Abstract
Gene electrotransfer (GET) is a reliable and effective physical method for in vivo delivery of plasmid DNA (pDNA). Several preclinical and clinical studies have utilized GET to deliver plasmids encoding immune stimulating genes for treatment of melanoma and other tumor types. Intratumor delivery of plasmids encoding cytokines directly to tumors can induce not only a local immune response, but a systemic one as well. To obtain an effective immune response, it is critical to achieve the appropriate expression pattern of the delivered transgene. Expression pattern (levels and kinetics) can be modified by manipulating the electrotransfer parameters. These parameters include the tissue target and the electric pulse parameters of pulse width, electric field, and pulse number. We have found that to induce a robust immune response, we needed only low to moderately elevated expression levels compared to controls. When developing a therapeutic protocol, it is important to establish what expression profile will enable the appropriate response. In this chapter we describe how to determine the appropriate GET protocol to achieve the expression profile that can result in the desired clinical response.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Heller LC (2016) Principles of electroporation for gene therapy. In: Miklavcic D (ed) Handbook of electroporation, 1st edn. Springer International Publishing, Cham
Shirley SA, Heller R, Heller LC (2013) Electroporation gene therapy. In: Lattime EC, Gerson SL (eds) Gene therapy of Cancer: translational approaches from preclinical studies to clinical implementation, 3rd edn. Academic Press, San Diego
Young JL, Dean DA (2015) Electroporation-mediated gene delivery. Adv Genet 89:49–88
Vannucci L, Lai M, Chiuppesi F et al (2013) Viral vectors: a look back and ahead on gene transfer technology. New Microbiol 36(1):1–22
Wirth T, Parker N, Yla-Herttuala S (2013) History of gene therapy. Gene 525(2):162–169
Kaufmann KB, Buning H, Galy A et al (2013) Gene therapy on the move. EMBO Mol Med 5(11):1642–1661
Heller R, Heller LC (2015) Gene electrotransfer clinical trials. Adv Genet 89:235–262
Heller LC, Heller R (2010) Electroporation gene therapy preclinical and clinical trials for melanoma. Curr Gene Ther 10(4):312–317
Gothelf A, Gehl J (2010) Gene electrotransfer to skin; review of existing literature and clinical perspectives. Curr Gene Ther 10(4):287–299
Sokolowska E, Blachnio-Zabielska AU (2019) A critical review of electroporation as a plasmid delivery system in mouse skeletal muscle. Int J Mol Sci 20(11):2776
Marshall WG Jr, Boone BA, Burgos JD et al (2010) Electroporation-mediated delivery of a naked DNA plasmid expressing VEGF to the porcine heart enhances protein expression. Gene Ther 17(3):419–423
Sersa G, Teissie J, Cemazar M et al (2015) Electrochemotherapy of tumors as in situ vaccination boosted by immunogene electrotransfer. Cancer Immunol Immunother 64(10):1315–1327
Savarin M, Kamensek U, Cemazar M et al (2017) Electrotransfer of plasmid DNA radiosensitizes B16F10 tumors through activation of immune response. Radiol Oncol 51(1):30–39
Milevoj N, Tratar UL, Nemec A et al (2019) A combination of electrochemotherapy, gene electrotransfer of plasmid encoding canine IL-12 and cytoreductive surgery in the treatment of canine oral malignant melanoma. Res Vet Sci 122:40–49
Cemazar M, Golzio M, Sersa G et al (2009) Control by pulse parameters of DNA electrotransfer into solid tumors in mice. Gene Ther 16(5):635–644
Shirley SA, Lundberg CG, Li F et al (2015) Controlled gene delivery can enhance therapeutic outcome for cancer immune therapy for melanoma. Curr Gene Ther 15(1):32–43
Soden D, Larkin J, Collins C et al (2004) The development of novel flexible electrode arrays for the electrochemotherapy of solid tumour tissue. (Potential for endoscopic treatment of inaccessible cancers). Paper presented at the annual international conference of the IEEE engineering in medicine and biology society, 5:3547–3550
Soden DM, Larkin JO, Collins CG et al (2006) Successful application of targeted electrochemotherapy using novel flexible electrodes. Cancer Lett 232(2):300–310
Agerholm-Larsen B, Iversen HK, Ibsen P et al (2011) Preclinical validation of electrochemotherapy as an effective treatment for brain tumors. Cancer Res 71(11):3753–3762
Lucas ML, Heller L, Coppola D et al (2002) IL-12 plasmid delivery by in vivo electroporation for the successful treatment of established subcutaneous B16. F10 Melanoma Mol Ther 5(6):668–675
Lucas ML, Heller R (2003) IL-12 gene therapy using an electrically mediated nonviral approach reduces metastatic growth of melanoma. DNA Cell Biol 22(12):755–763
Shi G, Edelblute C, Arpag S et al (2018) IL-12 Gene electrotransfer triggers a change in immune response within mouse tumors. Cancers 10(12):498. https://doi.org/10.3390/cancers10120498
Marrero B, Shirley S, Heller R (2014) Delivery of interleukin-15 to B16 melanoma by electroporation leads to tumor regression and long-term survival. Technol Cancer Res Treat 13(6):551–560
Daud AI, DeConti RC, Andrews S et al (2008) Phase I trial of interleukin-12 plasmid electroporation in patients with metastatic melanoma. J Clin Oncol 26(36):5896–5903
Algazi A, Bhatia S, Agarwala S et al (2020) Intratumoral delivery of tavokinogene telseplasmid yields systemic immune responses in metastatic melanoma patients. Ann Oncol 31(4):532–540
Greaney SK, Algazi AP, Tsai KK et al (2020) Intratumoral plasmid IL12 electroporation therapy in patients with advanced melanoma induces systemic and intratumoral T-cell responses. Cancer Immunol Res 8(2):246–254
Algazi AP, Twitty CG, Tsai KK et al (2020) Phase II trial of IL-12 plasmid transfection and PD-1 blockade in immunologically quiescent melanoma. Clin Cancer Res. https://doi.org/10.1158/1078-0432.CCR-19-2217
Gilbert R, Jaroszeski MJ, Heller R (1997) Novel electrode designs for electrochemotherapy. Biochem Biophys Acta 1334:9–14
Heller R, Cruz Y, Heller LC et al (2010) Electrically mediated delivery of plasmid DNA to the skin, using a multielectrode array. Hum Gene Ther 21(3):357–362
Heller L, Coppola D (2002) Electrically mediated delivery of vector plasmid DNA elicits an antitumor effect. Gene Ther 9(19):1321–1325
Heller LC, Cruz YL, Ferraro B et al (2010) Plasmid injection and application of electric pulses alter endogenous mRNA and protein expression in B16. F10 mouse melanomas. Cancer Gene Ther 17(12):864–871
Heller L, Todorovic V, Cemazar M (2013) Electrotransfer of single-stranded or double-stranded DNA induces complete regression of palpable B16. F10 mouse melanomas. Cancer Gene Ther 20(12):695–700
Znidar K, Bosnjak M, Cemazar M et al (2016) Cytosolic DNA sensor upregulation accompanies DNA electrotransfer in B16.F10 melanoma cells. Mol Ther Nucleic Acids 5(6):e322. https://doi.org/10.1038/mtna.2016.34
Znidar K, Bosnjak M, Semenova N et al (2018) Tumor cell death after electrotransfer of plasmid DNA is associated with cytosolic DNA sensor upregulation. Oncotarget 9(27):18665–18681
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Heller, R., Shi, G. (2021). Controlled Delivery of Plasmid DNA to Melanoma Tumors by Gene Electrotransfer. In: Hargadon, K.M. (eds) Melanoma. Methods in Molecular Biology, vol 2265. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1205-7_43
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1205-7_43
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1204-0
Online ISBN: 978-1-0716-1205-7
eBook Packages: Springer Protocols