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Unsynchronized butyrophilin molecules dictate cancer cell evasion of Vγ9Vδ2 T-cell killing

Cellular & Molecular Immunology volume 21, pages 362–373 (2024)Cite this article

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Abstract

Vγ9Vδ2 T cells are specialized effector cells that have gained prominence as immunotherapy agents due to their ability to target and kill cells with altered pyrophosphate metabolites. In our effort to understand how cancer cells evade the cell-killing activity of Vγ9Vδ2 T cells, we performed a comprehensive genome-scale CRISPR screening of cancer cells. We found that four molecules belonging to the butyrophilin (BTN) family, specifically BTN2A1, BTN3A1, BTN3A2, and BTN3A3, are critically important and play unique, nonoverlapping roles in facilitating the destruction of cancer cells by primary Vγ9Vδ2 T cells. The coordinated function of these BTN molecules was driven by synchronized gene expression, which was regulated by IFN-γ signaling and the RFX complex. Additionally, an enzyme called QPCTL was shown to play a key role in modifying the N-terminal glutamine of these BTN proteins and was found to be a crucial factor in Vγ9Vδ2 T cell killing of cancer cells. Through our research, we offer a detailed overview of the functional genomic mechanisms that underlie how cancer cells escape Vγ9Vδ2 T cells. Moreover, our findings shed light on the importance of the harmonized expression and function of gene family members in modulating T-cell activity.

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Acknowledgements

We thank members of the Wei laboratory for their comments, technical support, and experimental assistance. We thank Dr. Ying Liu for providing the A375-Cas9 cells.

Funding

WW received funding from the National Science Foundation of China (31930016) and the Peking-Tsinghua Center for Life Sciences. ZW received funding from the State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases (2024KF00001) and the National Science Foundation of China (82350119). CCW received funding from the Talent Introduction Funds from the Chinese Academy of Medical Science (2022-RC310-10), the National Science Foundation of China (32150005), and the Research Funds from Health@InnoHK Program, launched by the Innovation Technology Commission of the Hong Kong Special Administrative Region.

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Author notes

  1. These authors contributed equally: Zeguang Wu, Qiezhong Lamao, Meichao Gu

Authors and Affiliations

  1. Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, 100871, Beijing, China

    Zeguang Wu, Qiezhong Lamao, Meichao Gu, Xuanxuan Jin, Ying Liu, Feng Tian, Ying Yu & Wensheng Wei

  2. EdiGene Inc., Life Science Park, Changping District, 102206, Beijing, China

    Pengfei Yuan

  3. Center for Precision Medicine Multi-Omics Research, Peking University Health Science Center, Peking University, 100191, Beijing, China

    Shuaixin Gao

  4. Department of Microbiology and Immunology at the Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Parkville, VIC, 3010, Australia

    Thomas S. Fulford & Adam P. Uldrich

  5. Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia

    Adam P. Uldrich

  6. State Key Laboratory for Complex, Severe and Rare Diseases, Clinical Research Institute, Peking Union Medical College Hospital, Chinese Academy of Medical Science & Peking Union Medical College, Peking-Tsinghua Center for Life Sciences, 100871, Beijing, China

    Catherine CL Wong

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Contributions

WW, ZW, QL, and MG designed and performed the research, analyzed the data and wrote the manuscript. XJ and YL performed the bioinformatics analyses. YY and FT performed the research and analyzed the data. PY provided PBMCs from healthy donors and the sgRNA library. SG and CCW performed the mass spectrometry analysis. TSF and APU provided the TCR tetramers and detection methods. WW and CCW supervised the project. All the authors discussed the results and contributed to the final manuscript.

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Correspondence to Catherine CL Wong or Wensheng Wei.

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WW is a founder and scientific adviser for EdiGene and Therorna, Inc. The other authors declare no competing interests.

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Wu, Z., Lamao, Q., Gu, M. et al. Unsynchronized butyrophilin molecules dictate cancer cell evasion of Vγ9Vδ2 T-cell killing. Cell Mol Immunol 21, 362–373 (2024). https://doi.org/10.1038/s41423-024-01135-z

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