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Molecular Diagnostics

Peripheral blood neutrophils contribute to Claudin18.2-specific CAR-T cell treatment resistance in advanced gastric cancer

British Journal of Cancer volume 132, pages 1167–1176 (2025)Cite this article

Subjects

Abstract

Background

Claudin18.2 (CLDN18.2)-specific chimeric antigen receptor (CAR)-T cell treatment holds promise for advanced gastric cancer (GC) but has variable efficacy. This study investigates the prognostic value of the neutrophil-to-lymphocyte ratio (NLR) in CAR-T cell treatment and elucidates the molecular mechanisms of treatment resistance.

Methods

GC patients treated with CLDN18.2-specific CAR-T cell treatment were analyzed. Outcomes included objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS), and overall survival (OS). Survival analyses utilized Kaplan-Meier methods, log-rank tests, and Cox regression. Single-cell RNA sequencing was performed on peripheral blood samples to investigate the mechanisms of pro-tumor circulating neutrophils.

Results

Elevated NLR was significantly associated with lower ORR (34.2% vs. 55.9%, P < 0.001), shorter median PFS (3.6 vs. 8.0 months, P < 0.001), and OS (5.6 vs. 13.8 months, P < 0.001). Single-cell sequencing identified a circulating neutrophil subcluster (NE-3) linked to disease progression. NE-3 expressed pro-tumoral factors (MMP-9), and was enriched in the IL-17 signaling pathway. The cellular interactions between neutrophils and T cells were more prominent in progression disease (PD) group than in partial response (PR) group.

Conclusions

This study highlights NLR as a significant prognostic factor in advanced GC patients receiving CLDN18.2-specific CAR-T cell treatment and provides insights into neutrophil-mediated treatment resistance. Further validation and exploration of strategies to mitigate neutrophil-induced immunosuppression are needed.

Trial Registration

NCT03874897.

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Fig. 1: Association between neutrophil to lymphocyte ratio (NLR) and the clinical outcomes of advanced gastric cancer patients receiving CLDN18.2-specific CAR-T cell therapy.
Fig. 2: Forest plot of univariate and multivariate analysis of clinicopathological characteristics associated with survival.
Fig. 3: Cellular profiling in peripheral blood samples.
Fig. 4: Cellular subcluster analysis of neutrophils in peripheral blood samples.
Fig. 5: Cellular interaction analysis among different cell types in peripheral blood.

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Data availability

The single-cell RNA sequencing data have been deposited at China National Center for Bioinformation with the number of HRA006971. Requests for individual participant-level data from this study should be submitted via email to the corresponding author with detailed proposals.

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Acknowledgements

We thank all patients and their families in this trial.

Funding

This study was funded by the National Key Research and Development Program of China (No. 2022YFC2505006, No. 2023YFC3403700, and No. 2022YFA0912400), National Natural Science Foundation of China (No. U22A20327), Beijing Natural Science Foundation (L232080), Beijing Hospitals Authority Youth Program (QMS20201101), Science Foundation of Peking University Cancer Hospital (JC202406), Clinical Medicine Plus X - Young Scholars Project of Peking University, Peking University Clinical Scientist Training Program, Fundamental Research Funds for the Central Universities, and CARsgen Therapeutics Co., Ltd.

Author information

Author notes

  1. These authors contributed equally: Jiarui Li, Min Tao, Lian Liu.

Authors and Affiliations

  1. Beijing Key Laboratory of Cell & Gene Therapy for Solid Tumor, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Gastrointestinal Oncology, Peking University Cancer Hospital & Institute, Beijing, China

    Jiarui Li, Min Tao, Lian Liu & Mingyang Ma

  2. Beijing Key Laboratory of Cell & Gene Therapy for Solid Tumor, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Early Drug Development Centre, Peking University Cancer Hospital & Institute, Beijing, China

    Chang Liu, Dan Liu, Panpan Zhang, Miao Zhang & Ran Xue

  3. Beijing Key Laboratory of Cell & Gene Therapy for Solid Tumor, State Key Laboratory of Holistic Integrative Management of Gastrointestinal Cancers, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Gastrointestinal Oncology, Peking University Cancer Hospital & Institute, Beijing, China

    Jifang Gong, Cheng Zhang, Xiaotian Zhang & Lin Shen

  4. Beijing Key Laboratory of Cell & Gene Therapy for Solid Tumor, State Key Laboratory of Holistic Integrative Management of Gastrointestinal Cancers, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Early Drug Development Centre, Peking University Cancer Hospital & Institute, Beijing, China

    Changsong Qi

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Contributions

JL, LL, MT, CL, MM, DL, PZ, MZ, RX, JG, CZ, and CQ collected the data. JL and MM performed the statistical analyses. CQ, XZ, and LS conceived and designed the trial. JL drafted the manuscript. CQ, XZ, and LS reviewed and revised the manuscript. All authors read and approved the final version of the manuscript.

Corresponding authors

Correspondence to Xiaotian Zhang, Lin Shen or Changsong Qi.

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The authors declare no competing interests.

Ethics approval and consent to participate

The study was approved by the Ethics Committee of Peking University Cancer Hospital (2018YJZ75). The study was performed in accordance with the Declaration of Helsinki and Good Clinical Practice guidelines. All patients provided written informed consent before participation in this trial.

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Li, J., Tao, M., Liu, L. et al. Peripheral blood neutrophils contribute to Claudin18.2-specific CAR-T cell treatment resistance in advanced gastric cancer. Br J Cancer 132, 1167–1176 (2025). https://doi.org/10.1038/s41416-025-03015-3

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