Metabolic reprogramming augments potency of human pSTAT3–inhibited iTregs to suppress alloreactivity
Kelly Walton, Mario R. Fernandez, Elizabeth M. Sagatys, Jordan Reff, Jongphil Kim, Marie Catherine Lee, John V. Kiluk, Jane Yuet Ching Hui, David McKenna Jr., Meghan Hupp, Colleen Forster, Michael A. Linden, Nicholas J. Lawrence, Harshani R. Lawrence, Joseph Pidala, Steven Z. Pavletic, Bruce R. Blazar, Said M. Sebti, John L. Cleveland, Claudio Anasetti, Brian C. Betts
Kelly Walton, Mario R. Fernandez, Elizabeth M. Sagatys, Jordan Reff, Jongphil Kim, Marie Catherine Lee, John V. Kiluk, Jane Yuet Ching Hui, David McKenna Jr., Meghan Hupp, Colleen Forster, Michael A. Linden, Nicholas J. Lawrence, Harshani R. Lawrence, Joseph Pidala, Steven Z. Pavletic, Bruce R. Blazar, Said M. Sebti, John L. Cleveland, Claudio Anasetti, Brian C. Betts
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Research Article Immunology Transplantation

Metabolic reprogramming augments potency of human pSTAT3–inhibited iTregs to suppress alloreactivity

Abstract

Immunosuppressive donor Tregs can prevent graft-versus-host disease (GVHD) or solid-organ allograft rejection. We previously demonstrated that inhibiting STAT3 phosphorylation (pSTAT3) augments FOXP3 expression, stabilizing induced Tregs (iTregs). Here we report that human pSTAT3–inhibited iTregs prevent human skin graft rejection and xenogeneic GVHD yet spare donor antileukemia immunity. pSTAT3-inhibited iTregs express increased levels of skin-homing cutaneous lymphocyte-associated antigen, immunosuppressive GARP and PD-1, and IL-9 that supports tolerizing mast cells. Further, pSTAT3-inhibited iTregs significantly reduced alloreactive conventional T cells, Th1, and Th17 cells implicated in GVHD and tissue rejection and impaired infiltration by pathogenic Th2 cells. Mechanistically, pSTAT3 inhibition of iTregs provoked a shift in metabolism from oxidative phosphorylation (OxPhos) to glycolysis and reduced electron transport chain activity. Strikingly, cotreatment with coenzyme Q10 restored OxPhos in pSTAT3-inhibited iTregs and augmented their suppressive potency. These findings support the rationale for clinically testing the safety and efficacy of metabolically tuned, human pSTAT3–inhibited iTregs to control alloreactive T cells.

Authors

Kelly Walton, Mario R. Fernandez, Elizabeth M. Sagatys, Jordan Reff, Jongphil Kim, Marie Catherine Lee, John V. Kiluk, Jane Yuet Ching Hui, David McKenna Jr., Meghan Hupp, Colleen Forster, Michael A. Linden, Nicholas J. Lawrence, Harshani R. Lawrence, Joseph Pidala, Steven Z. Pavletic, Bruce R. Blazar, Said M. Sebti, John L. Cleveland, Claudio Anasetti, Brian C. Betts

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Figure 8

Human pSTAT3–inhibited iTregs maintain antileukemia immunity by donor T cells.

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Human pSTAT3–inhibited iTregs maintain antileukemia immunity by donor T ...
Graph depicts mean lysis ± SEM by U937-specific, human T cells generated in vivo using NSG mice transplanted with human PBMCs, pSTAT3-inhibited (S3i) or control iTregs, and then vaccinated with irradiated U937cells (2 × 106, ATCC) on days 0 and +7. Human T cells from nonvaccinated mice containing PBMCs alone group served as a negative control. On day +12, the mice were euthanized, and human T cells were harvested and purified from the spleen using magnetic bead separation. Tregs were specifically not removed from the harvested T cells. The purified T cells were cultured with fresh U937 targets at varying ratios. U937 lysis was measured using a colorimetric assay after 4 hours of culture. Results shown are from 1 of 2 independent experiments, using 4 mice per group. ANOVA. ****P < 0.0001. pSTAT3, STAT3 phosphorylation; iTregs, induced Tregs.