Cutaneous antiangiogenesis by gene electrotransfer
(2011)
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- (en) Despite the discovery of novel inhibitors of tumor angiogenesis, protein-based antiangiogenic cancer therapy suffers some limitations that antiangiogenic gene therapy could overcome. We investigated the effect on tumor growth and metastasis of three antiangiogenic plasmids administered by two intratumoral electrotransfers. Plasmids encoding recombinant disintegrin domain of ADAM-15 (RDD), thrombospondin 1 (TSP-1) and the soluble isoform of the VEGF receptor 1 (sFlt-1) were injected into B16F10 melanoma-bearing C57BL/6 mice followed by electroporation. Tumor volume was measured daily using a digital caliper. Metastasis was monitored by in vivo bioluminescence after surgical removal of the primary luciferase-encoding tumor 5 days after intratumoral electrotransfer. Markers of vascularization and cell proliferation were quantified by immunohistochemistry. All the plasmids induced a significant inhibition of tumor growth, doubling of mean survival time and long term survivors (~40% vs 0% in control). When the tumor was removed by surgery after intratumoral plasmid electrotransfer, a significant decrease in tumor metastasis was observed leading to long term tumor-free survival especially after treatment with pRDD plasmid (84% vs 0% in control). Unlike pTSP-1 and psFlt-1, pRDD significantly decreased cell proliferation in B16F10 primary tumors which express αvβ3 and α5β1 integrins. No effect of antiangiogenic plasmid electrotransfer on normal skin blood flow was detected. The intratumoral electrotransfer of the plasmid encoding RDD is a promising method for the treatment of melanoma due to its direct antitumoral activity combined with angiogenesis suppression. The association of microneedles with electric pulses causing electroporation could result in an efficient and less painful delivery of drugs and DNA into the skin. Hollow conductive microneedles were used for i) needle-free intradermal injection ii) electric pulse application in order to achieve electric field in the superficial layers of the skin, sufficient for electroporation. Microneedle array was used in combination with a vibratory inserter to disrupt the stratum corneum, thus piercing the skin. Effective injection of proteins into the skin was achieved, resulting in an immune response directed to the model antigen ovalbumin. However, when used both as microneedles to inject and as electrodes to apply the electric pulses, the set-up showed several limitations for DNA electrotransfer. This could be due to the distribution of the electric field in the skin as shown by numerical calculations and/or the low dose of DNA injected. Further investigation of these parameters is needed in order to optimize minimally invasive DNA electrotransfer in the skin.
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Daugimont, L. (2011). Cutaneous antiangiogenesis by gene electrotransfer. https://hdl.handle.net/2078.5/152889