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Antitumor effect of antibiotic resistance gene-free plasmids encoding interleukin-12 in canine melanoma model

Cancer Gene Therapy volume 25, pages 260–273 (2018)Cite this article

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Abstract

The electrotransfer of interleukin-12 (IL-12) has been demonstrated as an efficient and safe treatment for tumors in veterinary oncology. However, the plasmids used encode human or feline IL-12 and harbor the gene for antibiotic resistance. Therefore, our aim was to construct plasmids encoding canine IL-12 without the antibiotic resistance genes driven by two different promoters: constitutive and fibroblast-specific. The results obtained in vitro in different cell lines showed that following gene electrotransfer, the newly constructed plasmids had cytotoxicity and expression profiles comparable to plasmids with antibiotic resistance genes. Additionally, in vivo studies showed a statistically significant prolonged tumor growth delay of CMeC-1 tumors compared to control vehicle-treated mice after intratumoral gene electrotransfer. Besides the higher gene expression obtained by plasmids with constitutive promoters, the main difference between both plasmids was in the distribution of the transgene expression. Namely, after gene electrotransfer, plasmids with constitutive promoters showed an increase of serum IL-12, whereas the gene expression of IL-12, encoded by plasmids with fibroblast-specific promoters, was restricted to the tumor. Furthermore, after the gene electrotransfer of plasmids with constitutive promoters, granzyme B-positive cells were detected in the tumor and spleen, indicating a systemic effect of the therapy. Therefore, plasmids with different promoters present valuable tools for focused therapy with local or systemic effects. The results of the present study demonstrated that plasmids encoding canine IL-12 under constitutive and fibroblast-specific promoters without the gene for antibiotic resistance provide feasible tools for controlled gene delivery that could be used for the treatment of client-owned dogs.

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Acknowledgements

This work was financially supported through grants from the state budget of the Slovenian Research Agency (program no. P3-0003, projects no. J3-6796 and J3-4259). The research was conducted in the scope of LEA EBAM (French-Slovenian European Associated Laboratory: Pulsed Electric Fields Applications in Biology and Medicine) and reflects the networking efforts within COST TD1104 Action. Linguistic revision was done by American Journal Experts.

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Authors and Affiliations

  1. Department of Experimental Oncology, Institute of Oncology Ljubljana, Zaloška 2, 1000, Ljubljana, Slovenia

    Ursa Lampreht Tratar, Spela Kos, Urska Kamensek, Gregor Sersa & Maja Cemazar

  2. Department of Pathology, Institute of Oncology Ljubljana, Zaloška 2, 1000, Ljubljana, Slovenia

    Maja Ota

  3. Clinic for Surgery and Small Animals, University of Ljubljana, Veterinary Faculty, Cesta v mestni log 47, 1000, Ljubljana, Slovenia

    Natasa Tozon

  4. Faculty of Health Sciences, University of Ljubljana, Zdravstvena pot 5, 1000, Ljubljana, Slovenia

    Gregor Sersa

  5. Faculty of Health Sciences, University of Primorska, Polje 42, Izola, 6310, Slovenia

    Maja Cemazar

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Correspondence to Maja Cemazar.

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Lampreht Tratar, U., Kos, S., Kamensek, U. et al. Antitumor effect of antibiotic resistance gene-free plasmids encoding interleukin-12 in canine melanoma model. Cancer Gene Ther 25, 260–273 (2018). https://doi.org/10.1038/s41417-018-0014-5

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