RORα-activated mitophagy attenuating hypoxic-ischemic encephalopathy via suppression of microglial cGAS-STING axis

IntroductionHypoxic-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-NLRP...

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Main Authors: Lei Song, Haiyan Shen, Fei Hong, Weiyan Zhang, Hongyi Lu
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1592737/full
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author Lei Song
Haiyan Shen
Fei Hong
Weiyan Zhang
Hongyi Lu
author_facet Lei Song
Haiyan Shen
Fei Hong
Weiyan Zhang
Hongyi Lu
author_sort Lei Song
collection DOAJ
description IntroductionHypoxic-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.MethodsA 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.ResultsMulti-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.ConclusionRORα 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.
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spelling doaj-art-dee5141334c546f4a3a2061ea6e867c42025-08-20T03:31:33ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-07-011610.3389/fimmu.2025.15927371592737RORα-activated mitophagy attenuating hypoxic-ischemic encephalopathy via suppression of microglial cGAS-STING axisLei SongHaiyan ShenFei HongWeiyan ZhangHongyi LuIntroductionHypoxic-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.MethodsA 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.ResultsMulti-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.ConclusionRORα 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.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1592737/fullhypoxic-ischemic encephalopathyRORαmitophagyscRNA-seqneuroinflammationcGAS-STING pathway
spellingShingle Lei Song
Haiyan Shen
Fei Hong
Weiyan Zhang
Hongyi Lu
RORα-activated mitophagy attenuating hypoxic-ischemic encephalopathy via suppression of microglial cGAS-STING axis
Frontiers in Immunology
hypoxic-ischemic encephalopathy
RORα
mitophagy
scRNA-seq
neuroinflammation
cGAS-STING pathway
title RORα-activated mitophagy attenuating hypoxic-ischemic encephalopathy via suppression of microglial cGAS-STING axis
title_full RORα-activated mitophagy attenuating hypoxic-ischemic encephalopathy via suppression of microglial cGAS-STING axis
title_fullStr RORα-activated mitophagy attenuating hypoxic-ischemic encephalopathy via suppression of microglial cGAS-STING axis
title_full_unstemmed RORα-activated mitophagy attenuating hypoxic-ischemic encephalopathy via suppression of microglial cGAS-STING axis
title_short RORα-activated mitophagy attenuating hypoxic-ischemic encephalopathy via suppression of microglial cGAS-STING axis
title_sort rorα activated mitophagy attenuating hypoxic ischemic encephalopathy via suppression of microglial cgas sting axis
topic hypoxic-ischemic encephalopathy
RORα
mitophagy
scRNA-seq
neuroinflammation
cGAS-STING pathway
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1592737/full
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AT weiyanzhang roraactivatedmitophagyattenuatinghypoxicischemicencephalopathyviasuppressionofmicroglialcgasstingaxis
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