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Destruction of VISTA by TRIM25 ablation in T cells potentiates cancer immunotherapy

Cell Research volume 35, pages 1003–1020 (2025)Cite this article

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Abstract

The limited success of current immunotherapies emphasizes the need for new targets and combination treatments. V-domain Ig suppressor of T cell activation (VISTA) is a promising immune checkpoint target in cancer immunotherapy, but its regulatory mechanism is poorly understood. Through CRISPR knockout screening and proteomic analysis, we identify tripartite motif containing 25 (TRIM25) as a positive regulator for VISTA largely through antagonizing its degradation signaling. Moreover, ERK-mediated phosphorylation of VISTA at Thr284 enhances its interaction with TRIM25, leading to VISTA stabilization. A VISTA-derived phospho-peptide competitively disrupts TRIM25–VISTA interaction, thereby reducing VISTA expression and potentiating the anti-tumor efficacy of PD-1/PD-L1 blockade. Moreover, single-cell RNA sequencing analysis shows that tumor-infiltrating cytotoxic CD8+ T cells are increased in mice with T cell-specific knockout of Trim25. Of note, genetic ablation of Trim25 in T cells not only improves anti-PD-L1 immunotherapy, but also significantly ameliorates CAR T anti-tumor activity in various mouse tumor models. Collectively, this study unveils a mechanism for VISTA regulation in T cells and highlights targeting TRIM25–VISTA as a potential strategy to enhance tumor immunotherapy.

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Fig. 1: CRISPR KO screen and proteomic analysis identify TRIM25 as a key regulator for VISTA protein homeostasis.
Fig. 2: TRIM25 interacts with VISTA and promotes its K63-linked ubiquitination.
Fig. 3: 14-3-3ε recognizes ERK-mediated phosphorylation of VISTA at Thr284, facilitating the interaction between VISTA and TRIM25.
Fig. 4: Knockout of Trim25 decreases VISTA in T cells and enhances anti-tumor immunity.
Fig. 5: Trim25 cKO mice exhibit improved tumor control in the autochthonous tumor model.
Fig. 6: Trim25 deficiency in T cells enhances the efficacy of immune checkpoint blockade.
Fig. 7: Trim25 KO CAR T cells display enhanced anti-tumor efficacy.

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Data availability

All data are available in the main text or the supplementary materials. The raw DNA sequencing data of the CRISPR screen have been deposited under SRA PRJNA1183302. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the iProX partner repository with the dataset identifier PXD057240. The scRNA-seq FASTQ files are available at GEO under accession GSE281314. All the materials generated in this study are available upon reasonable request to the lead contact.

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Acknowledgements

We thank the Core Facility and the Animal Facility of Medical Research Institute of Wuhan University for technical support. This work was supported by grants from the National Key R&D Program of China (2022YFC3401500 and 2023YFC3402100 to J.Z.), the National Natural Science Foundation of China (82273062 to J.Z., 82103149 to H.Y., 82503802 to B.C.), the Fundamental Research Funds for the Central Universities (2042022dx0003 to J.Z., 2042021kf0075 to H.Y.), the Postdoctoral Fellowship Program of CPSF (GZC20241276 to W.X., GZC20250976 to C.H.), the China Postdoctoral Science Foundation (2022M722462 to H.L., 2024M762491 to C.H., 2025M772321 to W.X.), the Postdoctoral Project of Hubei Province (2004HBBHJD057 to C.H.), Natural Science Foundation of Wuhan (2024040701010031 to J.Z.), the Open Projects of Hubei Key Laboratory of Tumor Biological Behavior (220172107 to J.Z.), the Natural Science Foundation of Hubei Province of China (2022CFA008 to J.Z.), the Open Research Fund of the National Center for Protein Sciences at Peking University in Beijing (KF-202504 to J.Z.), the Natural Science Foundation Youth Category A of Hubei Province (JCZRJQ202500008 to H.Z.), the Natural Science Foundation of Chongqing, China (CSTB2024NSCQ-MSX0066 to H.Z.), the translational Medicine and Interdisciplinary Research Joint Fund of Zhongnan Hospital of Wuhan University (ZNJC202312 to J.Z.), and USA National Institutes of Health P50 CA101942 (G.J.F.).

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Author notes

  1. These authors contributed equally: Yishuang Sun, Zijian Zhang, Haiou Li, Xia Bu, Li Chen.

Authors and Affiliations

  1. Department of Hepatobiliary and Pancreatic Surgery, State Key Laboratory of Metabolism and Regulation in Complex Organisms, Medical Research Institute, Frontier Science Center for Immunology and Metabolism, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, Hubei, China

    Yishuang Sun, Xiyong Wang, Lijun Kong, Wenjing Song, Xiangling Xiao, Jie Shi, Bolin Xiang, Chuan He, Yingmeng Yao, Wenjun Xiong, Haisheng Yu, Hudan Liu, Guoliang Qing, Zhiyong Yang & Jinfang Zhang

  2. Department of Pathology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China

    Yishuang Sun, Lifang Fan & Sufang Tian

  3. Taikang Center for Life and Medical Sciences, Wuhan University, Wuhan, Hubei, China

    Yishuang Sun, Xiyong Wang, Lijun Kong, Xiangling Xiao, Jie Shi, Bolin Xiang, Chuan He, Yingmeng Yao, Wenjun Xiong, Haisheng Yu, Hudan Liu, Guoliang Qing & Jinfang Zhang

  4. Key Laboratory of Tropical Biological Resources of Ministry of Education, School of Pharmaceutical Sciences, Hainan University, Haikou, Hainan, China

    Zijian Zhang

  5. National “111” Center for Cellular Regulation and Molecular Pharmaceutics, Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei University of Technology, Wuhan, Hubei, China

    Zijian Zhang & Haichuan Zhu

  6. Department of Dermatology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China

    Haiou Li

  7. Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA

    Xia Bu & Gordon J. Freeman

  8. Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

    Li Chen & Wenyi Wei

  9. Department of Colorectal and Anal Surgery, Low Rectal Cancer Diagnosis and Treatment Center, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China

    Baoxiang Chen, Congqing Jiang & Qun Qian

  10. Department of Radiation and Medical Oncology, Hubei Key Laboratory of Tumor Biological Behaviors, Hubei Provincial Clinical Research Center for Cancer, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China

    Panpan Dai & Jinfang Zhang

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  1. Yishuang Sun
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Contributions

Y.S. performed most of the experiments with help from Z.Z., Haiou L., X.B., L.C., X.W., L.K., P.D., W.S., X.X., J.S., B.X., C.H., Y.Y., W.X., and H.Y.; J.Z., Y.S., G.J.F., H.Z., and W.W. designed experiments; L.F., B.C., C.J., and Q.Q. collected human colorectal cancer tissues; X.B. and G.J.F. provided essential reagents; J.Z., H.Z., G.J.F., and W.W. guided and supervised the project. J.Z. and Y.S. wrote the manuscript. X.B., Z.Y., G.Q., S.T., Hudan L., W.W., and G.J.F. provided discussion and edited the manuscript. All authors commented on the manuscript.

Corresponding authors

Correspondence to Wenyi Wei, Gordon J. Freeman, Haichuan Zhu or Jinfang Zhang.

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Competing interests

G.J.F. has patents/royalties on the PD-L1/PD-1 pathway from Bristol-Myers-Squibb, Roche, Eli Lilly, and Novartis. G.J.F. has served on advisory boards for iTeos, NextPoint, IgM, GV20, IOME, Bioentre, Santa Ana Bio, Simcere of America, and Geode. G.J.F. has equity in Nextpoint, Triursus, Xios, iTeos, IgM, Invaria, GV20, Bioentre, and Geode. W.W. is a co-founder and consultant for the ReKindle Therapeutics. The other authors declare no competing interests.

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Sun, Y., Zhang, Z., Li, H. et al. Destruction of VISTA by TRIM25 ablation in T cells potentiates cancer immunotherapy. Cell Res 35, 1003–1020 (2025). https://doi.org/10.1038/s41422-025-01186-5

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