Abstract
Antitumor electrochemotherapy is a treatment of solid tumors which combines a cytotoxic nonpermeant drug, like bleomycin, with locally delivered permeabilizing electric pulses (1–3). More generally, a new form of vectorization is achieved by the combination of nonpermeant molecules with intracellular targets and of a physical perturbation that locally permeabilizes the cells. This vectorization does not require chemical, biochemical or biological modifications of the targeted compound, since the modification is performed on the target cells. A very convenient way to transiently permeabilize the cells is the use of appropriate electric pulses (short and intense squarewave electric pulses) that are not cytotoxic by themselves (1). These electric pulses reversibly permeabilize the electropulsed cells. Consequently, they allow increased drug delivery inside cells, particularly in the case of drugs for which the plasma membrane is a barrier that limits their access inside the cell [termed nonpermeant drugs] (4). As illustrated in the various protocols reported in this volume, electrochemotherapy using bleomycin is efficient to eradicate subcutaneously transplanted and spontaneous small tumors in mice and rats as well as experimental internal tumors transplanted in rat brain or in rabbit or rat liver. All the clinical trials (3,5–10) confirm the efficacy of this new therapeutical approach based on an original way to deliver nonpermeant cytotoxic drugs
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References
Mir, L. M., Orlowski, S., Belehradek, J. Jr., and Paoletti, C. (1991) Electrochemotherapy: potentiation of antitumour effect of bleomycin by local electric pulses. Eur. J. Cancer 27, 68–72.
Belehradek, J. Jr., Orlowski, S., Poddevin, B., Paoletti, C., and Mir, L. M. (1991) Electrochemotherapy of spontaneous mammary tumours in mice. Eur. J. Cancer 27, 73–76.
Belehradek, M., Domenge, C., Luboinski, B., Orlowski, S., Belehradek, J. Jr., and Mir L. M. (1993) Electrochemotherapy, a new antitumour treatment: First clinical phase I-II trial. Cancer 72, 3694–3700.
Mir, L. M., Banoun, H., and Paoletti, C. (1988) Introduction of definite amounts of nonpermeant molecules into living cells after electropermeabilization: Direct access to the cytosol. Exp. Cell Res. 175, 15–25.
Domenge, C., Orlowski, S., Luboinski, B., De Baere, T., Schwaab, G., Belehradek, J. Jr., and Mir, L. M. (1996) Antitumor electrochemotherapy: New advances in the clinical protocols. Cancer 77, 956–963.
Heller, R., Jaroszeski, M., Perrott, R., Glass, L. F., Puleo, C., Messina, J. L., Rapaport, D. P., DeConti, R. C., Fenske, N. A., Gilbert, R., Mir, L. M., and Reintgen, D. S. (1996) Phase I/II for the treatment of cutaneous and subcutaneous tumors using electrochemotherapy. Cancer 77, 964–972.
Rudolf, Z., Štabuc, B., Čemažar, M., Miklavčič, D., Vodovnik, L., and Serša, G. (1996) Electrochemotherapy with bleomycin: The first clinical experience in malignant melanoma patients. Radiol. Oncol. 27, 229–235.
Glass, L. F., Fenske, N. A., Jaroszeski, M. J., Perrott, R., Harvey, D. T., Reintgen, D. S., and Heller, R. (1996) Bleomycin-mediated electrochemotherapy of basal cell carcinoma. J. Am. Acad. Dermatol. 34, 82–86.
Glass, L. F., Jaroszeski, M., Gilbert, R., Reintgen, D. S., and Heller, R. (1997) Intralesional bleomycin-mediated electrochemotherapy in 20 patients with basal cell carcinoma. J. Am. Acad. Dermatol. 37, 596–599.
Mir, L. M., Glass, F. L., Serša, G., Teissié, J., Domenge, C., Miklavčič, D., Jaroszeski, M. J., Orlowski, S., Reintgen, D. S., Rudolf, Z., Belehradek, M., Gilbert, R., Rols, M. P., Belehradek, J. Jr, Bachaud, J. M., DeConti, R., Štabuc, B., Čemažar, M., Coninx, P., and Heller, R. (1998) Effective treatment of cutaneous and subcutaneous malignant tumours by electrochemotherapy. Br. J. Cancer 77, 2336–2342.
Zimmermann, U., Pilwat, G., and Riemann, F. (1974) Dielectric breakdown of cell membranes. Biophys. J. 14, 881–899.
Zimmermann, U. (1982) Electric field-mediated fusion and related electrical phenomena. Biochim. Biophys. Acta 694, 227–277.
Neumann, E. (1992) Membrane electroporation and direct gene transfer. Bioelectrochem. Bioenerg. 28, 247–267.
Neumann, E. and Rosenheck, K. (1972) Permeability changes induced by electric impulses in vesicular membranes. J. Membr. Biol. 10, 279–290.
Kinosita, K. Jr. and Tsong, T. Y. (1977) Formation and resealing of pores of controlled sizes in human erythrocyte membrane. Nature 268, 438–441.
Gross, D., Loew, L. M., and Webb, W. W. (1986) Optical imaging of cell membrane potential changes induced by applied electric fields. Biophys. J. 50, 339–348.
Belehradek, J. Jr., Orlowski, S., Ramirez, L. H., Pron, G., Poddevin, B., and Mir, L. M. (1994) Electropermeabilization of cells in tissues assessed by the qualitative and quantitative electroloading of bleomycin. Biochim. Biophys. Acta 1190, 155–163.
Okino, M. and Mohri, H. (1987) Effects of a high voltage electrical impulse and an anticancer drug on in vivo growing tumors. Jpn. J. Cancer Res. (Gann) 78, 1319–1321.
Djuzenova, C. S., Zimmermann, U., Frank, H., Sukhorukov, V. L., Richter, E., and Fuhr, G. (1996) Effect of medium conductivity and composition on the uptake of propidium iodide into electropermeabilized myeloma cells. Biochim. Biophys. Acta 1284, 143–152.
Rols, M-P. and Teissié, J. (1990) Electropermeabilization of mammalian cells. Quantitative analysis of the phenomenon. Biophys. J. 58, 1089–1098.
Poddevin, B., Orlowski, S., Belehradek, J. Jr., and Mior, L. M. (1991) Very high cytotoxicity of bleomycin introduced into the cytosol of cells in culture. Biochem. Pharmacol. 42 (Suppl.), 67-75.
Gehl, J., Sørensen, T. H., Nielsen, K., Raskmark, P., Nielsen, S. L., Skovsgaard, T. and Mir, L. M. (1999) In vivo electroporation of skeletal muscle: threshold, efficacy and relation to electric field distribution. Biochim. Biophys. Acta 1428, 233–240.
Gehl, J. and Mir, L. M. (1999) Determination of optimal parameters for in vivo gene transfer by electroporation, using a rapid in vivo test for cell permeabilization. Biochem. Biophys. Res. Comm. 261, 377–380.
Neumann, E., Schaefer-Ridder, M., Wang, Y., and Hofschneider, P. H. (1982) Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1, 841–845.
Chang, D. C., Gao, P.-Q., and Maxwell, B. L. (1991) High efficiency gene transfection by electroporation using a radio-frequency electric field. Biochim. Biophys. Acta 1092, 153–160.
Gehl, J., Skovsgaard, T., and Mir, L. M. (1998) Enhancement of cytotoxicity by electropermeabilization: An improved method for screening drugs. Anti-Cancer Drugs 9, 319–325.
Takahashi, M., Furukawa, T., Saitoh, H., Aoki, A., Koike, T., Moriyama, Y., and Shibata, A. (1991) Gene transfer into human leukemia cell lines by electroThe Basis of Electrochemotherapy 115 poration: Experience with exponentially decaying and square-wave pulses. Leuk. Res. 15, 507–513.
Okino, M., Tomie, H., Kanesada, H., Marumoto, M., Esato, K., and Suzuki, H. (1992) Optimal electric conditions in electrical impulse chemotherapy. Jpn. J. Cancer Res. (Gann) 83, 1095–1101.
Orlowski, S., Belehradek, J. Jr., Paoletti, C., and Mir, L. M. (1988) Transient electropermeabilization of cells in culture: Increase of the cytotoxicity of anticancer drugs. Biochem. Pharmacol. 37, 4727–4733.
Rols, M-P. and Teissié, J. (1989) Ionic-strength modulation of electrically-induced permeabilization and associated fusion of mammalian cells. Eur. J. Biochem. 179, 109–115.
Serša, G., Čemažar, M., Šemrov, D., and Miklavčič, D. (1996) Changing the electrode orientation improves the efficacy of electrochemotherapy of solid tumors in mice. Bioelectrochem. Bioenerg. 39, 61–66.
Mir, L. M., Tounekti, O., and Orlowski, S. (1996) Bleomycin: Revival of an old drug. Gen. Pharmacol. 27, 745–748.
Pron, G., Belehradek, J. Jr., Orlowski, S., and Mir, L. M. (1994) Involvement of the membrane bleomycin-binding sites in the bleomycin cytotoxicity. Biochem. Pharmacol. 48, 301–310.
Pron, G., Belehradek, J. Jr., and Mir, L. M. (1993) Identification of a plasma membrane protein that specifically binds bleomycin. Biochem. Biophys. Res. Commun. 194, 333–337.
Lazo, J. S., Schisselbauer, J. C., Herring, G. M., and Kennedy, K. A. (1990) Involvement of the cellular vacuolar system with the cytotoxicity of bleomycinlike agents. Cancer Commun. 2, 81–86.
Pron, G., Mahrour, N., Orlowski, S., Tounekti, O., Poddevin, B., Belehradek, J. Jr., and Mir, L. M. (1999) Internalisation of the bleomycin molecules responsible for bleomycin toxicity: A receptor-mediated endocytosis mechanism. Biochem. Pharmacol. 57, 45–56.
Orlowski, S. and Mir, L. M. (1993) Cell electropermeabilization: A new tool for biochemical and pharmacological studies. Biochim. Biophys. Acta 1154, 51–63.
Tounekti, O., Pron, G., Belehradek, J. Jr., and Mir, L. M. (1993) Bleomycin, an apoptosis-mimetic drug that induces two types of cell death depending on the number of molecules internalized. Cancer Res. 53, 5462–5469.
Tounekti, O., Belehradek, J. Jr., and Mir, L. M. (1995) Relationships between DNA fragmentation, chromatin condensation and changes in flow cytometry profiles detected during apoptosis. Exp. Cell Res. 217, 506–516.
Mitchell, M. S. (1988) Combining chemotherapy with biological response modifiers in treatment of cancer. J. Natl. Cancer Inst. 80, 1445–1450.
Morikawa, K., Hosokawa, M., Hamada, J., Xu, Z., and Kobayashi, H. (1986) Possible participation of tumoricidal macrophages in the therapeutic effect of blemycin on a transplantable rat fibrosarcoma. Cancer Res. 46, 684–688.
Morikawa, K., Hosokawa, M., Hamada, J., Sugawara, M., and Kobayashi, H. (1985) Host-mediated therapeutic effects produced by appropriately timed administration of bleomycin on a rat fibrosarcoma. Cancer Res. 45, 1502–1506.
Xu, Z., Hosokawa, M., Morikawa, K., Hatakeyama, M., and Kobayashi, H. (1988) Overcoming suppression of antitumor immune reactivity in tumor-bearing rats by treatment with bleomycin. Cancer Res. 48, 6658–6663.
Abdul Hamied, T. A. and Turk, J. L. (1987) Enhancement of interleukin-2 release in rats by treatment with bleomycin and adriamycin in vivo. Cancer Immunol. Immunother. 25, 245–249.
Heller, R., Jaroszeski, M. J., Perrott, R., Messina, J., and Gilbert, R. (1997) Effective treatment of B16 melanoma by direct delivery of bleomycin using electrochemotherapy. Melanoma Res. 7, 10–18.
Jaroszeski, M. J., Gilbert, R., and Heller, R. (1997) In vivo antitumor effects of electrochemotherapy in a hepatoma model. Biochim. Biophys. Acta 1334, 15–18.
Heller, R., Jaroszeski, M. J., Reintgen, D. S., Puleo, C. A., DeConti, R. C., Gilbert, R. A., and Glass, L. F. (1998) Treatment of cutaneous and subcutaneous tumors with electrochemotherapy using intralesional bleomycin. Cancer 83, 148–157.
Gately, D. F. and Howell, S. B. (1993) Cellular accumulation of the anticancer agent cisplatin: A review. Br. J. Cancer 67, 1171–1176.
Melvik, J. E., Pettersen, E. O., Gordon, P. B., and Seglen, P. O. (1986) Increase in cis-dichlorodiammineplatinum(II) cytotoxicity upon reversible electropermeabilization of the plasma membrane in cultured human NHIK 3025 cells. Eur. J. Cancer Clin. Oncol. 22, 1523–1530.
Serša, G., Čemažar, M., and Miklavčič, D. (1995) Antitumor effectiveness of electrochemotherapy with cis-diamminedichloroplatinum(II) in mice. Cancer Res. 55, 3450–3455.
Čemažar, M., Milačič, R., Miklavčič, D., Dolžan, V., and Serša, G. (1998) Intratumoral cisplatin administration in electrochemotherapy: antitumor effectiveness, sequence dependence and platinum content. Anti-Cancer Drugs 9, 525–530.
Čemažar, M., Miklavčič, D., Ščančar, J., Dolžan, V., Golouh, R., and Serša, G. (1999) Increased platinum accumulation in SA-1 tumour cells after in vivo electrochemotherapy with cisplatin. Br. J. Cancer 79, 1386–1391.
Ramirez, L. H., Orlowski, S., An, D. J., Bindoula, G., Dzodic, R., Ardouin, P., Bognel, C., Belehradek, J. Jr., Munck, J.-N., and Mir, L. M. (1998) Electrochemotherapy on liver tumors in rabbits. Br. J. Cancer 77, 2104–2111.
Serša, G., Beravs, K., Čemažar, M., Miklavčič, D., and Demšar, F. (1998) Contrast enhanced MRI assesment of tumor blood volume after application of electric pulses. Electr. Magnetobiol. 17, 297–304.
Serša, G., Čemažar, M., Parkins, C. S., and Chaplin, D. J. (1999) Tumour blood flow changes induced by application of electric pulses. Eur. J. Cancer 35, 672–677.
Serša, G., Miklavčič, D., Čemažar, M., Belehradek, J. Jr., Jarm, T., and Mir, L. M. (1997) Electrochemotherapy with CDDP on LPB sarcomas: Comparison of the anti-tumor effectiveness in immunocompetent and immunodeficient mice. Bioelectrochem. Bioenerg. 43, 279-283.
Mir, L. M., Orlowski, S., Poddevin, B., and Belehradek, J. Jr. (1992) Electrochemotherapy tumor treatment is improved by interleukin-2 stimulation of host’s defenses. Eur. Cytokine Netw. 3, 331–334.
Serša, G., Čemažar, M., Menart, V., Gaberc-Porekar, V., and Miklavčič, D. (1997) Anti-tumor effectiveness of electrochemotherapy with bleomycin is increased by TNF-α on SA-1 tumors in mice. Cancer Lett. 116, 85–92.
Mir, L. M., Roth, C., Orlowski, S., Quintin-Colonna, F., Fradelizi, D., Belehradek, J. Jr., and Kourilsky, P. (1995) Systemic antitumor effects of electrochemotherapy combined with histoincompatible cells secreting interleukin-2. J. Immunother. 17, 30–38.
Mir, L. M., Roth, C., Orlowski, S., Belehradek, J. Jr., Fradelizi, D., Paoletti, C., and Kourilsky, P. (1992) Potentiation of the antitumoral effect of electrochemotherapy by an immunotherapy with allogeneic cells producing interleukin-2. C. R. Acad. Sci. III 314, 539–544.
Roth, C., Mir, L. M., Cressent, M., Quintin-Colonna, F., Ley, V., Fradelizi, D., and Kourilsky, P. (1992) Inhibition of tumor growth by histoincompatible cells expressing interleukin-2. Int. Immunol. 4, 1429–1436.
Orlowski, S., An, D., Belehradek, J. Jr., and Mir, L. M. (1998) Antimetastatic effects of electrochemotherapy and of histoincompatible interleukin-2-secreting cells in the murine Lewis lung tumor. Anticancer Drugs bd9, 551–556.
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Mir, L.M., Orlowski, S. (2000). The Basis of Electrochemotherapy. In: Jaroszeski, M.J., Heller, R., Gilbert, R. (eds) Electrochemotherapy, Electrogenetherapy, and Transdermal Drug Delivery. Methods in Molecular Medicine, vol 37. Humana Press. https://doi.org/10.1385/1-59259-080-2:99
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DOI: https://doi.org/10.1385/1-59259-080-2:99
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