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Intratumoral DNA-based delivery of checkpoint-inhibiting antibodies and interleukin 12 triggers T cell infiltration and anti-tumor response

Cancer Gene Therapy volume 29, pages 984–992 (2022)Cite this article

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Abstract

To improve the anti-tumor efficacy of immune checkpoint inhibitors, numerous combination therapies are under clinical evaluation, including with IL-12 gene therapy. The current study evaluated the simultaneous delivery of the cytokine and checkpoint-inhibiting antibodies by intratumoral DNA electroporation in mice. In the MC38 tumor model, combined administration of plasmids encoding IL-12 and an anti-PD-1 antibody induced significant anti-tumor responses, yet similar to the monotherapies. When treatment was expanded with a DNA-based anti-CTLA-4 antibody, this triple combination significantly delayed tumor growth compared to IL-12 alone and the combination of anti-PD-1 and anti-CTLA-4 antibodies. Despite low drug plasma concentrations, the triple combination enabled significant abscopal effects in contralateral tumors, which was not the case for the other treatments. The DNA-based immunotherapies increased T cell infiltration in electroporated tumors, especially of CD8+ T cells, and upregulated the expression of CD8+ effector markers. No general immune activation was detected in spleens following either intratumoral treatment. In B16F10 tumors, evaluation of the triple combination was hampered by a high sensitivity to control plasmids. In conclusion, intratumoral gene electrotransfer allowed effective combined delivery of multiple immunotherapeutics. This approach induced responses in treated and contralateral tumors, while limiting systemic drug exposure and potentially detrimental systemic immunological effects.

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Fig. 1: Combined intratumoral delivery of DNA-based IL-12 and a DNA-based anti-PD-1 antibody in MC38 tumors.
Fig. 2: Combined intratumoral delivery of DNA-based IL-12 and DNA-based anti-PD-1 and anti-CTLA-4 antibodies in MC38 tumors.
Fig. 3: Effect of intratumoral delivery of DNA-based IL-12, anti-PD-1 and anti-CTLA-4 antibodies on MC38-infiltrating T lymphocytes.
Fig. 4: Combined intratumoral delivery of DNA-based IL-12 and DNA-based anti-PD-1 and anti-CTLA-4 antibodies in B16F10 tumors.

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Acknowledgements

We are very grateful to Dr. Maya Imbrechts, Gerlanda Vella, Dr. Emanuela Pasciuto, and Prof. Susan Schlenner (KU Leuven, Leuven, Belgium) for their help with the preparation and their advice regarding the design of the flow cytometry experiment described in the current manuscript. We would also like to thank Prof. Damya Laoui and Aleksandar Murgaski (VUB, Brussels, Belgium) for sharing their protocol for tumor dissociation. Finally, we would like to thank all staff members of the Laboratory for Therapeutic and Diagnostic Antibodies (KU Leuven) for their help with the tissue processing for flow cytometry.

Funding

This research is supported by Research Foundation – Flanders (FWO: PhD mandate 1133220N to LJ, and research project G0E2117N to KH and PD) and CELSA (research project CELSA/19/030 to KH and PD).

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Author notes

  1. These authors contributed equally: Kevin Hollevoet, Paul Declerck.

Authors and Affiliations

  1. Laboratory for Therapeutic and Diagnostic Antibodies, KU Leuven – University of Leuven, Leuven, Belgium

    Liesl Jacobs, Kevin Hollevoet & Paul Declerck

  2. Department of Microbiology, Immunology and Transplantation, KU Leuven – University of Leuven, Leuven, Belgium

    Lidia Yshii, Steffie Junius & Adrian Liston

  3. VIB Center for Brain and Disease Research, VIB, Leuven, Belgium

    Lidia Yshii, Steffie Junius & Adrian Liston

  4. PharmAbs – the KU Leuven Antibody Center, KU Leuven – University of Leuven, Leuven, Belgium

    Nick Geukens, Kevin Hollevoet & Paul Declerck

  5. Immunology Programme, Babraham Institute, Cambridge, United Kingdom

    Adrian Liston

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Contributions

LJ, NG, KH, and PD designed the overall study and interpreted the results. LJ performed the experiments and analyzed the data. LY, SJ, and AL contributed to the design and the execution of the flow cytometry experiment and the interpretation of the respective results. LJ wrote the manuscript, which was reviewed and edited by KH and PD. All authors read and approved the manuscript for publication.

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Correspondence to Kevin Hollevoet or Paul Declerck.

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Jacobs, L., Yshii, L., Junius, S. et al. Intratumoral DNA-based delivery of checkpoint-inhibiting antibodies and interleukin 12 triggers T cell infiltration and anti-tumor response. Cancer Gene Ther 29, 984–992 (2022). https://doi.org/10.1038/s41417-021-00403-8

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