Disulfiram inhibits bacterial growth by inducing zinc-dependent reactive oxygen species

IntroductionThe discovery of novel antimicrobial mechanisms among existing clinical drugs is urgently needed. Disulfiram, an FDA-approved treatment for alcohol dependence, exhibits broad-spectrum antibacterial effects. However, its mechanism of action remains incompletely understood.MethodsThe antim...

Full description

Saved in:
Bibliographic Details
Main Authors: Qinyu Luo, Zehua Wu, Yihang Pan, Yan Zhang
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Microbiology
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1619416/full
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850102398674010112
author Qinyu Luo
Zehua Wu
Yihang Pan
Yan Zhang
author_facet Qinyu Luo
Zehua Wu
Yihang Pan
Yan Zhang
author_sort Qinyu Luo
collection DOAJ
description IntroductionThe discovery of novel antimicrobial mechanisms among existing clinical drugs is urgently needed. Disulfiram, an FDA-approved treatment for alcohol dependence, exhibits broad-spectrum antibacterial effects. However, its mechanism of action remains incompletely understood.MethodsThe antimicrobial activity of disulfiram was assessed using bacterial growth curves and colony-forming unit assays. Cytotoxicity was evaluated via propidium iodide staining and flow cytometry. Synergy with polymyxins or kanamycin was examined using checkerboard assays. RNA-seq was performed on disulfiram-treated E. coli, and differentially expressed genes were analyzed using the R package limma. Intracellular reactive oxygen species (ROS) levels were measured with fluorescent probes and flow cytometry.ResultsDisulfiram exhibited bacteriostatic, but not bactericidal, effects against E. coli and S. aureus. However, it significantly enhanced the bactericidal activity of colistin or kanamycin, both in vitro and in a murine E. coli infection model. Transcriptomic analysis revealed oxidative stress and zinc-related responses in disulfiram-treated E. coli. The bacteriostatic effects were reversed by the ROS scavenger N-acetyl-l-cysteine and zinc chelators, whereas zinc supplementation enhanced ROS production and growth inhibition.DiscussionThis study identifies a zinc-dependent ROS-mediated mechanism underlying the bacteriostatic activity of disulfiram. Although the in vivo concentrations of disulfiram during standard therapy are below its MIC, its synergistic effect with colistin suggests clinical relevance as an adjuvant. Disulfiram-induced redox stress and zinc modulation likely compromise bacterial antioxidant defenses and membrane integrity. These findings support further investigation of dithiocarbamate-based compounds as potential adjuvants or scaffolds for novel antimicrobial development.
format Article
id doaj-art-318763954b884459a867157f94346a28
institution DOAJ
issn 1664-302X
language English
publishDate 2025-07-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Microbiology
spelling doaj-art-318763954b884459a867157f94346a282025-08-20T02:39:45ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-07-011610.3389/fmicb.2025.16194161619416Disulfiram inhibits bacterial growth by inducing zinc-dependent reactive oxygen speciesQinyu LuoZehua WuYihang PanYan ZhangIntroductionThe discovery of novel antimicrobial mechanisms among existing clinical drugs is urgently needed. Disulfiram, an FDA-approved treatment for alcohol dependence, exhibits broad-spectrum antibacterial effects. However, its mechanism of action remains incompletely understood.MethodsThe antimicrobial activity of disulfiram was assessed using bacterial growth curves and colony-forming unit assays. Cytotoxicity was evaluated via propidium iodide staining and flow cytometry. Synergy with polymyxins or kanamycin was examined using checkerboard assays. RNA-seq was performed on disulfiram-treated E. coli, and differentially expressed genes were analyzed using the R package limma. Intracellular reactive oxygen species (ROS) levels were measured with fluorescent probes and flow cytometry.ResultsDisulfiram exhibited bacteriostatic, but not bactericidal, effects against E. coli and S. aureus. However, it significantly enhanced the bactericidal activity of colistin or kanamycin, both in vitro and in a murine E. coli infection model. Transcriptomic analysis revealed oxidative stress and zinc-related responses in disulfiram-treated E. coli. The bacteriostatic effects were reversed by the ROS scavenger N-acetyl-l-cysteine and zinc chelators, whereas zinc supplementation enhanced ROS production and growth inhibition.DiscussionThis study identifies a zinc-dependent ROS-mediated mechanism underlying the bacteriostatic activity of disulfiram. Although the in vivo concentrations of disulfiram during standard therapy are below its MIC, its synergistic effect with colistin suggests clinical relevance as an adjuvant. Disulfiram-induced redox stress and zinc modulation likely compromise bacterial antioxidant defenses and membrane integrity. These findings support further investigation of dithiocarbamate-based compounds as potential adjuvants or scaffolds for novel antimicrobial development.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1619416/fullEscherichia colicolistinzincROSdisulfiram
spellingShingle Qinyu Luo
Zehua Wu
Yihang Pan
Yan Zhang
Disulfiram inhibits bacterial growth by inducing zinc-dependent reactive oxygen species
Frontiers in Microbiology
Escherichia coli
colistin
zinc
ROS
disulfiram
title Disulfiram inhibits bacterial growth by inducing zinc-dependent reactive oxygen species
title_full Disulfiram inhibits bacterial growth by inducing zinc-dependent reactive oxygen species
title_fullStr Disulfiram inhibits bacterial growth by inducing zinc-dependent reactive oxygen species
title_full_unstemmed Disulfiram inhibits bacterial growth by inducing zinc-dependent reactive oxygen species
title_short Disulfiram inhibits bacterial growth by inducing zinc-dependent reactive oxygen species
title_sort disulfiram inhibits bacterial growth by inducing zinc dependent reactive oxygen species
topic Escherichia coli
colistin
zinc
ROS
disulfiram
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1619416/full
work_keys_str_mv AT qinyuluo disulfiraminhibitsbacterialgrowthbyinducingzincdependentreactiveoxygenspecies
AT zehuawu disulfiraminhibitsbacterialgrowthbyinducingzincdependentreactiveoxygenspecies
AT yihangpan disulfiraminhibitsbacterialgrowthbyinducingzincdependentreactiveoxygenspecies
AT yanzhang disulfiraminhibitsbacterialgrowthbyinducingzincdependentreactiveoxygenspecies