Role of miR-449a in the Activation and Metabolism of CD4⁺ T Cells

Background: Acute rejection is a significant challenge after organ transplantation. The CD4⁺ T-cell‒mediated immune response plays an important role in acute transplant rejection. It was also found that miR-449a microRNA regulates the alloimmune response in a model of heart transplantation in mice. Our goal was to determine the role of miR-449a in the regulation of CD4⁺ T cells.

Methods: We examined miR-449a expression in peripheral blood mononuclear cells (PBMCs) and graft-infiltrating lymphocytes (GILs) between syngeneic transplant and allogeneic transplant groups on day 7 post‒heart transplantation. We also examined miR-449a expression in CD4⁺ T-cell activation and mixed-lymphocyte reactions (MLRs) in vitro. To evaluate the effect of miR-449a on CD4⁺ T-cell metabolism, we analyzed key metabolic parameters using XFp extracellular flux analyses.

Results: Our in vivo heart transplant models showed that the expression of miR-449a in PBMCs and in GILs significantly increased in the allogeneic groups in comparison to the syngeneic groups (P < .01). Furthermore, in vitro analysis confirmed that the expression of miR-449a was significantly elevated in activated CD4⁺ T cells. Reduction of miR-449a expression in CD4⁺ T cells decreased the mitochondrial respiration in the same CD4⁺ T cells.

Conclusion: Our results reveal that miR-449a microRNA was elevated in allogeneic heart allografts. This correlated with an increased miR-449a expression in activated CD4⁺ T cells. Inhibition of miR-449a in activated CD4⁺ T cells coincided with reduced mitochondrial respiration, suggesting that miR-449a influences CD4⁺ T-cell activation during the alloimmune response by regulating metabolic status.