Abstract

Mesenchymal stromal cells (MSCs) exert potent immunomodulatory effects largely mediated by extracellular vesicles (EVs). We previously demonstrated that glycolytic reprogramming enhances the immunosuppressive capacity of human umbilical cord-derived MSCs (UC-MSCs). Here, we investigated whether this enhanced activity is transmitted through EVs and explored the contribution of EV-associated microRNAs. Methods: EVs from na & iuml;ve and glycolytically reprogrammed UC-MSCs (EVs-UC-MSCnaive and EVs-UC-MSCglyco) were isolated, characterized, and tested for their effects on memory CD4(+)T and B cells in vitro and their therapeutic efficacy in vivo in the delayed-type hypersensitivity (DTH) and in the collagen induced arthrtis (CIA) murine model. Results: EVs-UC-MSCglyco more effectively suppressed inflammatory T cell responses, promoted IL-10-producing Tr1 and B cells, and enhanced B cell survival compared with EVs-UC-MSCnaive. In vivo, EVs-UC-MSCglyco significantly reduced inflammation in a DTH murine model and decreased arthritis incidence and clinical severity in CIA. These effects were associated with increased Treg/Th1, Treg/Th17, Tr1/Th1, and Tr1/Th17 ratios, together with enhanced IL-10 production. MicroRNA profiling revealed enrichment of regulatory miRNAs, including miR-365a-5p, linked to suppression of pro-inflammatory signaling and activation of the IL-10 regulatory axis. Conclusions: Glycolytic reprogramming enhances the therapeutic potential of UC-MSC-derived EVs, highlighting EVs-UC-MSCglyco as promising immunomodulatory candidates for the treatment of autoimmune diseases such as arthritis.