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

Jun Jacob Hu; Xing Liu; Shiyu Xia; Zhibin Zhang; Ying Zhang; Jingxia Zhao; Jianbin Ruan; Xuemei Luo; Xiwen Lou; Yang Bai; Junhong Wang; L Robert Hollingsworth; Venkat Giri Magupalli; Li Zhao; Hongbo R Luo; Justin Kim; Judy Lieberman; Hao Wu

doi:10.1038/s41590-020-0669-6

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

Nat Immunol. 2020 Jul;21(7):736-745. doi: 10.1038/s41590-020-0669-6. Epub 2020 May 4.

Authors

Jun Jacob Hu^#^{1

2}, Xing Liu^#^{3

4

5}, Shiyu Xia^{1

2}, Zhibin Zhang^{1

6}, Ying Zhang^{1

6}, Jingxia Zhao^{1

2

7}, Jianbin Ruan^{1

2

8}, Xuemei Luo⁹, Xiwen Lou¹⁰, Yang Bai¹⁰, Junhong Wang¹⁰, L Robert Hollingsworth^{1

2}, Venkat Giri Magupalli^{1

2}, Li Zhao^{11

12

13}, Hongbo R Luo^{11

12

13}, Justin Kim^{2

14}, Judy Lieberman^{15

16}, Hao Wu^{17

18}

Affiliations

¹ Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.
² Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA.
³ Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA. xingliu@ips.ac.cn.
⁴ Department of Pediatrics, Harvard Medical School, Boston, MA, USA. xingliu@ips.ac.cn.
⁵ The Center for Microbes, Development and Health, Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China. xingliu@ips.ac.cn.
⁶ Department of Pediatrics, Harvard Medical School, Boston, MA, USA.
⁷ Beijing Institute of Traditional Chinese Medicine, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing, China.
⁸ Department of Immunology, University of Connecticut Health Center, Farmington, CT, USA.
⁹ Biomolecular Resource Facility, University of Texas Medical Branch, Galveston, TX, USA.
¹⁰ The Center for Microbes, Development and Health, Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China.
¹¹ Department of Laboratory Medicine, The Stem Cell Program, Boston Children's Hospital, Boston, MA, USA.
¹² Department of Pathology, Harvard Medical School, Boston, MA, USA.
¹³ Dana-Farber/Harvard Cancer Center, Boston, MA, USA.
¹⁴ Dana-Farber Cancer Institute, Boston, MA, USA.
¹⁵ Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA. judy.lieberman@childrens.harvard.edu.
¹⁶ Department of Pediatrics, Harvard Medical School, Boston, MA, USA. judy.lieberman@childrens.harvard.edu.
¹⁷ Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA. wu@crystal.harvard.edu.
¹⁸ Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA. wu@crystal.harvard.edu.

^# Contributed equally.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Caspase 1 / genetics
Caspase 1 / metabolism
Caspase Inhibitors / pharmacology
Caspases / metabolism
Caspases, Initiator / genetics
Caspases, Initiator / metabolism
Cell Line, Tumor
Disulfiram / pharmacology*
Disulfiram / therapeutic use
Drug Evaluation, Preclinical
Drug Repositioning
Female
HEK293 Cells
High-Throughput Screening Assays
Humans
Interleukin-1beta / immunology
Interleukin-1beta / metabolism
Intracellular Signaling Peptides and Proteins / antagonists & inhibitors*
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism
Lipopolysaccharides / administration & dosage
Lipopolysaccharides / immunology
Liposomes
Mice
Mutagenesis, Site-Directed
Phosphate-Binding Proteins / antagonists & inhibitors*
Phosphate-Binding Proteins / genetics
Phosphate-Binding Proteins / metabolism
Pyroptosis / drug effects*
Pyroptosis / immunology
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Sepsis / drug therapy*
Sepsis / immunology
Sf9 Cells
Spodoptera

Substances

Caspase Inhibitors
GSDMD protein, human
Gsdmd protein, mouse
Interleukin-1beta
Intracellular Signaling Peptides and Proteins
Lipopolysaccharides
Liposomes
Phosphate-Binding Proteins
Recombinant Proteins
Casp4 protein, mouse
Caspases
Caspases, Initiator
caspase 11, human
Caspase 1
Disulfiram

Abstract

Publication types

MeSH terms

Substances

Grants and funding