IL-10 Induction by Extracellular Vesicles from Glycolytic Mesenchymal Stromal Cells: An Innovative Biotherapy for Rheumatoid Arthritis

Mesenchymal stromal cells (MSCs) are multipotent cells with broad immunoregulatory capabilities, positioning them as promising candidates for the treatment of autoimmune diseases. MSC immunoregulatory properties are primarily mediated by the release of paracrine factors including extracellular vesicles (EVs). We have previously demonstrated that the metabolic reprogramming of human umbilical cord MSCs (UC-MSCs) to glycolysis enhances their ability to inhibit proinflammatory T cell proliferation, to reduce the secretion of pro-inflammatory cytokines by T cells and to promote the generation of T regulatory cells (Tregs). In this study, we evaluated and compared the immunoregulatory properties of EVs derived from control and glycolytic UC-MSCs in vitro and in vivo.

EVs were isolated from control and glycolytic UC-MSCs by ultracentrifugation, characterized using nanoparticle tracking analysis (NTA) and flow cytometry (FACS), and assessed for their effects on PBMCs and memory T-CD4 cells isolated from healthy donors. EV internalization was measured on memory T-CD4+ cells by FACS, confocal microscopy and qPCR. In vivo, we investigated their effects in the DTH and the CIA models.

In vitro, we showed that EVs derived from glycolytic MSC induced the generation of Tregs. In vivo, in the DTH model we found that EVs derived from glycolytic MSC decreased proinflammatory T cell infiltration, DTH inflammation compared to EVs derived from control MSC. In the CIA model, EVs derived from glycolytic MSC reduced prevent the development and the progression to a higher rate than EVs isolated from control MSC. This was associated with significantly higher Treg/Th1 and Treg/Th17 ratios in the LN and spleen, respectively, of arthritic mice treated with EVs derived from glycolytic MSCs compared with arthritic mice or mice treated with EVs isolated from control MSCs.

Overall, our findings indicate that EV derived from glycolytic MSC effectively suppress the pro-inflammatory T cell response while promoting the generation of regulatory T cells, highlighting their enhanced therapeutic potential in inflammatory and autoimmune diseases like arthritis.