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Particulate mediators of the bystander effect linked to suicide and interferon-β transgene expression in melanoma cells

Gene Therapy volume 28, pages 38–55 (2021)Cite this article

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Abstract

In the context of comparative oncology, melanoma cells derived from companion animal tumors are good models for optimizing and predicting their in vivo response to therapeutic strategies. Here, we report that human, canine, and feline melanoma cells driven to death by bleomycin, interferon-β gene, or herpes simplex virus thymidine kinase/ganciclovir suicide gene (SG) treatment significantly increased their internal granularity. This fact correlated with the release of a heterogeneous collection of nano- and micro-sized granules as revealed by transmission electron microscopy. While killing lipofected cells, the expressed transgenes and their derived products were incorporated into these granules that were isolated by differential centrifugation. These particulate factors (PFs) were able to transfer, in a dose- and time-dependent manner, appreciable levels of therapeutic genes, related proteins, and drugs. Thus, when recipient cells of SG-carrying PFs were exposed to ganciclovir, this prodrug was efficiently activated, eliminating them. These PFs kept the functionality of their cargo, even after being subjected to adverse conditions, such as the presence of DNase, freezing, or heating. Since our in vitro system did not include any of the immune mechanisms that could provide additional antitumor activity, the chemo-gene treatments amplified by these delivery bags of therapeutic agents offer a great clinical potential.

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Fig. 1: Induction of  intracellular granularity by antitumoral agents.
Fig. 2: Correlations between intracellular granularity and extracellular granules.
Fig. 3: Quantification and cytotoxic effects of the particulate factors released by suicide gene.
Fig. 4: Plasmidic DNAs and their translated proteins carried by particulate factors.
Fig. 5: Direct or particulate factors mediated cytotoxic effects of the treatments.
Fig. 6: Dose, incubation time and heparin effects on particulate factors mediated cytotoxicity.
Fig. 7: Direct or particulate factors mediated cytotoxic effects of treatments.

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Acknowledgements

We thank Juan M. Cardini and Graciela B. Zenobi for technical assistance. This study was partially supported by grants from ANPCYT/FONCYT (PICT2014-1652) and CONICET (PIP 11220150100885). GCG and LMEF are investigators, and LA, CF, and MFA fellows of the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET, Argentina).

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  1. Unidad de Transferencia Genética, Instituto de Oncología “Ángel H. Roffo”, Universidad de Buenos Aires, Buenos Aires, Argentina

    Lucrecia Agnetti, Chiara Fondello, María Florencia Arbe, Gerardo C. Glikin & Liliana M. E. Finocchiaro

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Correspondence to Liliana M. E. Finocchiaro.

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Agnetti, L., Fondello, C., Arbe, M. et al. Particulate mediators of the bystander effect linked to suicide and interferon-β transgene expression in melanoma cells. Gene Ther 28, 38–55 (2021). https://doi.org/10.1038/s41434-020-0136-x

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