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A VEGFR2–MICA bispecific antibody activates tumor-infiltrating lymphocytes and exhibits potent anti-tumor efficacy in mice

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Abstract

MHC class I-related chain A (MICA) is one of the major ligands for natural killer group 2 member D (NKG2D), which is an activating NK receptor. MICA is expressed on the surface of human epithelial tumor cells, and its shedding from tumor cells leads to immunosuppression. To activate immune response in the tumor microenvironment, we designed an anti-VEGFR2–MICA bispecific antibody (JZC01), consisting of MICA and an anti-VEGFR2 single chain antibody fragment (JZC00) and explored its potential anti-tumor activity. JZC01 targeted vascular endothelial growth factor receptor 2 (VEGFR2) and inhibited tumorigenesis by blocking the VEGFR2 signaling pathway. Additionally, JZC01 promoted NK and CD8+ T cells to release IFN-γ and engaged activated lymphocytes to lysis of VEGFR2-expressing tumor cells. The in vivo anti-tumor activity of JZC01 was investigated by establishing a Lewis lung cancer cell-transplanted mouse model. It effectively reduced the tumor vascular density and increased the infiltration and activation of NK and CD8+ T cells in the tumor microenvironment. Thus, JZC01 functions in anti-tumor angiogenesis and anti-tumor immune activation, and showed improved anti-tumor efficacy combined with docetaxel, which provides a new insight into anti-tumor therapy.

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Abbreviations

JZC00:

Anti-VEGFR2 single chain antibody

JZC01:

Anti-VEGFR2–MICA bispecific antibody

MICA:

MHC class I-related chain A

NKG2D:

Natural killer group 2 member D

NSCLC:

Non-small cell lung cancer

rMICA:

Recombined human MICA

rNKG2D:

Recombined human NKG2D

TIL:

Tumor-infiltrating lymphocytes

VEGF:

Vascular endothelial growth factor

VEGFR2:

Vascular endothelial growth factor receptor 2

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Funding

This work was supported in part by the National Natural Science Foundation of China (NSFC 81973223), Natural Science Foundation of Jiangsu Province (BK20161459), Jiangsu Province Qinglan Project (2014); “Double First-Class” University project (CPU2018PZQ12 and CPU2018GY14).

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Authors and Affiliations

  1. Antibody Engineering Laboratory, State Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical University, 154#, Tong Jia Xiang 24, Nanjing, 210009, People’s Republic of China

    Yao Xu, Xinrong Zhang, Mingzhu Pan, Min Wang & Juan Zhang

  2. Sanhome-CPU Joint Laboratory, China Pharmaceutical University, Nanjing, 210009, People’s Republic of China

    Yao Xu & Yong Wang

Authors
  1. Yao Xu
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  2. Xinrong Zhang
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  3. Yong Wang
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  4. Mingzhu Pan
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  5. Min Wang
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  6. Juan Zhang
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Contributions

YX and XZ designed and performed the experiments. YW guided the affinity measurement and cell-based binding assay. MP helped with the NK and CD8+ T cells isolation and IFN-γ detection. JZ, MW, and YW designed the study. YX and JZ wrote the paper.

Corresponding author

Correspondence to Juan Zhang.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval and ethical standards

This article does not contain any studies with human participants performed by any of the authors. This study was approved by the Ethics Review Committee of China Pharmaceutical University (9 March 2016). All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. Six-week-old c57BL/6 healthy female mice were purchased from Comparative Medicine Centre of Yangzhou University (Yangzhou, China). All animals were housed in a pathogen-free environment and were treated following the standards of Pharmaceutical Animal Center of China Pharmaceutical University (Approval number XYXK-2016-0011).

Cell line authentication

Murine cancer cell lines, Lewis lung carcinoma (LLC) (CRL-1642, ATCC), and mouse forestomach gastric carcinoma (MFC) (100207, BNCC) were obtained from Shanghai cell bank, Chinese Academy of Sciences. No cell line authentication was necessary since the phenotypes of these cells were checked by Shanghai cell bank before we obtained them and they did not change over time.

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Xu, Y., Zhang, X., Wang, Y. et al. A VEGFR2–MICA bispecific antibody activates tumor-infiltrating lymphocytes and exhibits potent anti-tumor efficacy in mice. Cancer Immunol Immunother 68, 1429–1441 (2019). https://doi.org/10.1007/s00262-019-02379-9

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