Glycometabolic regulation of the biogenesis of small extracellular vesicles

The biogenesis of small extracellular vesicles (sEVs) is regulated by multiple molecular machineries, generating considerably heterogeneous vesicle populations, i.e., exosomes and non-exosomal vesicles, with distinct cargo molecules. However, the role of carbohydrate metabolism in generating such vesicle heterogeneity remains largely elusive. Here we discovered that 2-deoxyglucose (2-DG), a well-known glycolysis inhibitor, suppressed the secretion of non-exosomal vesicles by impairing asparagine-linked glycosylation (N-glycosylation) in mouse melanoma cells. In the present study, to gain deeper insight into the molecular signatures of non-exosomal vesicles, we performed label-free quantitative proteomics of sEVs released from cells that were treated with or without 2-DG or NGI-1.