Integrated analysis of enzymes, mRNAs, and miRNAs provides insight into the regulatory potential of extracellular vesicles in recipient cell glucose metabolism.

INTRODUCTION: Extracellular vesicles (EVs) are increasingly recognized as active coordinators of metabolic processes rather than mere messengers. By carrying unique subsets of enzymes, metabolites, lipids, and nucleic acids, EVs can directly deliver functional metabolic machinery or dynamically alter intracellular metabolic fluxes in recipient cells. However, their role in regulating specific biochemical pathways remains largely unknown. METHODOLOGY: In the current in silico analysis, we explored the dominant metabolic role of EV cargo using publicly available multi-omics data. For this, the top 500 mRNAs and proteins, along with miRNAs reported at least 10 times in humans across independent studies, as catalogued in the EVpedia database are considered and curated into a comprehensive dataset. RESULTS: Enrichment analysis of these mRNAs and proteins revealed that carbohydrate metabolic pathways, including glycolysis, the pentose phosphate pathway and the TCA cycle, were over-represented in EVs. Further, to investigate whether EVs carry miRNAs that regulate these pathways, we analyzed the miRNA targets. Enrichment analysis of EV miRNA targets mapped glycolytic regulatory genes, including HK1, HK2, PFKP and PKM. Interestingly, we also found miRNAs targeting genes encoding glucose transporters (SLC2A1, SLC2A3, SLC2A4, and SLC2A14) reported in EVs. Genomic annotation of these miRNAs revealed them to form clusters, including the miR-17-92 cluster, a well-known regulator of glycolysis. DISCUSSION: While the study has limitations-mainly due to the biological heterogeneity of EVs and the difficulty of standardizing cargo-the results potentially suggest that, by delivering enzymes, their mRNAs, regulatory miRNAs or combinations thereof, EVs could potentially mediate recipient cell glucose metabolism.