Neutrophil-derived extracellular vesicles in the plasma of alpha-1 antitrypsin deficient individuals reveal pro-inflammatory metabolic and transcriptomic signatures

Abstract

Alpha-1 antitrypsin deficiency (AATD) is a genetic disorder caused by mutations in SERPINA1, leading to chronic obstructive pulmonary disease (COPD) and liver disease. Neutrophils are key regulators of inflammatory signaling networks; however, their dysregulation in AATD and the underlying molecular mechanisms remain poorly understood. Here, we employed a multi-omics approach integrating RNA sequencing (RNA-seq) and metabolomics to comprehensively characterize neutrophil dysfunction in AATD. RNA-seq analysis of blood neutrophils from AATD individuals revealed transcriptional dysregulation in genes involved in intracellular signaling, immune response regulation, and metabolic adaptation. Isolation and characterization of neutrophil-derived extracellular vesicles (EV) demonstrated an increased plasma burden of neutrophil elastase (NE)-rich EV with elevated surface-bound NE. Metabolomic profiling revealed that these EVs are enriched with pro-inflammatory metabolites linked to dysregulated signaling pathways. Integrated transcriptomic and metabolomic network analysis showed that altered neutrophil gene expression and signaling pathways reshape EV metabolic cargo, linking metabolic reprogramming to inflammatory signal transduction in AATD. Furthermore, differentially expressed EV metabolites may modulate gene expression in recipient cells, sustaining chronic inflammation in AATD. The observed upregulation of interferon, pattern recognition receptors, and cytokine-mediated signaling pathways in neutrophils suggests a potential feedback loop amplifying inflammation in AATD and COPD.