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STING mediates microglial pyroptosis via interaction with NLRP3 in cerebral ischaemic stroke
Wenyu Li1, Nan Shen1, Lingqi Kong1
1Department of Neurology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, China.
Background:
Ischaemia-evoked neuroinflammation is a critical pathogenic event following ischaemic stroke. Gasdermin D (GSDMD)-associated pyroptosis represents a type of inflammation-associated programmed cell death, which can exacerbate neuroinflammatory responses and brain damage. Stimulator of interferon genes (STING) was recently described as a vital innate immune adaptor protein associated with neuroinflammation. Nevertheless, the regulatory effects of STING on microglial pyroptosis post-stroke have not been well elaborated.
Methods:
STING-knockout and wild-type (WT) mice were subjected to middle cerebral artery occlusion (MCAO). STING small interfering RNA (siRNA) was transfected into BV2 cells before oxygen-glucose deprivation/reoxygenation (OGD/R). STING-overexpressing adeno-associated virus (AAV) and NOD-like receptor family pyrin domain containing 3 (NLRP3) siRNA were administered by stereotaxic injection. 2,3,5-Triphenyl tetrazolium chloride (TTC) staining, TdT-mediated dUTP nick end labeling (TUNEL) staining, Fluoro-Jade C (FJC) staining, neurobehavioural tests, immunohistochemistry, cytokine antibody array assay, transmission electron microscopy, immunoblot, Enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (qRT-PCR) were carried out. Co-immunoprecipitation assays were used to investigate the interplay between STING and NLRP3.
Results:
STING expression was increased after MCAO and mainly detected on microglia. STING deletion alleviated brain infarction, neuronal damage and neurobehavioural impairment in mice subjected to MCAO. STING knockout suppressed microglial activation and the secretion of inflammatory chemokines, accompanied by mitigation of microglial pyroptosis. Specific upregulation of microglial STING by AAV-F4/80-STING aggravated brain injury and microglial pyroptosis. Mechanistically, co-immunoprecipitation showed that STING bound to NLRP3 in microglia. Supplementation of NLRP3 siRNA reversed AAV-F4/80-STING-induced deterioration of microglial pyroptosis.
Conclusions:
The current findings indicate that STING modulates NLRP3-mediated microglial pyroptosis following MCAO. STING may serve as a therapeutic target in neuroinflammation induced by cerebral ischaemic/reperfusion (I/R) injury.
Insights
Stimulator of interferon genes (STING) promotes neuroinflammation and brain damage after stroke by activating microglial pyroptosis. Targeting STING may offer a new therapeutic strategy for ischemic stroke treatment.
Area of Science:
- Neuroscience
- Immunology
- Pathology
Background:
- Ischaemia-evoked neuroinflammation is a key factor in stroke pathology.
- Gasdermin D (GSDMD)-associated pyroptosis exacerbates brain damage and inflammation.
- Stimulator of interferon genes (STING) is implicated in neuroinflammation, but its role in microglial pyroptosis post-stroke is unclear.
Purpose of the Study:
- To investigate the role of STING in microglial pyroptosis and its impact on brain injury following ischaemic stroke.
- To elucidate the underlying mechanism of STING-mediated microglial pyroptosis.
Main Methods:
- Middle cerebral artery occlusion (MCAO) model in STING-knockout and wild-type mice.
- In vitro studies using BV2 cells with STING siRNA and oxygen-glucose deprivation/reoxygenation (OGD/R).
- Stereotaxic delivery of STING-overexpressing AAV and NLRP3 siRNA; various assays including histology, behavior tests, and molecular analyses.
Main Results:
- STING expression increased in microglia post-MCAO, correlating with brain injury.
- STING deletion reduced infarction, neuronal damage, and neuroinflammation, mitigating microglial pyroptosis.
- STING interacted with NLRP3, and STING upregulation exacerbated pyroptosis, which was reversed by NLRP3 inhibition.
Conclusions:
- STING plays a critical role in modulating NLRP3-mediated microglial pyroptosis after ischaemic stroke.
- STING represents a potential therapeutic target for neuroinflammation in cerebral ischaemic/reperfusion injury.
