Donor T cell STAT3 deficiency enables tissue PD-L1–dependent prevention of graft-versus-host disease while preserving graft-versus-leukemia activity
Qinjian Li, … , Xi Zhang, Defu Zeng
Qinjian Li, … , Xi Zhang, Defu Zeng
Published August 1, 2023
Citation Information: J Clin Invest. 2023;133(15):e165723. https://doi.org/10.1172/JCI165723.
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Research Article Hematology

Donor T cell STAT3 deficiency enables tissue PD-L1–dependent prevention of graft-versus-host disease while preserving graft-versus-leukemia activity

Abstract

STAT3 deficiency (STAT3–/–) in donor T cells prevents graft-versus-host disease (GVHD), but the impact on graft-versus-leukemia (GVL) activity and mechanisms of GVHD prevention remains unclear. Here, using murine models of GVHD, we show that STAT3–/– donor T cells induced only mild reversible acute GVHD while preserving GVL effects against nonsusceptible acute lymphoblastic leukemia (ALL) cells in a donor T cell dose–dependent manner. GVHD prevention depended on programmed death ligand 1/programmed cell death protein 1 (PD-L1/PD-1) signaling. In GVHD target tissues, STAT3 deficiency amplified PD-L1/PD-1 inhibition of glutathione (GSH)/Myc pathways that regulate metabolic reprogramming in activated T cells, with decreased glycolytic and mitochondrial ATP production and increased mitochondrial ROS production and dysfunction, leading to tissue-specific deletion of host-reactive T cells and prevention of GVHD. Mitochondrial STAT3 deficiency alone did not reduce GSH expression or prevent GVHD. In lymphoid tissues, the lack of host-tissue PD-L1 interaction with PD-1 reduced the inhibition of the GSH/Myc pathway despite reduced GSH production caused by STAT3 deficiency and allowed donor T cell functions that mediate GVL activity. Therefore, STAT3 deficiency in donor T cells augments PD-1 signaling–mediated inhibition of GSH/Myc pathways and augments dysfunction of T cells in GVHD target tissues while sparing T cells in lymphoid tissues, leading to prevention of GVHD while preserving GVL effects.

Authors

Qinjian Li, Xiaoqi Wang, Qingxiao Song, Shijie Yang, Xiwei Wu, Dongyun Yang, Isabelle J. Marié, Hanjun Qin, Moqian Zheng, Ubaydah Nasri, Xiaohui Kong, Bixin Wang, Elizabeth Lizhar, Kaniel Cassady, Josh Tompkins, David Levy, Paul J. Martin, Xi Zhang, Defu Zeng

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Figure 6

Stat3 deficiency in donor T cells augments PD-1–mediated inhibition of GSH/Myc pathways and production of Mito-ROS.

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Stat3 deficiency in donor T cells augments PD-1–mediated inhibition of G...
Lethally irradiated BALB/c recipients were engrafted with TCD-BM and CD90.2+ T cells from WT, STAT3–/–, or STAT3–/–PD-1–/– C57BL/6 donors as described for Figure 5. On day 6 after HCT, CD90.2+ T cells from liver were isolated for RNA-Seq, Seahorse, immunoblotting, and flow-cytometry analysis. Data are combined from at least 2 replicate experiments. (A) NES of KEGG pathway activity of CD4+ T cells, setting the activity of WT CD4+ T cells as the reference, for comparisons with STAT3–/– and STAT3–/–PD-1–/– T cells. (B) GSEA plots of MYC target V2 pathway-related gene set expression in WT, STAT3–/–, and STAT3–/–PD-1–/– donor CD4+ T cells. (C) Myc protein was measured by immunoblotting. (D) Glyco-ATP, Mito-ATP, and total ATP. n = 7–14. (E) The GSH metabolism pathway is shown. (F and G) MFI of CD98 and reduced GSH of CD4+ T cells in the liver. n = 5–9. (H) Glycolysis pathway is shown. (I and J) MFI of GLUT1 and HK2 of CD4+ T cells. n = 5–9. (K) FAO pathway is shown. (L and M) MFI of CD36 and CPT1A of CD4+ T cells. n = 5–9. (N) MFI of Trx1 of CD4+ T cells. n = 5–9. (O) Representative flow cytometry pattern and mean ± SEM of percentage MitoSOXhiMitoGreenhi and percentage MitoRedloMitoGreenhi CD4+ T cells. n = 5–9. Separate experiments were performed with WT versus STAT3–/– or STAT3–/– versus STAT3–/–PD-1–/– in D, F, G, I, J, L, M, N, and results were normalized to the mean values for STAT-3–deficient cells. Data are represented as mean ± SEM. P values were calculated by using 1-way ANOVA (C, D, F, G, I, J, L, and O). NS, P ≥ 0.05; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.