Abstract
Upregulation of programmed death ligand 1 (PD-L1) helps tumor cells escape from immune surveillance, and therapeutic antibodies targeting PD-1/PD-L1 have shown better patient outcomes only in several types of malignancies. Recent studies suggest that the clinical efficacy of anti-PD-1/PD-L1 treatments is associated with PD-L1 levels; however, the underlying mechanism of high PD-L1 protein levels in cancers is not well defined. Here, we report that the deubiquitinase OTUB1 positively regulates PD-L1 stability and mediates cancer immune responses through the PD-1/PD-L1 axis. Mechanistically, we demonstrate that OTUB1 interacts with and removes K48-linked ubiquitin chains from the PD-L1 intracellular domain in a manner dependent on its deubiquitinase activity to hinder the degradation of PD-L1 through the ERAD pathway. Functionally, depletion of OTUB1 markedly decreases PD-L1 abundance, reduces PD-1 protein binding to the tumor cell surface, and causes increased tumor cell sensitivity to human peripheral blood mononuclear cells (PBMCs)-mediated cytotoxicity. Meanwhile, OTUB1 ablation-induced PD-L1 destabilization facilitates more CD8+ T cells infiltration and increases the level of IFN-γ in serum to enhance antitumor immunity in mice, and the tumor growth suppression by OTUB1 silencing could be reversed by PD-L1 overexpression. Furthermore, we observe a significant correlation between PD-L1 abundance and OTUB1 expression in human breast carcinoma. Our study reveals OTUB1 as a deubiquitinating enzyme that influences cancer immunosuppression via regulation of PD-L1 stability and may be a potential therapeutic target for cancer immunotherapy.
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Acknowledgements
The authors sincerely thank Prof. Lingqiang Zhang for providing human deubiquitinase OTU subfamily plasmids and several shDUBs vectors, Profs. Jiadong Wang and Ceshi Chen for providing 4T1 cells, and Prof. Wensheng Wei for providing the CRISPR/Cas9 related plasmids. We thank Dr. Weicheng Zang for helpful discussion, and the National Center for Protein Sciences at Peking University, particularly Liying Du and Hongxia Lv for technical help. We also appreciate the assistance of Xiaochen Li from the Core Facilities of Life Sciences at Peking University for their assistance with microscopic imaging. We thank the Human Genetic Resource Core Facility at Peking University for providing peripheral blood samples from Peking University Hospital. This work was supported by the National Key Research and Development Program of China (2016YFC1302401) and the National Natural Science Foundation of China (81730080, 31670786, 32000917).
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D.Z. designed and performed the experiments, analyzed the data, and wrote the manuscript. R.X., X.H., and J.Z. performed the experiments. Z.T. performed the bioinformatics analysis. Y.T. performed the cancer cell injection into mice. X.Z. supervised this study and wrote the manuscript.
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All experiments using human samples and mice were approved by the Ethics Committee of Peking University and complied with all relevant ethical guidelines.
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Zhu, D., Xu, R., Huang, X. et al. Deubiquitinating enzyme OTUB1 promotes cancer cell immunosuppression via preventing ER-associated degradation of immune checkpoint protein PD-L1. Cell Death Differ 28, 1773–1789 (2021). https://doi.org/10.1038/s41418-020-00700-z
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DOI: https://doi.org/10.1038/s41418-020-00700-z
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