Abstract
Cell membranes can be transiently permeabilized under the application of electric pulses. This process, called electropermeabilization or electroporation, allows hydrophilic molecules, such as anticancer drugs and DNA, to enter into cells and tissues. The method is nowadays used in clinics to treat cancers. Vaccination and gene therapy are other fields of application of DNA electrotransfer. A description of the mechanisms can be assayed by using different complementary systems with increasing complexities (models of membranes, cells cultivated in 2D and 3D culture named spheroids, and tissues in living mice) and different microscopy tools to visualize the processes from single molecules to entire animals. Single-cell imaging experiments revealed that the uptake of molecules (nucleic acids, antitumor drugs) takes place in well-defined membrane regions and depends on their chemical and physical properties (size, charge). If small molecules freely cross the electropermeabilized membrane and have a free access to the cytoplasm, larger molecules, such as plasmid DNA, face physical barriers (plasma membrane, cytoplasm crowding, nuclear envelope) which reduce transfection efficiency and engender a complex mechanism of transfer. Gene electrotransfer indeed involves different steps that include the initial interaction with the membrane, its crossing, transport within the cytoplasm, and finally gene expression. In vivo, additional very important effects of electric pulses are present such as blood flow modifications. The full knowledge on the way molecules are transported across the electropermeabilized membranes and within tissues is mandatory to improve the efficacy and the safety of the electropermeabilization process both in cell biology and in clinics.
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Andre FM, Mir LM (2010) Nucleic acids electrotransfer in vivo: mechanisms and practical aspects. Curr Gene Ther 10(4):267–280
Beebe SJ, White J et al (2003) Diverse effects of nanosecond pulsed electric fields on cells and tissues. DNA Cell Biol 22(12):785–796
Bellard E, Markelc B et al (2012) Intravital microscopy at the single vessel level brings new insights of vascular modification mechanisms induced by electropermeabilization. J Control Release 163(3):396–403
Cemazar M, Jarm T et al (2010) Cancer electrogene therapy with interleukin-12. Curr Gene Ther 10(4):300–311
Chiarella P, Fazio VM et al (2010) Application of electroporation in DNA vaccination protocols. Curr Gene Ther 10(4):281–286
Chopinet L, Roduit C et al (2013) Destabilization induced by electropermeabilization analyzed by atomic force microscopy. Biochim Biophys Acta 1828(9):2223–2229
Daud AI, DeConti RC et al (2008) Phase I trial of interleukin-12 plasmid electroporation in patients with metastatic melanoma. J Clin Oncol 26(36):5896–5903
Escoffre JM, Rols MP (2012) Electrochemotherapy: progress and prospects. Curr Pharm Des 18:3406–3415
Escoffre JM, Teissie J et al (2010a) Gene transfer: how can the biological barriers be overcome? J Membr Biol 236(1):61–74
Escoffre JM, Kaddur K et al (2010b) In vitro gene transfer by electrosonoporation. Ultrasound Med Biol 36(10):1746–1755
Escoffre JM, Portet T et al (2011) Electromediated formation of DNA complexes with cell membranes and its consequences for gene delivery. Biochim Biophys Acta 1808(6):1538–1543
Escoffre JM, Bellard E et al (2014a) Membrane disorder and phospholipid scrambling in electropermeabilized and viable cells. Biochim Biophys Acta 1838(7):1701–1709
Escoffre JM, Hubert M et al (2014b) Evidence for electro-induced membrane defects assessed by lateral mobility measurement of a GPi anchored protein. Eur Biophys J 43:277–286
Faurie C, Rebersek M et al (2010) Electro-mediated gene transfer and expression are controlled by the life-time of DNA/membrane complex formation. J Gene Med 12(1):117–125
Frandsen SK, Gibot L et al (2015) Calcium electroporation: evidence for differential effects in normal and malignant cell lines, evaluated in a 3D spheroid model. PLoS One 10(12):e0144028
Gehl J, Skovsgaard T et al (2002) Vascular reactions to in vivo electroporation: characterization and consequences for drug and gene delivery. Biochim Biophys Acta 1569(1–3):51–58
Gibot L, Rols MP (2013) Progress and prospects: the use of 3D spheroid model as a relevant way to study and optimize DNA electrotransfer. Curr Gene Ther 13(3):175–181
Gibot L, Wasungu L et al (2013) Antitumor drug delivery in multicellular spheroids by electropermeabilization. J Control Release 167(2):138–147
Golzio M, Teissie J (2014) siRNA delivery via electropulsation: a review of the basic processes. Methods Mol Biol 1121:81–98
Golzio M, Teissie J et al (2002) Direct visualization at the single-cell level of electrically mediated gene delivery. Proc Natl Acad Sci U S A 99(3):1292–1297
Heller LC, Heller R (2010) Electroporation gene therapy preclinical and clinical trials for melanoma. Curr Gene Ther 10(4):312–317
Heller R, Heller LC (2015) Gene electrotransfer clinical trials. Adv Genet 89:235–262
Kamensek U, Rols MP et al (2016) Visualization of nonspecific antitumor effectiveness and vascular effects of gene electro-transfer to tumors. Curr Gene Ther 16(2):90–97
Madi M, Rols MP et al (2015) Efficient in vitro electropermeabilization of reconstructed human dermal tissue. J Membr Biol 248:903–908
Marrero B, Heller R (2012) The use of an in vitro 3D melanoma model to predict in vivo plasmid transfection using electroporation. Biomaterials 33(10):3036–3046
Mauroy C, Castagnos P et al (2012a) Interaction between GUVs and catanionic nanocontainers: new insight into spontaneous membrane fusion. Chem Commun (Camb) 48(53):6648–6650
Mauroy C, Portet T et al (2012b) Giant lipid vesicles under electric field pulses assessed by non invasive imaging. Bioelectrochemistry 87:253–259
Mir LM, Glass LF et al (1998) Effective treatment of cutaneous and subcutaneous malignant tumours by electrochemotherapy. Br J Cancer 77(12):2336–2342
Neumann E, Schaefer-Ridder M et al (1982) Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J 1(7):841–845
Paganin-Gioanni A, Bellard E et al (2011) Direct visualization at the single-cell level of siRNA electrotransfer into cancer cells. Proc Natl Acad Sci U S A 108(26):10443–10447
Portet T, Camps i Febrer F et al (2009) Visualization of membrane loss during the shrinkage of giant vesicles under electropulsation. Biophys J 96(10):4109–4121
Portet T, Favard C et al (2011) Insights into the mechanisms of electromediated gene delivery and application to the loading of giant vesicles with negatively charged macromolecules. Soft Matter 7(8):3872–3881
Ravi M, Ramesh A et al (2016) Contributions of 3D cell cultures for cancer research. J Cell Physiol 232(10): 2679–2697
Rols MP, Delteil C et al (1998) In vivo electrically mediated protein and gene transfer in murine melanoma. Nat Biotechnol 16(2):168–171
Rosazza C, Escoffre JM et al (2011) The actin cytoskeleton has an active role in the electrotransfer of plasmid DNA in mammalian cells. Mol Ther 19(5):913–921
Rosazza C, Buntz A et al (2013) Intracellular tracking of single plasmid DNA-particles after delivery by electroporation. Mol Ther 21:2217–2226
Rosazza C, Meglic SH et al (2016) Gene electrotransfer: a mechanistic perspective. Curr Gene Ther 16(2):98–129
Sersa G, Cemazar M et al (1999) Tumor blood flow modifying effect of electrochemotherapy with bleomycin. Anticancer Res 19(5B):4017–4022
Sersa G, Teissie J et al (2015) Electrochemotherapy of tumors as in situ vaccination boosted by immunogene electrotransfer. Cancer Immunol Immun 64:1315–1327
Sutherland RM (1988) Cell and environment interactions in tumor microregions: the multicell spheroid model. Science 240(4849):177–184
Teissie J, Golzio M et al (2005) Mechanisms of cell membrane electropermeabilization: a minireview of our present (lack of ?) knowledge. Biochim Biophys Acta 1724(3):270–280
Yarmush ML, Golberg A et al (2014) Electroporation-based technologies for medicine: principles, applications, and challenges. Annu Rev Biomed Eng 16:295–320
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Gibot, L., Golzio, M., Rols, MP. (2017). How Imaging Membrane and Cell Processes Involved in Electropermeabilization Can Improve Its Development in Cell Biology and in Clinics. In: Kulbacka, J., Satkauskas, S. (eds) Transport Across Natural and Modified Biological Membranes and its Implications in Physiology and Therapy. Advances in Anatomy, Embryology and Cell Biology, vol 227. Springer, Cham. https://doi.org/10.1007/978-3-319-56895-9_7
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DOI: https://doi.org/10.1007/978-3-319-56895-9_7
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