Dataset Information

Novel CH25H⁺ and OASL⁺ microglia subclusters play distinct roles in cerebral ischemic stroke.

ABSTRACT:

Background

Microglial polarization is one of the most promising therapeutic targets for multiple central nervous system (CNS) disorders, including ischemic stroke. However, detailed transcriptional alteration of microglia following cerebral ischemic stroke remains largely unclear.

Methods

Focal cerebral ischemia was induced by transient middle cerebral artery occlusion (tMCAO) for 60 min in mice. Single-cell RNA sequencing (scRNA-seq) was performed using ischemic brain tissues from tMCAO and sham mice 3 days after surgery. Ch25h^-/- mice were used to investigate the role of specific microglia subcluster on post-stroke infarct volume and neuroinflammation.

Results

We identified a relatively homeostatic subcluster with enhanced antigen processing and three "ischemic stroke associated microglia" (ISAM): MKI67⁺, CH25H⁺ and OASL⁺ subclusters. We found the MKI67⁺ subcluster undergo proliferation and differentiation into CH25H⁺ and OASL⁺ subclusters. CH25H⁺ microglia was a critical subcluster of ISAM that exhibited increased phagocytosis and neuroprotective property after stroke. Ch25h^-/- mice developed significantly increased infarct volume following ischemic stroke compared to Ch25h^+/-. Meanwhile, the OASL⁺ subcluster accumulated in the ischemic brain and was associated with the evolving of neuroinflammation after stroke, which was further aggravated in the aged mice brain.

Conclusions

Our data reveal previously unrecognized roles of the newly defined CH25H⁺ and OASL⁺ microglia subclusters following ischemic stroke, with novel insights for precise microglia modulation towards stroke therapy.

SUBMITTER: Zhang Y

PROVIDER: S-EPMC10184422 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature

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Publications

Novel CH25H<sup>+</sup> and OASL<sup>+</sup> microglia subclusters play distinct roles in cerebral ischemic stroke.

Zhang Yueman Y Guo Yunlu Y Li Ruqi R Huang Tingting T Li Yan Y Xie Wanqin W Chen Chen C Chen Weijie W Wan Jieqing J Yu Weifeng W Li Peiying P

Journal of neuroinflammation 20230515 1

<h4>Background</h4>Microglial polarization is one of the most promising therapeutic targets for multiple central nervous system (CNS) disorders, including ischemic stroke. However, detailed transcriptional alteration of microglia following cerebral ischemic stroke remains largely unclear.<h4>Methods</h4>Focal cerebral ischemia was induced by transient middle cerebral artery occlusion (tMCAO) for 60 min in mice. Single-cell RNA sequencing (scRNA-seq) was performed using ischemic brain tissues fro ...[more]

PMID: 37183260

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Project description:IntroductionThe underlying pathophysiological mechanisms of cerebral ischemia reperfusion injury (CIRI) is intricate, and current studies suggest that neuron, astrocyte, microglia, endothelial cell, and pericyte all have different phenotypic changes of specific cell types after ischemic stroke. And microglia account for the largest proportion after CIRI. Previous transcriptomic studies of ischemic stroke have typically focused on the 24 hours after CIRI, obscuring the dynamics of cellular subclusters throughout the disease process. Therefore, traditional methods for identifying cell types and their subclusters may not be sufficient to fully unveil the complexity of single-cell transcriptional profile dynamics caused by an ischemic stroke.MethodsIn this study, to explore the dynamic transcriptional profile of single cells after CIRI, we used single-cell State Transition Across-samples of RNA-seq data (scSTAR), a new bioinformatics method, to analyze the single-cell transcriptional profile of day 1, 3, and 7 of transient middle cerebral artery occlusion (tMCAO) mice. Combining our bulk RNA sequences and proteomics data, we found the importance of the integrin beta 2 (Itgb2) gene in post-modeling. And microglia of Itgb2+ and Itgb2- were clustered by the scSTAR method. Finally, the functions of the subpopulations were defined by Matescape, and three different time points after tMCAO were found to exhibit specific functions.ResultsOur analysis revealed a dynamic transcriptional profile of single cells in microglia after tMCAO and explored the important role of Itgb2 contributed to microglia by combined transcriptomics and proteomics analysis after modeling. Our further analysis revealed that the Itgb2+ microglia subcluster was mainly involved in energy metabolism, cell cycle, angiogenesis, neuronal myelin formation, and repair at 1, 3, and 7 days after tMCAO, respectively.DiscussionOur results suggested that Itgb2+ microglia act as a time-specific multifunctional immunomodulatory subcluster during CIRI, and the underlying mechanisms remain to be further investigated.

Project description:BackgroundThe pathogenic contribution of neuroinflammation to ictogenesis and epilepsy may provide a therapeutic target for reduction of seizure burden in patients that are currently underserved by traditional anti-seizure medications. The Theiler's murine encephalomyelitis virus (TMEV) model has provided important insights into the role of inflammation in ictogenesis, but questions remain regarding the relative contribution of microglia and inflammatory monocytes in this model.MethodsFemale C57BL/6 mice were inoculated by intracranial injection of 2 × 105, 5 × 104, 1.25 × 104, or 3.125 × 103 plaque-forming units (PFU) of the Daniel's strain of TMEV at 4-6 weeks of age. Infiltration of inflammatory monocytes, microglial activation, and cytokine production were measured at 24 h post-infection (hpi). Viral load, hippocampal injury, cognitive performance, and seizure burden were assessed at several timepoints.ResultsThe intensity of inflammatory infiltration and the extent of hippocampal injury induced during TMEV encephalitis scaled with the amount of infectious virus in the initial inoculum. Cognitive performance was preserved in mice inoculated with 1.25 × 104 PFU TMEV relative to 2 × 105 PFU TMEV, but peak viral load at 72 hpi was equivalent between the inocula. CCL2 production in the brain was attenuated by 90% and TNFα and IL6 production was absent in mice inoculated with 1.25 × 104 PFU TMEV. Acute infiltration of inflammatory monocytes was attenuated by more than 80% in mice inoculated with 1.25 × 104 PFU TMEV relative to 2 × 105 PFU TMEV but microglial activation was equivalent between groups. Seizure burden was attenuated and the threshold to kainic acid-induced seizures was higher in mice inoculated with 1.25 × 104 PFU TMEV but low-level behavioral seizures persisted and the EEG exhibited reduced but detectable abnormalities.ConclusionsThe size of the inflammatory monocyte response induced by TMEV scales with the amount of infectious virus in the initial inoculum, despite the development of equivalent peak infectious viral load. In contrast, the microglial response does not scale with the inoculum, as microglial hyper-ramification and increased Iba-1 expression were evident in mice inoculated with either 1.25 × 104 or 2 × 105 PFU TMEV. Inoculation conditions that drive inflammatory monocyte infiltration resulted in robust behavioral seizures and EEG abnormalities, but the low inoculum condition, associated with only microglial activation, drove a more subtle seizure and EEG phenotype.

Dataset Information

Novel CH25H⁺ and OASL⁺ microglia subclusters play distinct roles in cerebral ischemic stroke.

Background

Methods

Results

Conclusions

Publications

Novel CH25H<sup>+</sup> and OASL<sup>+</sup> microglia subclusters play distinct roles in cerebral ischemic stroke.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

Novel CH25H+ and OASL+ microglia subclusters play distinct roles in cerebral ischemic stroke.

Background

Methods

Results

Conclusions

Publications

Novel CH25H<sup>+</sup> and OASL<sup>+</sup> microglia subclusters play distinct roles in cerebral ischemic stroke.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Novel CH25H⁺ and OASL⁺ microglia subclusters play distinct roles in cerebral ischemic stroke.