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Immuno-PET imaging of 68Ga-labeled nanobody Nb109 for dynamic monitoring the PD-L1 expression in cancers

Abstract

The checkpoint blockade immunotherapy has become a potent treatment strategy for cancers, and programmed death ligand-1 (PD-L1) is a prominent checkpoint ligand that is highly expressed in some cancers. The identification of immune checkpoint marker PD-L1 is critical for improving the success of immunotherapy. Accordingly, the binding specificity and dynamic monitoring property of a non-blocking nanobody tracer 68Ga-NOTA-Nb109 to PD-L1 were assessed in this study. The endogenous expression level of PD-L1 in several cancer cells was measured by flow cytometry, Western blot, and cellular uptake assay. Sensitivity and specificity of 68Ga-NOTA-Nb109 in monitoring the expression of PD-L1 in vivo were evaluated by PET imaging of different tumor-bearing models (U87, high PD-L1 expression; HCT 116, medium PD-L1 expression; and NCI-H1299, low PD-L1 expression). In vivo PET imaging results agreed well with those detected in vitro. In addition, PET imaging of PD-L1 expression in U87 and NCI-H1299 xenografts using 18F-FDG was also performed for comparison. The maximum tumor-to-muscle uptake ratio of 68Ga-NOTA-Nb109 was more than twofold that of 18F-FDG in U87 xenograft. The change of PD-L1 expression in NCI-H1299 cells and xenografts induced by cisplatin (CDDP) was sensitively monitored by 68Ga-NOTA-Nb109. This study demonstrated the feasibility of tracer 68Ga-NOTA-Nb109 for specifically targeting endogenous PD-L1 and dynamic monitoring the change of PD-L1 expression, and could guide the immunotherapy and immunochemotherapy for refractory cancers.

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Abbreviations

CDDP:

Cisplatin

IHC:

Immunohistochemistry

mAbs:

Monoclonal antibodies

NSCLC:

Non-small cell lung cancer

PD-1:

Programmed death protein 1

PD-L1:

Programmed death ligand-1

ROI:

Region of interest

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Acknowledgements

The nanobody of NOTA-Nb109 was kindly provided by Suzhou Smart Nuclide Biopharmaceutical Co., Ltd, China.

Funding

This work was funded by the National Natural Science Foundation of China (81972906, 81971645, and 22076069), Natural Science Foundation of Jiangsu Province (BK20181128 and BK20201135), Key Youth Medical Talent Project of Jiangsu Province (QNRC2016626 and QNRC2016629), Jiangsu Commission of Health (ZDA2020007 and M2020028), Precision Medical Project of Wuxi Commission of Health and Family Planning (J201806), Major Scientific Research Project of Wuxi Commission of Health (Z201913), and Innovation Capacity Development Plan of Jiangsu Province (BM2018023).

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Authors

Contributions

QL and LJ performed the experiments, acquired and analyzed the data. QL wrote the paper, and LJ contributed to paper writing. KL and GL contributed to the PET imaging, and analyzed data for in vivo experiments. HL performed the probe labeling and quality control. JL and LQ conceived, designed, and supported the study, interpreted the results and revised the manuscript.

Corresponding authors

Correspondence to Jianguo Lin or Ling Qiu.

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The authors declare that they have no conflict of interest.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All animal procedures were performed according to the protocols approved by the ethical committee of Jiangsu Institute of Nuclear Medicine.

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Liu, Q., Jiang, L., Li, K. et al. Immuno-PET imaging of 68Ga-labeled nanobody Nb109 for dynamic monitoring the PD-L1 expression in cancers. Cancer Immunol Immunother 70, 1721–1733 (2021). https://doi.org/10.1007/s00262-020-02818-y

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Keywords

  • PD-L1
  • PET imaging
  • Nanobody tracer
  • 68Ga
  • 18F-FDG