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Published on: March 7, 2019
Mitochondrial ROS promote macrophage pyroptosis by inducing GSDMD oxidation
Yufang Wang1, Peiliang Shi1, Qin Chen1
1State Key Laboratory of Pharmaceutical Biotechnology, Department of Hepatopancreatobiliary Surgery, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, MOE Key Laboratory of Model Animals for Disease Study, Model Animal Research Center, Nanjing University, Nanjing 210061, China.
Abstract:
Disrupted mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) generation are often associated with macrophage pyroptosis. It remains unclear how these forms of mitochondrial dysfunction relate to inflammasome activation and gasdermin-D (Gsdmd) cleavage, two central steps of the pyroptotic process. Here, we also found MMP collapse and ROS generation induced by Nlrp3 inflammasome activation as previous studies reported. The elimination of ROS alleviated the cleavage of Gsdmd, suggesting that Gsdmd cleavage occurs downstream of ROS release. Consistent with this result, hydrogen peroxide treatment augmented the cleavage of Gsdmd by caspase-1. Indeed, four amino acid residues of Gsdmd were oxidized under oxidative stress in macrophages. The efficiency of Gsdmd cleavage by inflammatory caspase-1 was dramatically reduced when oxidative modification was blocked by mutation of these amino acid residues. These results demonstrate that Gsdmd oxidation serves as a de novo mechanism by which mitochondrial ROS promote Nlrp3 inflammasome-dependent pyroptotic cell death.
Insights
Mitochondrial reactive oxygen species (ROS) promote macrophage pyroptosis by oxidizing gasdermin-D (Gsdmd). This oxidation is essential for Gsdmd cleavage, a key step in pyroptosis, revealing a novel ROS-driven cell death pathway.
Area of Science:
- Cellular Biology
- Immunology
- Biochemistry
Background:
- Macrophage pyroptosis involves inflammasome activation and gasdermin-D (Gsdmd) cleavage.
- Mitochondrial dysfunction, including disrupted membrane potential and reactive oxygen species (ROS) generation, is linked to pyroptosis.
Purpose of the Study:
- To elucidate the relationship between mitochondrial dysfunction (MMP collapse, ROS) and inflammasome activation/Gsdmd cleavage in pyroptosis.
- To identify the role of ROS in the pyroptotic pathway.
Main Methods:
- Investigated mitochondrial membrane potential (MMP) and ROS generation in Nlrp3 inflammasome-activated macrophages.
- Assessed the effect of ROS elimination on Gsdmd cleavage.
- Utilized hydrogen peroxide treatment to study Gsdmd cleavage by caspase-1.
- Performed site-directed mutagenesis on Gsdmd to block oxidative modification of specific amino acid residues.
Main Results:
- Nlrp3 inflammasome activation induced MMP collapse and ROS generation.
- Eliminating ROS alleviated Gsdmd cleavage, indicating Gsdmd cleavage is downstream of ROS release.
- Hydrogen peroxide treatment enhanced Gsdmd cleavage by caspase-1.
- Oxidation of four specific amino acid residues in Gsdmd was observed under oxidative stress.
- Mutating these residues significantly reduced Gsdmd cleavage efficiency by caspase-1.
Conclusions:
- Mitochondrial ROS promote Nlrp3 inflammasome-dependent pyroptosis.
- Gsdmd oxidation by ROS is a novel mechanism that facilitates Gsdmd cleavage by caspase-1.
- This study reveals a direct link between mitochondrial ROS and the execution phase of pyroptosis.
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