Integrative multi-omics analysis identifies coronin 1C as a potential mediator linking circadian rhythm disruption to neuroimmune dysregulation in ischemic stroke.

INTRODUCTION

Circadian rhythm disruption (CRD) is a major driver of immune dysregulation; however, whether CRD promotes ischemic stroke (IS) progression through immune imbalance and the underlying molecular mechanisms remain unclear.

METHODS

Transcriptomic data from human IS brain tissues were analyzed to identify CRD associated regulators using machine-learning approaches. A CRD score model was constructed to quantify circadian disruption. Circadian timing-dependent stroke models were established to evaluate diurnal variation in stroke severity and gene rhythmicity in vivo. Single-cell transcriptomic analysis of peripheral blood mononuclear cell -derived scRNA-seq data was performed to characterize CRD-associated immune remodeling. Virtual gene knockdown analysis using the scTenifoldKnk framework, together with experimental validation including microglial gene silencing and T cell co-culture in vitro, as well as adeno-associated virus (AAV)-mediated gene intervention in a transient middle cerebral artery occlusion (tMCAO) model in vivo, were conducted to assess immunoregulatory effects and neurological recovery.

RESULTS

We identified seven hub genes associated with CRD in IS patients. Circadian timing significantly influenced stroke severity, with marked differences in infarct volume across Zeitgeber time points. In parallel, coronin 1C (CORO1C) exhibited rhythmic expression in the brain under physiological conditions, which was disrupted following ischemic stroke. High CRD scores were associated with enhanced CD8+ T-cell activation and reduced M0 macrophages, indicating a proinflammatory immune phenotype. Single-cell analysis revealed activation of PD-1/PD-L1 signaling and Th17 polarization within T-cell subsets, consistent with immune overactivation and functional exhaustion. Virtual knockdown analysis highlighted CORO1C as a central regulator linking neuronal and immune transcriptional programs. In microglia, CORO1C knockdown reduced IL-6 and IFN-γ secretion. In vivo, AAV-shCORO1C treatment decreased cerebral CORO1C expression, attenuated CD8+ T-cell infiltration and inflammatory signaling, enhanced Claudin-5 expression, improved neurological function, and promoted post-stroke recovery.

CONCLUSION

This study indicates that CORO1C acts as a pivotal regulator that mediates circadian rhythm disruption-driven immune dysregulation, thereby influencing stroke progression and neurological recovery.