Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Disulfiram’s anti-cancer activity reflects targeting NPL4, not inhibition of aldehyde dehydrogenase

Oncogene volume 38, pages 6711–6722 (2019)Cite this article

Subjects

Abstract

Aldehyde dehydrogenase (ALDH) is a proposed biomarker and possible target to eradicate cancer stem cells. ALDH inhibition as a treatment approach is supported by anti-cancer effects of the alcohol-abuse drug disulfiram (DSF, Antabuse). Given that metabolic products of DSF, rather than DSF itself inhibit ALDH in vivo, and that DSF’s anti-cancer activity is potentiated by copper led us to investigate the relevance of ALDH as the suggested molecular cancer-relevant target of DSF. Here we show that DSF does not directly inhibit ALDH activity in diverse human cell types, while DSF’s in vivo metabolite, S-methyl-N,N-diethylthiocarbamate-sulfoxide inhibits ALDH activity yet does not impair cancer cell viability. Our data indicate that the anti-cancer activity of DSF does not involve ALDH inhibition, and rather reflects the impact of DSF’s copper-containing metabolite (CuET), that forms spontaneously in vivo and in cell culture media, and kills cells through aggregation of NPL4, a subunit of the p97/VCP segregase. We also show that the CuET-mediated, rather than any ALDH-inhibitory activity of DSF underlies the preferential cytotoxicity of DSF towards BRCA1- and BRCA2-deficient cells. These findings provide evidence clarifying the confusing literature about the anti-cancer mechanism of DSF, a drug currently tested in clinical trials for repositioning in oncology.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Marques DS, Sandrini JZ, Boyle RT, Marins LF, Trindade GS. Relationships between multidrug resistance (MDR) and stem cell markers in human chronic myeloid leukemia cell lines. Leuk Res. 2010;34:757–62.

    Article  CAS  Google Scholar 

  2. Batlle E, Clevers H. Cancer stem cells revisited. Nat Med. 2017;23:1124–34.

    Article  CAS  Google Scholar 

  3. Tirino V, Desiderio V, Paino F, De Rosa A, Papaccio F, La Noce M, et al. Cancer stem cells in solid tumors: an overview and new approaches for their isolation and characterization. FASEB J. 2013;27:13–24.

    Article  CAS  Google Scholar 

  4. Koppaka V, Thompson DC, Chen Y, Ellermann M, Nicolaou KC, Juvonen RO, et al. Aldehyde dehydrogenase inhibitors: a comprehensive review of the pharmacology, mechanism of action, substrate specificity, and clinical application. Pharm Rev. 2012;64:520–39.

    Article  CAS  Google Scholar 

  5. Pors K, Moreb JS. Aldehyde dehydrogenases in cancer: an opportunity for biomarker and drug development? Drug Disco Today. 2014;19:1953–63.

    Article  CAS  Google Scholar 

  6. Marchitti SA, Brocker C, Stagos D, Vasiliou V. Non-P450 aldehyde oxidizing enzymes: the aldehyde dehydrogenase superfamily. Expert Opin Drug Metab Toxicol. 2008;4:697–720.

    Article  CAS  Google Scholar 

  7. Brooks PJ, Theruvathu JA. DNA adducts from acetaldehyde: implications for alcohol-related carcinogenesis. Alcohol. 2005;35:187–93.

    Article  CAS  Google Scholar 

  8. van den Hoogen C, van der Horst G, Cheung H, Buijs JT, Lippitt JM, Guzmán-Ramírez N, et al. High aldehyde dehydrogenase activity identifies tumor-initiating and metastasis-initiating cells in human prostate cancer. Cancer Res. 2010;70:5163–73.

    Article  Google Scholar 

  9. Charafe-Jauffret E, Ginestier C, Iovino F, Tarpin C, Diebel M, Esterni B, et al. Aldehyde dehydrogenase 1-positive cancer stem cells mediate metastasis and poor clinical outcome in inflammatory breast cancer. Clin Cancer Res. 2010;16:45–55.

    Article  CAS  Google Scholar 

  10. Ginestier C, Hur MH, Charafe-Jauffret E, Monville F, Dutcher J, Brown M, et al. ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell. 2007;1:555–67.

    Article  CAS  Google Scholar 

  11. Yip NC, Fombon IS, Liu P, Brown S, Kannappan V, Armesilla AL, et al. Disulfiram modulated ROS–MAPK and NFκB pathways and targeted breast cancer cells with cancer stem cell-like properties. Br J Cancer. 2011;104:1564–74.

    Article  CAS  Google Scholar 

  12. Allensworth JL, Evans MK, Bertucci F, Aldrich AJ, Festa RA, Finetti P, 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;9:1155–68.

    Article  CAS  Google Scholar 

  13. Liu X, Wang L, Cui W, Yuan X, Lin L, Cao Q, et al. Targeting ALDH1A1 by disulfiram/copper complex inhibits non-small cell lung cancer recurrence driven by ALDH-positive cancer stem cells. Oncotarget. 2016;7:58516–30.

    PubMed  PubMed Central  Google Scholar 

  14. Liu P, Kumar IS, Brown S, Kannappan V, Tawari PE, Tang JZ, et al. Disulfiram targets cancer stem-like cells and reverses resistance and cross-resistance in acquired paclitaxel-resistant triple-negative breast cancer cells. Br J Cancer. 2013;109:1876–85.

    Article  CAS  Google Scholar 

  15. Liu P, Brown S, Goktug T, Channathodiyil P, Kannappan V, Hugnot J-P, et al. Cytotoxic effect of disulfiram/copper on human glioblastoma cell lines and ALDH-positive cancer-stem-like cells. Br J Cancer. 2012;107:1488–97.

    Article  CAS  Google Scholar 

  16. Liu P, Wang Z, Brown S, Kannappan V, Tawari PE, Jiang W, et al. Liposome encapsulated Disulfiram inhibits NFkappaB pathway and targets breast cancer stem cells in vitro and in vivo. Oncotarget. 2014;5:7471–85.

    PubMed  PubMed Central  Google Scholar 

  17. Jin N, Zhu X, Cheng F, Zhang L. Disulfiram/copper targets stem cell-like ALDH(+) population of multiple myeloma by inhibition of ALDH1A1 and Hedgehog pathway. J Cell Biochem. 2018;119:6882–93.

    Article  CAS  Google Scholar 

  18. Choi SA, Choi JW, Wang K-C, Phi JH, Lee JY, Park KD, et al. Disulfiram modulates stemness and metabolism of brain tumor initiating cells in atypical teratoid/rhabdoid tumors. Neuro Oncol. 2015;17:810–21.

    Article  CAS  Google Scholar 

  19. MacDonagh L, Gallagher MF, Ffrench B, Gasch C, Breen E, Gray SG, et al. Targeting the cancer stem cell marker, aldehyde dehydrogenase 1, to circumvent cisplatin resistance in NSCLC. Oncotarget. 2017;8:72544–63.

    Article  Google Scholar 

  20. Tacconi EM, Lai X, Folio C, Porru M, Zonderland G, Badie S, et al. BRCA1 and BRCA2 tumor suppressors protect against endogenous acetaldehyde toxicity. EMBO Mol Med. 2017;9:1398–414.

    Article  CAS  Google Scholar 

  21. Raha D, Wilson TR, Peng J, Peterson D, Yue P, Evangelista M, et al. The cancer stem cell marker aldehyde dehydrogenase is required to maintain a drug-tolerant tumor cell subpopulation. Cancer Res. 2014;74:3579–90.

    Article  CAS  Google Scholar 

  22. Wu L, Meng F, Dong L, Block CJ, Mitchell AV, Wu J, et al. Disulfiram and BKM120 in combination with chemotherapy impede tumor progression and delay tumor recurrence in tumor initiating cell-rich TNBC. Sci Rep. 2019;9:236.

    Article  Google Scholar 

  23. Wang N-N, Wang L-H, Li Y, Fu S-Y, Xue X, Jia L-N, et al. Targeting ALDH2 with disulfiram/copper reverses the resistance of cancer cells to microtubule inhibitors. Exp Cell Res. 2018;362:72–82.

    Article  Google Scholar 

  24. Bista R, Lee DW, Pepper OB, Azorsa DO, Arceci RJ, Aleem E. Disulfiram overcomes bortezomib and cytarabine resistance in Down-syndrome-associated acute myeloid leukemia cells. J Exp Clin Cancer Res. 2017;36:22.

    Article  Google Scholar 

  25. Lipsky JJ, Shen ML, Naylor S. In vivo inhibition of aldehyde dehydrogenase by disulfiram. Chem Biol Inter. 2001;130–132:93–102.

    Article  Google Scholar 

  26. Shen ML, Johnson KL, Mays DC, Lipsky JJ, Naylor S. Determination of in vivo adducts of disulfiram with mitochondrial aldehyde dehydrogenase. Biochem Pharm. 2001;61:537–45.

    Article  CAS  Google Scholar 

  27. Mays DC, Nelson AN, Fauq AH, Shriver ZH, Veverka KA, Naylor S, et al. S-Methyl N,N-diethylthiocarbamate sulfone, a potential metabolite of disulfiram and potent inhibitor of low Km mitochondrial aldehyde dehydrogenase. Biochem Pharm. 1995;49:693–700.

    Article  CAS  Google Scholar 

  28. Lam JP, Mays DC, Lipsky JJ. Inhibition of recombinant human mitochondrial and cytosolic aldehyde dehydrogenases by two candidates for the active metabolites of disulfiram †. Biochemistry. 1997;36:13748–54.

    Article  CAS  Google Scholar 

  29. Hart BW, Faiman MD. Bioactivation of S-methyl N,N-Diethylthiolcarbamate to S-methyl N,N-diethylthiolcarbamate sulfoxide. Biochem Pharm. 1993;46:2285–90.

    Article  CAS  Google Scholar 

  30. Yourick JJ, Faiman MD. Disulfiram metabolism as a requirement for the inhibition of rat liver mitochondrial low Km aldehyde dehydrogenase. Biochem Pharm. 1991;42:1361–6.

    Article  CAS  Google Scholar 

  31. Chen D, Cui QC, Yang H, Dou QP. 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;66:10425–33.

    Article  CAS  Google Scholar 

  32. Majera D, Skrott Z, Bouchal J, Bartkova J, Simkova D, Gachechiladze M, et al. Targeting genotoxic and proteotoxic stress-response pathways in human prostate cancer by clinically available PARP inhibitors, vorinostat and disulfiram. Prostate. 2019;79:352–62.

    Article  CAS  Google Scholar 

  33. Skrott Z, Mistrik M, Andersen KK, Friis S, Majera D, Gursky J, et al. Alcohol-abuse drug disulfiram targets cancer via p97 segregase adaptor NPL4. Nature. 2017;552:194–9.

    Article  CAS  Google Scholar 

  34. Farmer H, McCabe N, Lord CJ, Tutt ANJ, Johnson DA, Richardson TB, et al. Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature. 2005;434:917–21.

    Article  CAS  Google Scholar 

  35. Bryan N, Andrews KD, Loughran MJ, Rhodes NP, Hunt JA. Elucidating the contribution of the elemental composition of fetal calf serum to antigenic expression of primary human umbilical-vein endothelial cells in vitro. Biosci Rep. 2011;31:199–210.

    Article  CAS  Google Scholar 

  36. Zhou L, Sheng D, Wang D, Ma W, Deng Q, Deng L, et al. Identification of cancer-type specific expression patterns for active aldehyde dehydrogenase (ALDH) isoforms in ALDEFLUOR assay. Cell Biol Toxicol. 2019;35:161–77.

    Article  Google Scholar 

  37. Huddle BC, Grimley E, Buchman CD, Chtcherbinine M, Debnath B, Mehta P, et al. Structure-based optimization of a novel class of aldehyde dehydrogenase 1A (ALDH1A) subfamily-selective inhibitors as potential adjuncts to ovarian cancer chemotherapy. J Med Chem. 2018;61:8754–73.

    Article  CAS  Google Scholar 

  38. Duan J-J, Cai J, Guo Y-F, Bian X-W, Yu S-C. ALDH1A3, a metabolic target for cancer diagnosis and therapy. Int J Cancer. 2016;139:965–75.

    Article  CAS  Google Scholar 

  39. Lai C-L, Yao C-T, Chau G-Y, Yang L-F, Kuo T-Y, Chiang C-P, et al. Dominance of the inactive Asian variant over activity and protein contents of mitochondrial aldehyde dehydrogenase 2 in human liver. Alcohol Clin Exp Res. 2014;38:44–50.

    Article  CAS  Google Scholar 

  40. Chang JS, Hsiao J-R, Chen C-H. ALDH2 polymorphism and alcohol-related cancers in Asians: a public health perspective. J Biomed Sci. 2017;24:19.

    Article  Google Scholar 

  41. Suh JJ, Pettinati HM, Kampman KM, O’Brien CP. The status of disulfiram: a half of a century later. J Clin Psychopharmacol. 2006;26:290–302.

    Article  CAS  Google Scholar 

  42. Chen W, Yang W, Chen P, Huang Y, Li F. Disulfiram copper nanoparticles prepared with a stabilized metal ion ligand complex method for treating drug-resistant prostate cancers. ACS Appl Mater Interfaces. 2018;10:41118–28.

    Article  CAS  Google Scholar 

  43. Peng X, Pan Q, Zhang B, Wan S, Li S, Luo K, et al. Highly stable, coordinated polymeric nanoparticles loading copper(II) diethyldithiocarbamate for combinational chemo/chemodynamic therapy of cancer. Biomacromolecules. 2019;20:2372–83.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr. M. Tarsounas (Oxford, UK) for the human H1299 cell lines with regulatable expression of shBRCA1 and shBRCA2. The study was supported by grants from: Grant agency of Czech Rep. GACR 17–25976 S, MEYS CR (LM2015062 Czech‐BioImaging and DRO‐61989592), Internal grant of University of Palacky IGA_LF_2019_026, Cancer Research Czech Republic, Ministry of School, Education, Youth and Sports of the Czech Republic (EATRIS-CZ No. LM2015064 and ENOCH No. CZ.02.1.01/0.0/0.0/16_019/0000868), the Novo Nordisk Foundation (no. 16854), the Danish National Research Foundation (project CARD: no. DNRF125), the Danish Cancer Society (R204-A12617) the Swedish Research Council (VR-MH 2014–46602–117891–30), and the Swedish Cancer Society (no. 170176).

Author information

Author notes

  1. These authors contributed equally: Zdenek Skrott, Dusana Majera

Authors and Affiliations

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

    Zdenek Skrott, Dusana Majera, Jan Gursky, Tereza Buchtova, Marian Hajduch, Martin Mistrik & Jiri Bartek

  2. Danish Cancer Society Research Center, Copenhagen, Denmark

    Jiri Bartek

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

    Jiri Bartek

Authors
  1. Zdenek Skrott
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dusana Majera
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jan Gursky
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tereza Buchtova
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Marian Hajduch
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Martin Mistrik
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jiri Bartek
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Martin Mistrik or Jiri Bartek.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Skrott, Z., Majera, D., Gursky, J. et al. Disulfiram’s anti-cancer activity reflects targeting NPL4, not inhibition of aldehyde dehydrogenase. Oncogene 38, 6711–6722 (2019). https://doi.org/10.1038/s41388-019-0915-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-019-0915-2

This article is cited by

Search

Advanced search

Quick links