Extracellular Vesicle Phenotypic Trajectories During Alcohol Withdrawal Treatment

Alcohol dependence or alcohol use disorder is a chronic mental disorder that imposes a heavy global burden. Small extracellular vesicles (EVs) are emerging as a promising source of real-time information about brain status, yet existing studies are predominantly cross-sectional. We aimed to characterize longitudinal changes in EV properties and immunophenotype during alcohol withdrawal treatment. In our study plasma-derived EVs were analyzed and immunophenotyping via flow cytometry was performed to measure the concentrations of vesicle fractions carrying erythrocyte-, platelet-, endothelial-, and neuronal-associated markers in patients undergoing alcohol withdrawal and in healthy volunteers. Patient-derived EVs exhibited greater size heterogeneity, and microscopy images revealed vesicles with altered morphology. Concentrations of EVs bearing NLGN3 were lower in patients. CD62P+ and CD146+ EVs were also less represented in patients. Notably, CD31+ EVs separated into two distinct clusters; their concentrations were higher in patients with a binge drinking pattern but markedly lower in those with a steady pattern. The findings reveal dynamic phenotypic shifts in circulating EVs during alcohol withdrawal. The divergent patterns according to alcohol use history highlight the potential of EV profiling for personalized monitoring and underscore the necessity of longitudinal designs in vesicle-based biomarker research.