Sleep disruption exacerbates allergic airway inflammation through sympathetic-mediated activation of the mtDNA-cGAS-STING pathway in pulmonary macrophages

Sleep disruption (SD) is increasingly recognized as a systemic stressor that worsens inflammatory disease, yet how it reshapes lung immunity remains poorly defined. To address this, we combined an ovalbumin (OVA)-induced allergic airway inflammation model with sleep disruption paradigms, together with transcriptomic profiling, in vivo pharmacological interventions, and in vitro RAW264.7 macrophage-like cells. In an OVA-induced allergic airway inflammation model, SD markedly increased methacholine responsiveness, aggravated peribronchial inflammation and mucus metaplasia, and promoted a more severe inflammatory airway phenotype. These changes were accompanied by enhanced pulmonary catecholaminergic signaling, reflected by increased tyrosine hydroxylase immunoreactivity and elevated norepinephrine levels. Mechanistically, SD induced mitochondrial dysfunction in the lung, characterized by ultrastructural injury, increased mitochondrial reactive oxygen species, and accumulation of cytosolic mtDNA, together with activation of the cGAS-STING pathway. Cell-type localization analysis showed that activated STING signaling was predominantly associated with F4/80-positive pulmonary macrophages, whereas overlap with Ly6G-positive neutrophils was limited. In vivo, chemical sympathectomy with 6-hydroxydopamine attenuated airway hyperresponsiveness and reduced pulmonary STING pathway activation. In RAW 264.7 cells, norepinephrine (NE) amplified lipopolysaccharide (LPS)-induced inflammatory responses, promoted mitochondrial oxidative stress, increased cytosolic mtDNA accumulation, and enhanced IRF3 activation, effects that were attenuated by adrenergic blockade and STING inhibition. Together, these findings support a model in which sleep disruption increases catecholaminergic exposure in allergic lungs, thereby driving macrophage mitochondrial stress and mtDNA-cGAS-STING dependent inflammatory amplification. This work identifies a brain-to-lung neuroimmune mechanism linking sleep loss to worsened airway disease and highlights sympathetic-macrophage signaling as a potential mechanism-informed therapeutic target. Sleep disruption enhances sympathetic nervous system activity, leading to increased norepinephrine (NE) release to the lung. NE acts on pulmonary macrophages, likely through adrenergic signaling pathways, inducing mitochondrial oxidative stress and structural disruption. This is characterized by elevated mitochondrial ROS, which promotes the release and cytosolic accumulation of mitochondrial DNA (mtDNA). The released mtDNA activates the cGAS-STING signaling pathway, leading to TBK1-IRF3 activation and downstream inflammatory gene expression, ultimately contributing to enhanced airway inflammation, mucus hypersecretion, and airway hyperresponsiveness under allergen challenge. Created in BioRender. Haotian, C. (2026) https://BioRender.com/xywyr61.