HIF-Overexpression and Pro-Inflammatory Priming in Human Mesenchymal Stromal Cells Improves the Healing Properties of Extracellular Vesicles in Experimental Crohn's Disease

Extracellular vesicles (EVs) derived from mesenchymal stromal cells (MSCs) have therapeutic potential in the treatment of several immune disorders, including ulcerative colitis, owing to their regenerative and immunosuppressive properties. We recently showed that MSCs engineered to overexpress hypoxia-inducible factor 1-alpha and telomerase (MSC-T-HIF) and conditioned with pro-inflammatory stimuli release EVs (EVMSC-T-HIF(C)) with potent immunomodulatory activity. We tested the efficacy of EVMSC-T-HIF(C) to repolarize M1 macrophages (M phi 1) to M2-like macrophages (M phi 2-like) by analyzing surface markers and cytokines and performing functional assays in co-culture, including efferocytosis and T-cell proliferation. We also studied the capacity of EVMSC-T-HIF(C) to dampen the inflammatory response of activated endothelium and modulate fibrosis. Finally, we tested the therapeutic capacity of EVMSC-T-HIF(C) in an acute colitis model. EVMSC-T-HIF(C) induced the repolarization of monocytes from M phi 1 to an M phi 2-like phenotype, which was accompanied by reduced inflammatory cytokine release. EVMSC-T-HIF(C)-treated M phi 1 had similar effects of immunosuppression on activated peripheral blood mononuclear cells (PBMC) as M phi 2, and reduced the adhesion of PBMCs to activated endothelium. EVMSC-T-HIF(C) also prevented myofibroblast differentiation of TGF-beta-treated fibroblasts. Finally, administration of EVMSC-T-HIF(C) promoted healing in a TNBS-induced mouse colitis model in terms of preserving colon length and intestinal mucosa architecture and altering the ratio of M phi 1/ M phi 2 infiltration. In conclusion, EVMSC-T-HIF(C) have effective anti-inflammatory properties, making them potential therapeutic agents in cell free-based therapies for the treatment of Crohn's disease and likely other immune-mediated inflammatory diseases.