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RORα-activated mitophagy attenuating hypoxic-ischemic encephalopathy via suppression of microglial cGAS-STING axis
Lei Song1, Haiyan Shen1, Fei Hong1
1Department of Pediatrics, Nantong First People's Hospital (Second Affiliated Hospital of Nantong University), Nantong, Jiangsu, China.
Introduction:
Hypoxic-ischemic encephalopathy (HIE) involves neuroinflammation driven by microglial activation, yet regulatory mechanisms remain poorly defined. This study investigates how Retinoic Acid Receptor-Related Orphan Receptor Alpha (RORα) modulates mitophagy to suppress mtDNA-cGAS-STING-NLRP3 signaling in aging microglia, offering therapeutic potential for HIE.
Methods:
A multi-omics approach combining single-cell RNA sequencing (scRNA-seq) of an HIE rat model, Weighted Gene Co-Expression Network Analysis (WGCNA), and LASSO regression identified RORα as a pivotal regulator. In vivo and in vitro HIE models with RORα overexpression were assessed via behavioral tests (morris water maze, tail suspension), reactive oxygen species (ROS) quantification, and molecular profiling (RT-qPCR, Western Blot, ELISA). Mitophagy inhibitor 3-MA was used to validate pathway dependence.
Results:
Multi-omics integration revealed RORα as a hub gene linked to inflammatory and metabolic pathways. RORα activation enhanced mitophagy, reducing mtDNA leakage by 43% and cGAS-STING activity by 68%, which suppressed NLRP3 inflammasome activation (p < 0.01). This correlated with improved cognitive/motor function in HIE rats (p < 0.05) and attenuated ROS/IL-1β levels. Critically, 3-MA reversed RORα's anti-inflammatory effects, confirming mitophagy dependence.
Conclusion:
RORα alleviates HIE by resolving microglial neuroinflammation through mitophagic inhibition of mtDNA-cGAS-STING-NLRP3 signaling. These findings position RORα as a novel therapeutic target for HIE, bridging mitochondrial quality control and neuroimmunology.
Insights
Retinoic Acid Receptor-Related Orphan Receptor Alpha (RORα) resolves neuroinflammation in hypoxic-ischemic encephalopathy (HIE) by enhancing mitophagy. This suppresses harmful signaling pathways, offering a new therapeutic strategy for HIE.
Area of Science:
- Neuroimmunology
- Mitochondrial Biology
- Neuroinflammation
Background:
- Hypoxic-ischemic encephalopathy (HIE) involves microglial activation and neuroinflammation.
- Regulatory mechanisms of microglial activation in HIE are not fully understood.
- Aging microglia present unique challenges in HIE pathogenesis.
Purpose of the Study:
- Investigate the role of Retinoic Acid Receptor-Related Orphan Receptor Alpha (RORα) in modulating mitophagy.
- Determine if RORα can suppress mtDNA-cGAS-STING-NLRP3 signaling in aging microglia.
- Explore RORα as a potential therapeutic target for HIE.
Main Methods:
- Utilized a multi-omics approach including scRNA-seq and WGCNA on an HIE rat model.
- Employed in vivo and in vitro HIE models with RORα overexpression.
- Assessed outcomes via behavioral tests, ROS quantification, and molecular profiling (RT-qPCR, Western Blot, ELISA).
- Validated pathway dependence using the mitophagy inhibitor 3-MA.
Main Results:
- RORα was identified as a hub gene regulating inflammatory and metabolic pathways.
- RORα activation enhanced mitophagy, reducing mtDNA leakage by 43% and cGAS-STING activity by 68%.
- RORα suppressed NLRP3 inflammasome activation, improved cognitive/motor function in HIE rats, and reduced ROS/IL-1β levels.
- Inhibition of mitophagy reversed RORα's anti-inflammatory effects, confirming pathway dependence.
Conclusions:
- RORα alleviates HIE by resolving microglial neuroinflammation via mitophagy-mediated inhibition of mtDNA-cGAS-STING-NLRP3 signaling.
- RORα represents a novel therapeutic target for HIE.
- This study bridges mitochondrial quality control and neuroimmunology in the context of HIE.
