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Dual CAR-T cells to treat cancers co-expressing NKG2D and PD1 ligands in xenograft models of peritoneal metastasis

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Abstract

While the expression of either NKG2D ligands or PD-1 ligands has been reported in various types of cancers, the co-expression of the two sets of ligands in the same tumour tissues is still un-investigated. After examining 68 primary ovarian cancer samples, we observed around 80% of the co-expression in low grade serous and endometrioid ovarian cancer samples. We then constructed a dual CAR system that splits the conventional single-input of a 2nd generation CAR into two independent chimeric receptors, one composed of the NKG2D extracellular domain linked with DAP12 for T cell activation and another using the PD-1 extracellular domain linked with 4-1BB for costimulatory signal 2 input. Given the limitation of the low-affinity PD-1 receptor in recognizing cancer cells with low levels of PD-1 ligands, we also used a high-affinity scFv specific to PD-L1 in our combinatorial approach to expand the range of target cancer cells with different expression levels of PD-L1. The two types of dual CAR-T cells were generated through electroporation of non-viral piggyBac transposon plasmids and were effective in eliminating the target cancer cells. Especially, the dual CAR-T cells with anti-PD-L1 scFv were capable of eradicating established tumors in mouse models of peritoneal metastasis of colorectal cancer and ovarian cancer. Since both NKG2D ligands and PD-1 ligands have been marked as favourable cancer therapeutic targets, the new dual CAR-T cells developed in this study hold attractive application potential in treating metastatic peritoneal carcinoma.

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Data availability

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

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Funding

This work was supported by the Singapore Ministry of Health’s National Medical Research Council (NMRC/OFLCG/003/2018; MOH-000465-01); Agency for Science, Technology and Research, Singapore (IAF-PP:H19/01/a0/022); and by Cheetah Cell Therapeutics.

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Author notes

  1. Guangyi Jiang, Yu Yang Ng and Johan C. K. Tay have contributed equally to the work.

Authors and Affiliations

  1. Department of Gynecological Oncology, Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou, 310022, People’s Republic of China

    Guangyi Jiang, Shu Wang & Jianqing Zhu

  2. Department of Biological Sciences, National University of Singapore, Singapore, 117543, Singapore

    Yu Yang Ng, Johan C. K. Tay, Zhicheng Du & Shu Wang

  3. Cheetah Cell Therapeutics, #501, No 2 Avenue, Hangzhou, 310018, People’s Republic of China

    Lin Xiao

Authors
  1. Guangyi Jiang
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  2. Yu Yang Ng
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  4. Zhicheng Du
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Contributions

SW and JZ contributed to the study conception and design. Material preparation, data collection and analysis were performed by GJ, YYN, JCKT, LX and ZD. The first draft of the manuscript was written by GJ, YYN and JCKT. SW and JZ commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Jianqing Zhu.

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Conflict of interest

The authors Shu Wang, Yu Yang Ng, Johan CK Tay have filed patent applications related to CAR technology and could potentially receive licensing royalties in future. Lin Xiao is employee of Cheetah Cell Therapeutics.

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Jiang, G., Ng, Y.Y., Tay, J.C.K. et al. Dual CAR-T cells to treat cancers co-expressing NKG2D and PD1 ligands in xenograft models of peritoneal metastasis. Cancer Immunol Immunother 72, 223–234 (2023). https://doi.org/10.1007/s00262-022-03247-9

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  • DOI: https://doi.org/10.1007/s00262-022-03247-9

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