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Evaluation of a xenogeneic vascular endothelial growth factor-2 vaccine in two preclinical metastatic tumor models in mice

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Abstract

In this study, a xenogeneic DNA vaccine encoding for human vascular endothelial growth factor receptor-2 (hVEGFR-2) was evaluated in two murine tumor models, the B16-F10 melanoma and the EO771 breast carcinoma model. The vaccine was administered by intradermal injection followed by electroporation. The immunogenicity and the biological efficacy of the vaccine were tested in (1) a prophylactic setting, (2) a therapeutic setting, and (3) a therapeutic setting combined with surgical removal of the primary tumor. The tumor growth, survival, and development of an immune response were followed. The cellular immune response was measured by a bioluminescence-based cytotoxicity assay with vascular endothelial growth factor-2 (VEGFR-2)-expressing target cells. Humoral immune responses were quantified by enzyme-linked immunosorbent assay (ELISA). Ex vivo bioluminescence imaging and immunohistological observation of organs were used to detect (micro)metastases. A cellular and humoral immune response was present in prophylactically and therapeutically vaccinated mice, in both tumor models. Nevertheless, survival in prophylactically vaccinated mice was only moderately increased, and no beneficial effect on survival in therapeutically vaccinated mice could be demonstrated. An influx of CD3+ cells and a slight decrease in VEGFR-2 were noticed in the tumors of vaccinated mice. Unexpectedly, the vaccine caused an increased quantity of early micrometastases in the liver. Lung metastases were not increased by the vaccine. These early liver micrometastases did however not grow into macroscopic metastases in either control or vaccinated mice when allowed to develop further after surgical removal of the primary tumor.

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Abbreviations

CTC:

Circulating tumor cells

eGFP:

Enhanced green fluorescent protein

hVEGFR-2:

Human vascular endothelial growth factor-2

IVIS:

In vivo imaging system

PBS:

Phosphate buffered saline

pDNA:

Plasmid DNA

VEGFR-2:

Vascular endothelial growth factor-2

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Acknowledgements

This work was supported by a grant of Ghent University and by grants from the Fonds Wetenschappelijk onderzoek (G.0235.11N and G.0621.10N). The technical assistance of Sarah Loomans was greatly appreciated (Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University).

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Author notes

  1. Sofie Denies

    Present address: iTeos Therapeutics, Rue Clément Ader 16, 6041, Gosselies, Belgium

  2. Laetitia Cicchelero

    Present address: Bimetra, Clinical Research Centre Ghent, De Pintelaan 185, 9000, Ghent, Belgium

  3. Sofie Denies and Bregje Leyman contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Gene Therapy, Department of Nutrition, Genetics and Ethology, Faculty of Veterinary Medicine, Ghent University, Heidestraat 19, 9820, Merelbeke, Belgium

    Sofie Denies, Bregje Leyman, Hanne Huysmans, Francis Combes, Séan Mc Cafferty, Laetitia Cicchelero, Joyca De Temmerman & Niek N. Sanders

  2. Cancer Research Institute Ghent (CRIG), Ghent University, 9000, Ghent, Belgium

    Francis Combes, Séan Mc Cafferty & Niek N. Sanders

  3. Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820, Merelbeke, Belgium

    Marjan Steppe

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Contributions

SD and NNS conceived and designed the experiments. SD performed the experiments and together with NNS drafted the initial submission. BL managed the revision of the paper and together with NNS drafted the revised manuscript. HH and JDT assisted with the immunizations and the follow-up of the mice during the revision. SMC produced the pDNA vaccine and inoculated the mice during the revision. LC was involved in the immunizations of the initial submission. FC and MS assisted with the histology and immunohistochemistry.

Corresponding author

Correspondence to Niek N. Sanders.

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The authors declare that they have no conflict of interest.

Ethical approval

All experiments involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. The experiments were approved by the ethical committee of the Faculty of Veterinary Medicine (Ghent University, Belgium; EC-DI 2014-45).

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No patients were involved in this study.

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Denies, S., Leyman, B., Huysmans, H. et al. Evaluation of a xenogeneic vascular endothelial growth factor-2 vaccine in two preclinical metastatic tumor models in mice. Cancer Immunol Immunother 66, 1545–1555 (2017). https://doi.org/10.1007/s00262-017-2046-3

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