The ATP synthase inhibition induces an AMPK-dependent glycolytic switch of mesenchymal stem cells that enhances their immunotherapeutic potential

Abstract

Objectives: Mesenchymal Stem/Stromal Cells (MSC) are promising therapeutic tools for inflammatory diseases due to their potent immunoregulatory capacities. Their suppressive activity mainly depends on inflammatory cues that have been recently associated with changes in MSC bioenergetic status towards a glycolytic metabolism. However, the molecular mechanisms behind this metabolic reprogramming and its impact on MSC therapeutic properties have not been investigated. Methods: Human and murine-derived MSC were metabolically reprogramed using pro-inflammatory cytokines, an inhibitor of ATP synthase (oligomycin), or 2-deoxy-D-glucose (2DG). The immunosuppressive activity of these cells was tested in vitro using co-culture experiments with pro-inflammatory T cells and in vivo with the Delayed-Type Hypersensitivity (DTH) and the Graph versus Host Disease (GVHD) murine models. Results: We found that the oligomycin-mediated pro-glycolytic switch of MSC significantly enhanced their immunosuppressive properties in vitro . Conversely, glycolysis inhibition using 2DG significantly reduced MSC immunoregulatory effects. Moreover, in vivo , MSC glycolytic reprogramming significantly increased their therapeutic benefit in the DTH and GVHD mouse models. Finally, we demonstrated that the MSC glycolytic switch effect partly depends on the activation of the AMPK signaling pathway. Conclusion: Altogether, our findings show that AMPK-dependent glycolytic reprogramming of MSC using an ATP synthase inhibitor contributes to their immunosuppressive and therapeutic functions, and suggest that pro-glycolytic drugs might be used to improve MSC-based therapy.

Potential mechanisms and therapeutic targets of mesenchymal stem cell transplantation for ischemic stroke

“…3D scaffolds could promote MSCs expansion and increase anti-inflammatory properties [ 40 , 93 ]. Oligomycin or pro-inflammatory cytokines increased the immunosuppressive properties of MSCs by activating AMPK signaling [ 94 ]. Metabolic reprogramming of GMP-grade hUCMSCs increased their suppressive potential in acute graft vs. host disease [ 95 ].…”

Modulation of Inherent Niches in 3D Multicellular MSC Spheroids Reconfigures Metabolism and Enhances Therapeutic Potential

“…Alternatively, the constructed MSC spheroids were enzymatically dissociated using trypsin, and the viability of the cells in the cell suspensions was determined using a trypan blue dye exclusion assay. For 2-deoxyglucose (2-DG) treatment, cell spheroids were treated with 5 mM 2-DG (Sigma-Aldrich, St. Louis, MO, USA) for 24 h prior to analysis [16].…”

The regulatory networks of the Hippo signaling pathway in cancer development

“…Oxidative stress is the state of imbalance between oxidation and antioxidant when cells need antioxidants to neutralize excessive free radicals 51 . Protein kinase AMP-activated catalytic subunit alpha 1 (AMPK) is a crucial regulator of bioenergy metabolism, and the increase of adenine phosphoribosyl transferase (AMP)/ATP content in the body could activate AMPK 52 . Under hypoxic conditions, the content of ATP decreases, and the cells are in an energy-deficient state.…”