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

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 (LPS)-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.

NETosis: Molecular Mechanisms, Role in Physiology and Pathology

“…Of great interest is the use of existing drugs for inhibition of NETosis. So, disulfiram , which is used to treat alcoholism, inhibits GSDMD activation and protects mice in the lethal LPS-induced sepsis model [ 109 ]. It should be noted that GSDMD is also a critical component of the pyroptosis program and the authors of the study attribute its effect to the prevention of macrophage pyroptosis.…”

Therapeutic modulation of inflammasome pathways

“…LDC7559 inhibited pyroptosis by the NLRP3 and AIM2 inflammasomes, 153 but other findings suggest that it may lack activity on recombinant GSDMD in the liposome leakage assay. 160 Future studies should shed light on this apparent paradox.…”

A Multi-Phenotype System to Discover Therapies for Age-Related Dysregulation of the Immune Response to Viral Infections

“…These mechanisms could be related to the recently discovered ability of disulfiram to block the final step in inflammasome-mediated pyroptosis and cytokine release. 67 Disulfiram blocks gasdermin D (GSDMD), a pore-forming protein that plays a critical role in the terminal step of inflammasome-mediated pathways by facilitating the release of proinflammatory cytokines. [68][69][70] Cleaved GSDMD similarly creates pores on the cell membrane of neutrophils to allow for the release of neutrophil extracellular traps (NETs), a process known as NETosis.…”

“…58,73,74 Disulfiram has already been shown to protect mice from lethal lipopolysaccharide-induced septic shock. 67 There is mounting evidence that targeting NETosis and inflammasome activation could be a potential treatment for severe COVID-19. Markers in the sera of patients with severe COVID-19 indicate the presence of NETs, and an autopsy of a lung specimen from a patient with COVID-19 showed extensive neutrophil infiltration.…”