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USP2 promotes tumor immune evasion via deubiquitination and stabilization of PD-L1

Cell Death & Differentiation volume 30, pages 2249–2264 (2023)Cite this article

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Abstract

The abnormal upregulation of programmed death ligand-1 (PD-L1) on tumor cells impedes T-cell mediated cytotoxicity through PD-1 engagement, and further exploring the mechanisms regulation of PD-L1 in cancers may enhance the clinical efficacy of PD-L1 blockade. Here, using single-guide RNAs (sgRNAs) screening system, we identify ubiquitin-specific processing protease 2 (USP2) as a novel regulator of PD-L1 stabilization for tumor immune evasion. USP2 directly interacts with and increases PD-L1 abundance in colorectal and prostate cancer cells. Our results show that Thr288, Arg292 and Asp293 at USP2 control its binding to PD-L1 through deconjugating the K48-linked polyubiquitination at lysine 270 of PD-L1. Depletion of USP2 causes endoplasmic reticulum (ER)-associated degradation of PD-L1, thus attenuates PD-L1/PD-1 interaction and sensitizes cancer cells to T cell-mediated killing. Meanwhile, USP2 ablation-induced PD-L1 clearance enhances antitumor immunity in mice via increasing CD8+ T cells infiltration and reducing immunosuppressive infiltration of myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs), whereas PD-L1 overexpression reverses the tumor growth suppression by USP2 silencing. USP2-depletion combination with anti-PD-1 also exhibits a synergistic anti-tumor effect. Furthermore, analysis of clinical tissue samples indicates that USP2 is positively associated with PD-L1 expression in cancer. Collectively, our data reveal a crucial role of USP2 for controlling PD-L1 stabilization in tumor cells, and highlight USP2 as a potential therapeutic target for cancer immunotherapy.

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Fig. 1: USP2 positively maintains PD-L1 protein stability.
Fig. 2: USP2 specifically interacts with and stabilizes PD-L1.
Fig. 3: USP2 modulates K48-linked polyubiquitin chains of PD-L1.
Fig. 4: USP2 depletion induced endoplasmic reticulum (ER)-associated degradation of PD-L1.
Fig. 5: Depletion of USP2 promotes the cytotoxicity of T cell toward tumor cell.
Fig. 6: USP2-depletion-mediated PD-L1 degradation promotes antitumor immunity.
Fig. 7: USP2 correlates with PD-L1 abundance in colorectal cancer.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors sincerely thank Prof. Hong-bing Shu for providing HA-Ubiquitin KR plasmids, Prof. Ronggui Hu for providing HA-Ubiquitin K only plasmids, Prof. Han Liu for providing USP2 C276A plasmid, and Prof. Guohui Wan for providing primary human CRC cells. This study was supported by grants from National Natural Science Foundation of China (82273960, 81973366, 82273854, 82003792 and 82304512), CAMS Innovation Fund for Medical Sciences (2021-I2M-1-070), and Beijing Nova Program (20220484116). The funding sources had no involvements in study design, data collection, data analysis, data interpretation, manuscript preparation and submission.

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  1. These authors contributed equally: Zean Kuang, Xiaojia Liu, Na Zhang.

Authors and Affiliations

  1. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100050, China

    Zean Kuang, Na Zhang, Jingwen Dong, Cuicui Sun, Mingxiao Yin, Yuting Wang, Danqing Song & Hongbin Deng

  2. Beijing Institute of Clinical Pharmacy, Beijing Friendship Hospital, Capital Medical University, Beijing, 100050, China

    Xiaojia Liu

  3. Qingdao Women and Children’s Hospital, Qingdao University, Qingdao, 266034, China

    Lu Liu

  4. Beijing Institute of Pharmacology and Toxicology, National Engineering Research Center for the Emergency Drug, Beijing, 100850, China

    Dian Xiao & Xinbo Zhou

  5. National Institutes for Food and Drug Control, Beijing, 102629, China

    Yanchun Feng

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Contributions

HD overall designed, supervised and coordinated the study. ZK and XL performed most of the experiments. NZ, JD, MY, CS, YW and LL participated in the molecular and cellular biological experiments. DX, XZ performed molecular docking. YF and DS provided reagents and performed data analysis. HD supervised the study and interpreted results, wrote and revised the manuscript. All authors read and approved the manuscript.

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Correspondence to Yanchun Feng, Danqing Song or Hongbin Deng.

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Kuang, Z., Liu, X., Zhang, N. et al. USP2 promotes tumor immune evasion via deubiquitination and stabilization of PD-L1. Cell Death Differ 30, 2249–2264 (2023). https://doi.org/10.1038/s41418-023-01219-9

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