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Noninvasive longitudinal PET/CT imaging of CAR T cells using PSMA reporter gene

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European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

Chimeric antigen receptor (CAR) T cell therapy has achieved great success in treating hematologic malignancies. However, it is yet to prove effective in the treatment of solid tumors. Thus, it is necessary to develop appropriate methodology for the long-term, accurate, and quantitative evaluation of the distribution and activities of CAR T cells in solid tumors. In the present study, we engineered TfR ΔPSMA CAR (CAR-ΔPSMA) T cells, which targeted the transferrin receptor (TfR) expressed by tumor cells and could be tracked in vivo via a reporter gene encoding the truncated prostate specific membrane antigen (ΔPSMA). We then quantitatively monitored these CAR T cells in vitro and in vivo using [68Ga]Ga-PSMA-617 positron emission tomography (PET)/computed tomography (CT).

Methods

The CAR-ΔPSMA T cells were genetically engineered by transducing T cells with a lentiviral vector encoding TfR41BBζ-T2A-ΔPSMA. Firstly, the target expression, activation, and cytotoxicity of CAR-ΔPSMA T cells were validated in vitro. Secondly, the minimum thresholds of CAR-ΔPSMA T cells detection for [68Ga]Ga-PSMA-617 PET/CT were also determined in vitro and in vivo respectively. Lastly, the feasibility of monitoring the biodistribution and infiltration of CAR-ΔPSMA T cells after systematic administration was evaluated in the breast cancer subcutaneous xenograft model.

Results

The CAR-ΔPSMA T cells retained activation and tumor killing capacity after transduction of the ΔPSMA-encoding reporter gene. Next, the CAR-ΔPSMA T cells could be reliably tracked by [68Ga]Ga-PSMA-617 PET/CT, the detection sensitivity of which was 250 cells/mm3 in vitro and 100 cells/mm3 in vivo. Next, the sequential imaging assays revealed that [68Ga]Ga-PSMA-617 PET/CT could be used to specifically visualize ΔPSMA+ CAR T cells at the tumor site. The increase in the [68Ga]Ga-PSMA-617 signal intensity over time allowed us to effectively detect CAR T cells in vivo.

Conclusion

Our findings preliminarily confirmed that [68Ga]Ga-PSMA-617 PET/CT could reliably detect CAR-ΔPSMA T cells in vitro and in vivo in solid tumors, laying the foundation for the monitoring CAR T cell therapy in the future.

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Data availability

Data and materials are available from the corresponding authors upon reasonable request.

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Funding

This work was financially supported by National Natural Science Foundation of China (grants 82030052), Hubei Province Science and Technology Innovation Team ([2022] No. 72) and Key Project of Hubei Province Natural Science Foundation (No. 2021CFA008).

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Author notes

  1. Xiangming Song and Yirui Zhang contributed equally to this work.

Authors and Affiliations

  1. Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1277 Jiefang Ave, Wuhan, China

    Xiangming Song, Xiaoying Lv, Yu Long, Yongkang Gai, Dawei Jiang & Xiaoli Lan

  2. Hubei Province Key Laboratory of Molecular Imaging, Wuhan, 430022, Hubei Province, China

    Xiangming Song, Xiaoying Lv, Yu Long, Yongkang Gai, Dawei Jiang & Xiaoli Lan

  3. Key Laboratory of Biological Targeted Therapy, The Ministry of Education, Wuhan, China

    Xiangming Song, Xiaoying Lv, Yu Long, Yongkang Gai, Dawei Jiang & Xiaoli Lan

  4. Department of Immunology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

    Yirui Zhang, Zhuoshuo Xu & Ping Lei

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  1. Xiangming Song
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Contributions

X. L., L. P and D. J.: study design; X. S. and Y. Z.: manuscript writing; X. S. and Y. G.: tracer synthesis and identification; Y. Z. and Z. X.: Preparation of CAR T cells; X. S., X. L. and Y. L.: image acquisition and interpretation; X. L., L. P and D. J.: manuscript revision.

Corresponding author

Correspondence to Xiaoli Lan.

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All experimental schemes were performed under the guidance and approved by Union Hospital, Tongji Medical College, Huazhong University of Science and Technology and Institutional Animal Care and Use Committee of Tongji Medical College of Huazhong University of Science and Technology.

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Song, X., Zhang, Y., Lv, X. et al. Noninvasive longitudinal PET/CT imaging of CAR T cells using PSMA reporter gene. Eur J Nucl Med Mol Imaging 51, 965–977 (2024). https://doi.org/10.1007/s00259-023-06508-6

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