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STEM CELL TRANSPLANTATION

Thrombin receptor activating peptide-6 decreases acute graft-versus-host disease through activating GPR15

Abstract

G-protein coupled receptor 15 (GPR15) is expressed on T-cells. We previously reported knockout of GPR15 increased acute graft-versus-host disease (GvHD) in mice. In this study, we identified thrombin receptor activating peptide-6 (TRAP-6, peptide sequence: SFLLRN) as an activator of GPR15. GRP15 and β-arrestin2 were needed for TRAP-6-mediated inhibition of mixed lymphocyte reactions. TRAP-6 decreased acute GvHD in allotransplant models in mice, an effect dependent on GPR15-expression in donor T-cells. RNA-seq and protein analyses indicated TRAP-6 increased binding of β-arrestin2/TAB1 and inhibited phosphorylation of TAK1 and NF-κB-P65. GPR15 is expressed differently on CD4+ T-cells and CD8+ T-cells. TRAP-6 inhibited phosphorylation of NF-κB-P65 in CD4+ T-cells but increased granzyme B expression in CD8+ T-cells. TRAP-6 decreased acute GvHD without inhibiting graft-versus-tumor (GvT) efficacy against A20 lymphoma cells. SALLRN, a mutant of TRAP-6, preserved the anti-acute GvHD effect but avoided the adverse effects of TRAP-6. TRAP-6 and SALLRN also decreased allogeneic and xenogeneic reactions induced by human blood mononuclear cells. In conclusion, TRAP-6 activated GPR15 on T-cells and decreased acute GvHD in mice without impairing GvT efficacy.

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Fig. 1: TRAP-6 activates GPR15/β-arrestin2 signal and inhibits mixed lymphocyte reaction in vitro.
Fig. 2: TRAP-6 decreases murine acute GvHD in a GPR15-dependent manner.
Fig. 3: The TRAP-6/GPR15 interaction and its impact on T-cell function in vitro.
Fig. 4: TRAP-6 decreases activation of NF-κB.
Fig. 5: TRAP-6 mutant (SALLRN) does not induce platelet aggregation but preserves the anti-alloreaction effect.
Fig. 6: TRAP-6 and Mutant #2 do not impair GvT effect.
Fig. 7: TRAP-6 and Mutant #2 inhibited allogeneic/xenogeneic reaction induced by human BMCs.

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Data availability

The datasets generated during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

Prof. Robert Peter Gale (Imperial College of Science, Technology and Medicine) kindly revised the typescript.

Funding

This study is supported by National Natural Science Foundation of China (82170214, 82370218 and 81970159 to BP, 81871263 and 81930005 to KX), the Jiangsu Provincial Key Research and Development Program (BE2021631 to BP) and the Natural Science Research of the Jiangsu Higher Education Institutions of China (20KJA320002 to BP). This study also received funding from the Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX22-2915).

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BP designed the project, analyzed data and revised the manuscript. KX and TI contributed to concept, revised the manuscript, and helped to design experiments. CL performed experiments, analyzed data and wrote the manuscript. QL and SC performed experiments and analyzed data (animal models, protein assays, and platelet detection). FZ, YL, JS and YW performed part of experiments. CL, QL and SC contributed equally to this study.

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Correspondence to Takayuki Ikezoe, Kailin Xu or Bin Pan.

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Liu, C., Lan, Q., Cao, S. et al. Thrombin receptor activating peptide-6 decreases acute graft-versus-host disease through activating GPR15. Leukemia 38, 1390–1402 (2024). https://doi.org/10.1038/s41375-024-02212-y

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