Session: 701. Experimental Transplantation: Basic and Translational: Poster III
Hematology Disease Topics & Pathways:
Fundamental Science, Research, Translational Research, GVHD, Diseases, Immune Disorders
To address the question, we initially analyzed the publicly available RNA sequencing (RNA seq) data and found that Eif2ak3 (encoding PERK) were dramatically increased in allogenic T cells. We next established syngeneic and allogeneic murine models of HCT and observed that PERK and ERAD-associated genes were significantly upregulated in donor T cells transferred into allogeneic recipients as compared with those of syngeneic controls. Upon allogenic stimulation, T cells increased phosphorylation of PERK (p-PERK). To further assess the role of PERK in T cell-driven GVHD, we purified T cells from WT and PERKflox/floxCD4Cre (PERK-cKO) mice and transferred into lethally irradiated BALB/c mice and proved that PERK deficiency in donor T cells significantly attenuated GVHD while preserving GVL effect. The outcome was associated with reduced proliferation of CD4+ but not CD8+ T cells and Th1 and Th17 differentiation, and increased Treg generation via downregulating Nrf2 levels in PERK-deficient T cells.
Mechanistically, we found that PERK interacted with SEL1L in activated T cells. Interestingly, PERK differentially regulated the pro-inflammatory cytokines secretion in T cells upon allogenic versus anti-CD3 polyclonal stimulation. PERK increased the generation of Th1 and Th17 and augmented GVHD while downregulating SEL1L levels. In contrast, PERK inhibited the pro-inflammatory cytokine production by T cells after anti-CD3 stimulation while upregulating SEL1L levels. Furthermore, we demonstrated that PERK enhanced mitochondrial membrane potential (MMP) in CD4+ T cells upon allogeneic stimulation through SEL1L-mediated ERAD pathway, thus decreasing the protein levels of PINK1 and inhibiting the autophagy of allogenic T cells.
Pharmacologically, treatment of allo-HCT recipients with PERK specific inhibitor, AMG44, significantly reduced GVHD severity while maintaining the GVL activity. For translational research, we also assessed the effect of AMG44 in GVHD induced by human PBMCs in NSG HLA-A2-transgenic mice. We found that AMG44 administration significantly reduced GVHD severity reflected by prolonged survival, reduced GVHD scores and body weight loss. More importantly, the treatment with AMG44 preserved the GVL effect against B-cell lymphoma.
In summary, our findings validate PERK as a potential novel therapeutic target for the prevention of GVHD while preserving GVL responses. The study elucidates a new mechanism by which PERK differentially regulated T-cell allogeneic and polyclonal responses through modulating ERAD pathway.
Disclosures: No relevant conflicts of interest to declare.
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