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Cited by 1,145Inflammasome and immune disordersImmune Response and InflammationCell death mechanisms and regulation

FDA-approved disulfiram inhibits pyroptosis by blocking gasdermin D pore formation

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Hu, Jun Jacob, Liu, Xing, Xia, Shiyu, Zhang, Zhibin, Zhang, Ying, Zhao, Jingxia, Ruan, Jianbin, Luo, Xuemei +10 more·2020·Nature Immunology
DOI 10.1038/s41590-020-0669-6PMC PMC7316630

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Abstract

Cytosolic sensing of pathogens and damage by myeloid and barrier epithelial cells assembles large complexes called inflammasomes, which activate inflammatory caspases to process cytokines (IL-1β) and gasdermin D (GSDMD). Cleaved GSDMD forms membrane pores, leading to cytokine release and inflammatory cell death (pyroptosis). Inhibiting GSDMD is an attractive strategy to curb inflammation. Here we identify disulfiram, a drug for treating alcohol addiction, as an inhibitor of pore formation by GSDMD but not other members of the GSDM family. Disulfiram blocks pyroptosis and cytokine release in cells and lipopolysaccharide-induced septic death in mice. At nanomolar concentration, disulfiram covalently modifies human/mouse Cys191/Cys192 in GSDMD to block pore formation. Disulfiram still allows IL-1β and GSDMD processing, but abrogates pore formation, thereby preventing IL-1β release and pyroptosis. The role of disulfiram in inhibiting GSDMD provides new therapeutic indications for repurposing this safe drug to counteract inflammation, which contributes to many human diseases.

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  • At nanomolar concentrations, DIS covalently modifies cysteine 191 (human GSDMD) or cysteine 192 (mouse GSDMD), permitting upstream processing but blocking pore formation, thereby preventing IL-1β release and pyroptosis.

    “At nanomolar concentrations, DIS covalently modifies cysteine 191 (human GSDMD) or cysteine 192 (mouse GSDMD). This permits upstream processing of c) and GSDMD but blocks pore formation, thereby preventing IL-1β release and pyroptosis.117”

    From Off-label and repurposed use of disulfiram beyond alcohol dependence: A systematic review of clinical and preclinical evidence (2026) · cites this paper as [117]

    The Results sections on Cys191 and inflammasome processing report covalent modification of human GSDMD Cys191, identify mouse Cys192 as the corresponding residue, and show that disulfiram blocks pore formation and IL-1β release without apparently blocking upstream proteolytic processing. The nanomolar qualifier needs qualification: the Results report an IC50 of 0.30 μM for liposome leakage, but cellular pyroptosis IC50 values of 7.7–10.3 μM without added Cu(II), falling to 0.41 μM with Cu(II). Our copy directly demonstrates covalent modification of human Cys191, not mouse Cys192. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • By targeting gasdermin D-driven inflammation, DIS selectively inhibits pore formation by GSDMD, sparing other gasdermin family members.

    “By targeting gasdermin D (GSDMD)-driven inflammation, DIS offers promise for repurposing against diverse inflammatory diseases, as it selectively inhibits pore formation by GSDMD, sparing other gasdermin family members.”

    From Off-label and repurposed use of disulfiram beyond alcohol dependence: A systematic review of clinical and preclinical evidence (2026) · cites this paper as [117]

    The Results section, “Liposome leakage inhibition is mediated primarily by direct GSDMD pore formation” (page unavailable), reports that disulfiram inhibits GSDMD pore formation but not pore formation by pre-cleaved GSDMA3. That supports selectivity relative to GSDMA3, not the broader claim that it spares other gasdermin family members generally; the paper itself says the result *suggests* it does not inhibit the entire family. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • DIS blocks pyroptosis and cytokine release in cells and prevents lipopolysaccharide-induced septic death in mice.

    “Blocks pyroptosis and cytokine release in cells and prevents lipopolysaccharide (LPS)-induced septic death in mice.”

    From Off-label and repurposed use of disulfiram beyond alcohol dependence: A systematic review of clinical and preclinical evidence (2026) · cites this paper as [117]

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Research works printed but not linked: 37

  1. [1]Rathinam VA, Vanaja SK & Fitzgerald KA Regulation of inflammasome signaling. Nat Immunol 13, 333– 332 (2012). 22430786 PMC3523703
  2. [3]Broz P & Dixit VM Inflammasomes: mechanism of assembly, regulation and signalling. Nat Rev Immunol 16, 407– 420 (2016). 27291964
  3. [4]Kayagaki N Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. Nature 526, 666– 671 (2015). 26375259
  4. [5]Shi J Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. Nature 526, 660– 665 (2015). 26375003
  5. [6]He WT Gasdermin D is an executor of pyroptosis and required for interleukin-1beta secretion. Cell Res 25, 1285– 1298 (2015). 26611636 PMC4670995
  6. [7]Aglietti RA GsdmD p30 elicited by caspase-11 during pyroptosis forms pores in membranes. Proc Natl Acad Sci U S A 113, 7858– 7863 (2016). 27339137 PMC4948338
  7. [8]Sborgi L GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death. EMBO J 35, 1766– 1778 (2016). 27418190 PMC5010048
  8. [9]Ding J Pore-forming activity and structural autoinhibition of the gasdermin family. Nature 535, 111– 116 (2016). 27281216
  9. [11]Chen X Pyroptosis is driven by non-selective gasdermin-D pore and its morphology is different from MLKL channel-mediated necroptosis. Cell Res 26, 1007– 1020 (2016). 27573174 PMC5034106
  10. [12]Russo HM Active Caspase-1 Induces Plasma Membrane Pores That Precede Pyroptotic Lysis and Are Blocked by Lanthanides. J Immunol 197, 1353– 1367 (2016). 27385778 PMC4976007
  11. [13]Ruan J, Xia S, Liu X, Lieberman J & Wu H Cryo-EM structure of the gasdermin A3 membrane pore. Nature (2018). PMC6007975 29695864
  12. [15]Frangogiannis NG, Smith CW & Entman ML The inflammatory response in myocardial infarction. Cardiovasc Res 53, 31– 47 (2002). 11744011
  13. [16]Hoffman HM, Wanderer AA & Broide DH Familial cold autoinflammatory syndrome: phenotype and genotype of an autosomal dominant periodic fever. J Allergy Clin Immunol 108, 615– 620 (2001). 11590390 PMC4321996
  14. [17]Muckle TJ & Wellsm. Urticaria, deafness, and amyloidosis: a new heredo-familial syndrome. Q J Med 31, 235– 248 (1962). 14476827
  15. [18]Prieur AM A chronic, infantile, neurological, cutaneous and articular (CINCA) syndrome. A specific entity analysed in 30 patients. Scand J Rheumatol Suppl 66, 57– 68 (1987). 3482735
  16. [19]Hoffman HM, Mueller JL, Broide DH, Wanderer AA & Kolodner RD Mutation of a new gene encoding a putative pyrin-like protein causes familial cold autoinflammatory syndrome and Muckle-Wells syndrome. Nat Genet 29, 301– 305 (2001). 11687797 PMC4322000
  17. [20]Aksentijevich I De novo CIAS1 mutations, cytokine activation, and evidence for genetic heterogeneity in patients with neonatal-onset multisystem inflammatory disease (NOMID): a new member of the expanding family of pyrin-associated autoinflammatory diseases. Arthritis Rheum 46, 3340– 3348 (2002). 12483741 PMC4556432
  18. [21]Masters SL Familial autoinflammation with neutrophilic dermatosis reveals a regulatory mechanism of pyrin activation. Science translational medicine 8, 332ra345 (2016). 27030597
  19. [22]Rello J, Valenzuela-Sanchez F, Ruiz-Rodriguez M & Moyano S Sepsis: A Review of Advances in Management. Adv Ther 34, 2393– 2411 (2017). 29022217 PMC5702377
  20. [23]Dellinger RP Surviving Sepsis Campaign: international guidelines for management of severe sepsis and septic shock, 2012. Intensive Care Med 39, 165– 228 (2013). 23361625 PMC7095153
  21. [24]Wright C & Moore RD Disulfiram treatment of alcoholism. Am J Med 88, 647– 655 (1990). 2189310
  22. [26]Sborgi L Structure and assembly of the mouse ASC inflammasome by combined NMR spectroscopy and cryo-electron microscopy. Proc Natl Acad Sci U S A 112, 13237– 13242 (2015). 26464513 PMC4629351
  23. [27]Zhang JH, Chung TD & Oldenburg KR A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays. J Biomol Screen 4, 67– 73 (1999). 10838414
  24. [28]Sun L Mixed lineage kinase domain-like protein mediates necrosis signaling downstream of RIP3 kinase. Cell 148, 213– 227 (2012). 22265413
  25. [29]Degterev A Identification of RIP1 kinase as a specific cellular target of necrostatins. Nat Chem Biol 4, 313– 321 (2008). 18408713 PMC5434866
  26. [30]Shen ML, Johnson KL, Mays DC, Lipsky JJ & Naylor S Determination of in vivo adducts of disulfiram with mitochondrial aldehyde dehydrogenase. Biochem Pharmacol 61, 537– 545 (2001). 11239496
  27. [31]Petersen EN The pharmacology and toxicology of disulfiram and its metabolites. Acta Psychiatr Scand Suppl 369, 7– 13 (1992). 1471556
  28. [32]Nobel CS, Kimland M, Nicholson DW, Orrenius S & Slater AF Disulfiram is a potent inhibitor of proteases of the caspase family. Chem Res Toxicol 10, 1319– 1324 (1997). 9437520
  29. [33]Hagn F, Nasr ML & Wagner G Assembly of phospholipid nanodiscs of controlled size for structural studies of membrane proteins by NMR. Nat Protoc 13, 79– 98 (2018). 29215632 PMC5806515
  30. [34]Castillo-Villanueva A Disulfiram as a novel inactivator of Giardia lamblia triosephosphate isomerase with antigiardial potential. Int J Parasitol Drugs Drug Resist 7, 425– 432 (2017). 29197728 PMC5727346
  31. [35]Sanchez R, Riddle M, Woo J & Momand J Prediction of reversibly oxidized protein cysteine thiols using protein structure properties. Protein Sci 17, 473– 481 (2008). 18287280 PMC2248312
  32. [36]Rathkey JK Chemical disruption of the pyroptotic pore-forming protein gasdermin D inhibits inflammatory cell death and sepsis. Sci Immunol 3 (2018). PMC6462819 30143556
  33. [37]Juliana C Anti-inflammatory compounds parthenolide and Bay 11–7082 are direct inhibitors of the inflammasome. J Biol Chem 285, 9792– 9802 (2010). 20093358 PMC2843228
  34. [38]Nair AB & Jacob S A simple practice guide for dose conversion between animals and human. J Basic Clin Pharm 7, 27– 31 (2016). 27057123 PMC4804402
  35. [40]Sollberger G Gasdermin D plays a vital role in the generation of neutrophil extracellular traps. Sci Immunol 3 (2018). 30143555
  36. [41]Cvek B Nonprofit drugs as the salvation of the world’s healthcare systems: the case of Antabuse (disulfiram). Drug Discov Today 17, 409– 412 (2012). 22192884
  37. [42]Nasr ML Covalently circularized nanodiscs for studying membrane proteins and viral entry. Nat Methods 14, 49– 52 (2017). 27869813 PMC5199620

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Authors
Hu, J. J., Liu, X., Xia, S., Zhang, Z., Zhang, Y., Zhao, J., Ruan, J., Luo, X., Lou, X., Bai, Y., Wang, J., Hollingsworth, L. R., Magupalli, V. G., Zhao, L., Luo, H. R., Kim, J., Lieberman, J., & Wu, H.
Affiliations
  • Boston Children's Hospital (US)
  • The University of Texas Medical Branch at Galveston (US)
  • Chinese Academy of Sciences (CN)
  • Harvard University (US)
Corresponding author
  • Liu, Xing
  • Lieberman, Judy
  • Wu, Hao
ORCID
Venue
Nature Immunology
Publisher
Springer Science and Business Media LLC
Published
2020-05-04
Volume
21
Issue
7
Pages
736-745
ISSN
  • 1529-2908 (ISSN-L)
  • 1529-2916
DOI
10.1038/s41590-020-0669-6
PMC
PMC7316630
OpenAlex
W3022981886
Cited by
1,145
References
7
Citations by year
2026: 186 · 2025: 212 · 2024: 196 · 2023: 192 · 2022: 205 · 2021: 110 · 2020: 34 · 2019: 8 · 2018: 1
FWCI
43.96
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Immunology (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Heart, Lung, and Blood Institute (R01HL092020, P01HL095489)
  • National Institute of Allergy and Infectious Diseases (R01AI142642, R01AI145274, R01AI123265, R01AI139914, R01AI141386)
  • Eunice Kennedy Shriver National Institute of Child Health and Human Development (DP1HD087988)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Inflammasome and immune disorders
  • Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
Keywords
disulfiram, gasdermin D, pyroptosis, pore formation, inflammasome, IL-1β release, drug repurposing, inflammation, sepsis, covalent modification, alcoholism treatment
MeSH terms
Pyroptosis, Gasdermins, Animals, Disulfiram, Drug Evaluation, Preclinical, Female, Humans, Lipopolysaccharides, Liposomes, Recombinant Proteins, Mutagenesis, Site-Directed, Spodoptera, Sepsis, Caspases, Caspase 1, Phosphate-Binding Proteins, Cell Line, Tumor, Intracellular Signaling Peptides and Proteins, Mice, Caspases, Initiator, Interleukin-1beta, High-Throughput Screening Assays, HEK293 Cells, Drug Repositioning, Caspase Inhibitors, Sf9 Cells

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