ER stress sensor PERK promotes T cell pathogenicity in GVHD by regulating ER-associated degradation
Qiao Cheng, Hee-Jin Choi, Yongxia Wu, Xiaohong Yuan, Allison Pugel, Linlu Tian, Michael Hendrix, Denggang Fu, Reza Alimohammadi, Chen Liu, Xue-Zhong Yu
Qiao Cheng, Hee-Jin Choi, Yongxia Wu, Xiaohong Yuan, Allison Pugel, Linlu Tian, Michael Hendrix, Denggang Fu, Reza Alimohammadi, Chen Liu, Xue-Zhong Yu
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Research Article Immunology Inflammation

ER stress sensor PERK promotes T cell pathogenicity in GVHD by regulating ER-associated degradation

Abstract

Endoplasmic reticulum (ER) stress through IRE1/XBP1 is implicated in the onset and progression of graft-versus-host disease (GVHD), but the role of the ER stress sensor PERK in T cell allogeneic responses and GVHD remains unexplored. Here, we report that PERK is a key regulator in T cell allogeneic response and GVHD induction. PERK augments GVHD through increasing Th1 and Th17 population, while reducing Treg differentiation by activating the Nrf2 pathway. Genetic deletion or selective inhibition of PERK pharmacologically reduces GVHD while preserving graft-versus-leukemia (GVL) activity. At the cellular level, PERK positively regulates CD4+ T cell pathogenicity while negatively regulating CD8+ T cell pathogenicity in the induction of GVHD. At the molecular level, PERK interacts with SEL1L and regulates SEL1L expression, leading to augmented T cell allogeneic responses and GVHD development. In vivo, PERK deficiency in donor T cells alleviates GVHD through ER-associated degradation. Furthermore, pharmacological inhibition of PERK with AMG44 significantly suppresses the severity of GVHD induced by murine or human T cells. In summary, our findings validate PERK as a potential therapeutic target for the prevention of GVHD while preserving GVL responses, and uncover the mechanism by which PERK differentially regulates CD4+ versus CD8+ T cell allogeneic and antitumor responses.

Authors

Qiao Cheng, Hee-Jin Choi, Yongxia Wu, Xiaohong Yuan, Allison Pugel, Linlu Tian, Michael Hendrix, Denggang Fu, Reza Alimohammadi, Chen Liu, Xue-Zhong Yu

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Figure 6

PERK regulates T cell allogeneic responses through ERAD.

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PERK regulates T cell allogeneic responses through ERAD. (A) T cells fro...
(A) T cells from WT or PERK-cKO mice stimulated with anti-CD3/CD28 (2 μg/mL) or allogeneic APCs for 4 days were performed RNA-Seq. Differentially expressed ERAD-associated genes are shown. (B and C) T cells from WT or PERK-cKO mice were stimulated by allogeneic APCs; mRNA levels of indicated were analyzed by quantitative real-time PCR (B). (C) Protein levels of SEL1L, ERLEC1, and GAPDH were evaluated by Western blot. (D–F) Lethally irradiated BALB/c mice received TCD-BM cells (5 × 106) alone or together with T cells (1.25 × 106) from WT, PERK-cKO, SEL1L-cKO, or PERK/SEL1L-double-KO (dKO) donors. Survival (D), body weight (E), and GVHD scores (F) of BALB/c recipients were monitored after BMT; n = 9–10 combined from 2 replicate experiments. (G–K) Lethally irradiated BALB/c mice received BM cells (5 × 106) isolated from Rag1-KO mice along with T cells (1.25 × 106) purified from WT, PERK-cKO, SEL1L-cKO, or PERK/SEL1L-dKO donors. (G) Levels of IFN-γ, TNF-α, and GM-CSF in donor CD4+ or CD8+ T cells of livers from the recipients were analyzed. (H) Percentages of IFN-γ+CD4+, TNF-α+CD4+, and GM-CSF+CD4+ T cells among gated H2Kb+CD4+ T cells are shown. (I) Percentages of IFN-γ+CD8+, TNF-α+CD8+, and GM-CSF+CD8+ T cells among gated H2Kb+CD8+ T cells are shown. (J) Levels of CHOP in donor CD4+ T cells were analyzed. (K) Mean fluorescence intensity of CHOP among gated H2Kb+CD4+ T cells is displayed. Log-rank (Mantel-Cox) test (D) and non-parametric Mann-Whitney U test (E and F) were used to compare groups. Data in B are represented as mean ± SD; significance was determined using a 2-tailed unpaired Student’s t test. Data in H, I, and K are represented as mean ± SD; significance was determined using a 1-way ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.