Npl4 ATPase complex ensures robustness of the G2/M checkpoint by facilitating CDC25A degradation. Cell Cycle. 2014;13(6):919–927. PubMed PMC

Radhakrishnan SK, den Besten W, Deshaies RJ. p97-dependent retrotranslocation and proteolytic processing govern formation of active Nrf1 upon proteasome inhibition. Elife. 2014;3:e01856. PubMed PMC

Meyer H, Bug M, Bremer S. Emerging functions of the VCP/p97 AAA-ATPase in the ubiquitin system. Nat Cell Biol. 2012;14(2):117–123. PubMed

Magnaghi P, et al. Covalent and allosteric inhibitors of the ATPase VCP/p97 induce cancer cell death. Nat Chem Biol. 2013;9(9):548–556. PubMed

Samali A, et al. Methods for monitoring endoplasmic reticulum stress and the unfolded protein response. Int J Cell Biol. 2010;2010:830307. PubMed PMC

Auner HW, et al. Combined inhibition of p97 and the proteasome causes lethal disruption of the secretory apparatus in multiple myeloma cells. PLoS One. 2013;8(9):e74415. PubMed PMC

Soriano GP, et al. Proteasome inhibitor-adapted myeloma cells are largely independent from proteasome activity and show complex proteomic changes, in particular in redox and energy metabolism. Leukemia. 2016;30(11):2198–2207. PubMed PMC

Lass A, et al. Analysis of Npl4 deletion mutants in mammalian cells unravels new Ufd1-interacting motifs and suggests a regulatory role of Npl4 in ERAD. Exp Cell Res. 2008;314(14):2715–2723. PubMed

Vorackova I, et al. Purification of proteins containing zinc finger domains using immobilized metal ion affinity chromatography. Protein Expr Purif. 2011;79(1):88–95. PubMed PMC

Holdgate G, et al. Biophysical methods in drug discovery from small molecule to pharmaceutical. Methods Mol Biol. 2013;1008:327–355. PubMed

Lomenick B, et al. Target identification using drug affinity responsive target stability (DARTS) Proc Natl Acad Sci U S A. 2009;106(51):21984–21989. PubMed PMC

Becker LA, et al. Therapeutic reduction of ataxin-2 extends lifespan and reduces pathology in TDP-43 mice. Nature. 2017;544(7650):367–371. PubMed PMC

Guo L, et al. A cellular system that degrades misfolded proteins and protects against neurodegeneration. Mol Cell. 2014;55(1):15–30. PubMed PMC

Kim YE, et al. Molecular chaperone functions in protein folding and proteostasis. Annu Rev Biochem. 2013;82:323–355. PubMed

Dai C, Sampson SB. HSF1: Guardian of Proteostasis in Cancer. Trends Cell Biol. 2016;26(1):17–28. PubMed PMC

Cvek B. Targeting malignancies with disulfiram (Antabuse): multidrug resistance, angiogenesis, and proteasome. Curr Cancer Drug Targets. 2011;11(3):332–337. PubMed

Deshaies RJ. Proteotoxic crisis, the ubiquitin-proteasome system, and cancer therapy. BMC Biol. 2014;12:94. PubMed PMC

Anderson DJ, et al. Targeting the AAA ATPase p97 as an Approach to Treat Cancer through Disruption of Protein Homeostasis. Cancer Cell. 2015;28(5):653–665. PubMed PMC

Cui Y, et al. High expression of valosin-containing protein predicts poor prognosis in patients with breast carcinoma. Tumour Biol. 2015;36(12):9919–9927. PubMed

Yamamoto S, et al. Expression of valosin-containing protein in colorectal carcinomas as a predictor for disease recurrence and prognosis. Clin Cancer Res. 2004;10(2):651–657. PubMed

Tsujimoto Y, et al. Elevated expression of valosin-containing protein (p97) is associated with poor prognosis of prostate cancer. Clin Cancer Res. 2004;10(9):3007–3012. PubMed

Thygesen LC, et al. Introduction to Danish (nationwide) registers on health and social issues: structure, access, legislation, and archiving. Scand J Public Health. 2011;39(7 Suppl):12–16. PubMed

Rosenbaum PR, et al. The central role of the propensity score in observational studies for causal effects. Biometrika. 1983;70(1):41–55.

R Core Team. A language and environment for statistical computing. R Foundation for Statistical Computing; Vienna, Austria: URL https://www.R-project.org/

Cvek B, et al. Ni(II), Cu(II), and Zn(II) diethyldithiocarbamate complexes show various activities against the proteasome in breast cancer cells. J Med Chem. 2008;51(20):6256–6258. PubMed PMC

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