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 5

Human pSTAT3–inhibited iTregs significantly reduce pathogenic Th1 cells in the spleen.

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Human pSTAT3–inhibited iTregs significantly reduce pathogenic Th1 cells ...
NSG mice received a human skin graft, allogeneic PBMCs, and pSTAT3-inhibited (S3i) or control iTregs. On day +21, the recipient spleens were harvested and analyzed for human T cell numbers and effector subsets. Graphs show the absolute numbers of (A) CD4+ and (B) CD8+ central memory (CD62L+, CD45RO+), effector memory (CD62L–, CD45RO+), and naive T cells (CD62L+, CD45RO–) ± SEM. (C–G) Box and-whisker plots (max, min, median) show the frequency and absolute number of human Th1 (CD4+, IFN-γ+) and Th2 cells (CD4+, IL-4+), with representative contour plots showing the T cell populations. n = 3 experiments, up to 11 mice/group. Human pSTAT3–inhibited or DMSO-treated iTregs were cultures with autologous CD4+ T cells with allogeneic DCs (T cell to DC ratio 30:1) for 5 days. The frequency of skin-homing CLA expression (H) on the CD4+, IL-4+ (max, min, median) was determined by flow cytometry. (I) Representative contour plots are shown. n = 4 independent experiments. ANOVA (A–D, F, and G) or paired t test (H). *P < 0.05 or **P = 0.001–0.01. pSTAT3, STAT3 phosphorylation; iTregs, induced Tregs.