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EGFRvIII-CAR-T Cells with PD-1 Knockout Have Improved Anti-Glioma Activity

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Pathology & Oncology Research

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Abstract

Glioblastoma multiforme (GBM) is the most malignant form of the brain tumors. EGFR variant III (EGFRvIII) is expressed in about 30% of GBM specimens, but not expressed in normal brain tissues. Therefore, EGFRvIII protein offers an ideal CAR-T therapeutic target for EGFRvIII-positive GBM patients. PD-L1 is expressed in a variety of cancer cells, including GBM. Tumor-associated PD-L1 can bind to PD-1 on T cells and promote apoptosis of T cells, thus suppressing the anti-cancer immune response. In our current studies, PD-1WT EGFRvIII-CAR-T cells and PD-1KD EGFRvIII-CAR-T cells were generated. Cytokine production and lytic activity of these two CAR-T cells against to PD-L1WT EGFRvIII+ U373 cells or PD-L1KO EGFRvIII+ U373 cells were evaluated. The results showed that PD-1KD EGFRvIII-CAR-T cells and PD-1WT EGFRvIII-CAR-T cells showed same levels of interferon-γ (IFN-γ) and interleukin-2 (IL-2) production as well as cytolytic activity against PD-L1KO EGFRvIII+ U373 cells; however, PD-1KD EGFRvIII-CAR-T cells exhibited higher levels of IFN-γ and IL-2 production as well as cytolytic activity against PD-L1+ EGFRvIII+ U373 cells than that of PD-1WT EGFRvIII-CAR-T cells. PD-1KD EGFRvIII-CAR-T cells also exhibited higher anti-glioma activity and longer survival in mice in vivo than that of PD-1WT EGFRvIII-CAR-T cells. Taken together, our findings indicate that PD-1 knockout enhances lytic activity of EGFRvIII-CAR-T cells against PD-L1+ EGFRvIII+ GBM cells. These might provide a new insight into strategy of GBM CAR-T cell therapy.

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Acknowledgements

We would like to thank Aolin He for his technical assistance in FCS analysis.

Funding

This work was supported by National Natural Science Foundation of China (81772691 and 81370062), China postdoctoral science foundation grant (2017 M620196 and 2018 T110467), and Key Young Medical Talents Project in Jiangsu Province (QNRC2016526). The funders had no role in the study design, data collection and analysis, decision to publish, or in the preparation of the manuscript.

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Authors and Affiliations

  1. Department of Neurosurgery, Nan**g Medical University, Nan**g, 210029, People’s Republic of China

    Haifeng Zhu

  2. Department of Neurosurgery, Funing People’s Hospital, Funing, 224400, People’s Republic of China

    Haifeng Zhu & Zhouming Shen

  3. Department of Neurosurgery, The First Affiliated Hospital of Nan**g Medical University, Nan**g, 210029, People’s Republic of China

    Yong** You

  4. epartment of Neurosurgery, Affiliated Kunshan Hospital of Jiangsu University, Suzhou, 215300, People’s Republic of China

    Lei Shi

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  1. Haifeng Zhu
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Correspondence to Yong** You or Lei Shi.

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The authors declare that there is no conflict of interest.

Ethical Approval

All procedures performed in this study were in accordance with the ethical standards of the Affiliated Kunshan Hospital of Jiangsu University Ethics Committee for Scientific Research (No. 201901276) and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Zhu, H., You, Y., Shen, Z. et al. EGFRvIII-CAR-T Cells with PD-1 Knockout Have Improved Anti-Glioma Activity. Pathol. Oncol. Res. 26, 2135–2141 (2020). https://doi.org/10.1007/s12253-019-00759-1

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  • DOI: https://doi.org/10.1007/s12253-019-00759-1

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