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Targeted inhibition of microglial C5aR1 by PMX205 mitigates post-ischemic stroke neuroinflammation and promotes
Jie Cao1, Saisai Tian2, Zilong Deng2
1Department of Neurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China; The Third Affiliated Hospital of Soochow University, Changzhou, Jiangsu Province, China; The First People,s Hospital of Changzhou, Changzhou, Jiangsu Province, China.
Background:
Ischemic stroke constitutes the leading cause of death and disability worldwide. Post-stroke neuroinflammation, a major driver of secondary neurodegeneration, has emerged as a priority therapeutic target. Microglia serve as critical initiators of this neuroinflammatory cascade in ischemic stroke. This study employed single-cell RNA sequencing and functional experiments to identify key regulatory factors in microglia following distinct ischemic stroke subtypes, with the goal of translating these findings into therapeutic targets for clinical application.
Methods:
To identify key regulators in ischemic stroke, we mined and analyzed public single-cell RNA sequencing datasets. Two etiologically distinct stroke models were subsequently established: permanent focal ischemia via distal middle cerebral artery occlusion (dMCAO) and transient ischemia-reperfusion injury using middle cerebral artery occlusion/reperfusion (MCAO/R). C5aR1 spatiotemporal expression was quantified through immunofluorescence (cellular localization) and quantitative immunoblotting (temporal dynamics), followed by validation of PMX205-C5aR1 binding affinity via rigid-receptor molecular docking. Therapeutic assessment included acute-phase measurements (3d post-stroke): pro-inflammatory cytokines (ELISA), cerebral infarction volume (TTC staining), and co-quantification of neuronal apoptosis/viability (TUNEL/Nissl); alongside chronic functional recovery tracking (14d): motor coordination (rotarod), sensorimotor integration (adhesive removal test), anxiety-like behavior (open field exploration), and spatial working memory (Y-maze spontaneous alternation).
Results:
Bioinformatics analysis identified significant upregulation of C5aR1 in activated microglia following ischemic stroke. This finding was corroborated by spatially resolved immunofluorescence and quantitative immunoblotting; molecular docking confirmed stable PMX205-C5aR1 binding via specific hydrophobic interactions, and subsequent therapeutic intervention with PMX205 profoundly suppressed neuroinflammation (IL-1β/IL-6/TNF-α), reduced cerebral infarction, attenuated neuronal apoptosis, and reversed long-term neurological deficits.
Conclusion:
Targeted inhibition of C5aR1 by PMX205 represents a therapeutically viable strategy to attenuate neuroinflammatory cascades and improve long-term functional recovery after ischemic stroke.
Insights
Targeting C5aR1 with PMX205 reduces neuroinflammation and brain damage after ischemic stroke. This approach improves long-term functional recovery in stroke models, offering a promising therapeutic strategy.
Area of Science:
- Neuroscience
- Immunology
- Pharmacology
Background:
- Ischemic stroke is a leading global cause of death and disability.
- Post-stroke neuroinflammation drives secondary neurodegeneration, making it a key therapeutic target.
- Microglia are central to initiating the neuroinflammatory response after ischemic stroke.
Purpose of the Study:
- Identify key regulatory factors in microglia following ischemic stroke.
- Investigate the role of C5aR1 in post-stroke neuroinflammation.
- Evaluate PMX205 as a potential therapeutic agent for ischemic stroke.
Main Methods:
- Single-cell RNA sequencing and bioinformatics analysis of stroke datasets.
- Established permanent and transient focal ischemia models (dMCAO, MCAO/R).
- Quantified C5aR1 expression, validated PMX205 binding, and assessed therapeutic effects on neuroinflammation, infarct volume, apoptosis, and functional recovery.
Main Results:
- Upregulation of C5aR1 in activated microglia post-ischemic stroke.
- PMX205 binding to C5aR1 confirmed via molecular docking.
- PMX205 treatment suppressed neuroinflammation, reduced infarct size, decreased neuronal apoptosis, and improved long-term neurological deficits.
Conclusions:
- Targeted inhibition of C5aR1 by PMX205 is a viable therapeutic strategy.
- PMX205 attenuates neuroinflammatory cascades following ischemic stroke.
- This approach improves long-term functional recovery after ischemic stroke.
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