Abstract
Purpose. The aim of this work was to investigate the cellular binding and processing of polyethylene glycol-stabilized epidermal growth factor (EGF) liposomes. The liposomes were actively loaded with water-soluble boronated acridine (WSA), primarily developed for boron neutron capture therapy.
Methods. The uptake, internalization, and retention of EGF-liposome conjugates were studied in two cultured monolayer cell-lines, A-431 and U-343, with regard to the nuclide-label on the targeting agent, the carrier, and the load. The subcellular localization of WSA was studied using confocal microscopy.
Results. We found that the liposome complex was internalized after specific binding to the EGF receptor. After internalization in the tumor cells, WSA was distributed mainly in the cytoplasm and was shown to have long cellular retention, with 80% of the boron remaining after 48 h.
Conclusions. The long retention of the compound and the cellular boron concentration reached makes these targeted liposomes interesting for further development toward boron neutron capture therapy.
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REFERENCES
G. Gregoriadis, E. J. Wills, C. P. Swain, and A. S. Tavill. Drug-carrier potential of liposomes in cancer chemotherapy. Lancet 1:1313-1316 (1974).
D. Papahadjopoulos, T. M. Allen, A. Gabizon, E. Mayhew, K. Matthay, S. K. Huang, K. D. Lee, M. C. Woodle, D. D. Lasic, C. Redemann, and F. J. Martin. Sterically stabilized liposomes: improvements in pharmacokinetics and antitumor therapeutic efficacy. Proc. Natl. Acad. Sci. USA 88:11460-11464 (1991).
A. A. Gabizon. Selective tumor localization and improved therapeutic index of anthracyclines encapsulated in long-circulating liposomes. Cancer Res. 52:891-896 (1992).
A. L. Klibanov, K. Maruyama, V. P. Torchilin, and L. Huang. Amphipathic polyethyleneglycols effectively prolong the circulation time of liposomes. FEBS Lett. 268:235-237 (1990).
I. A. Bakker-Woudenberg, A. F. Lokerse, M. T. ten Kate, J. W. Mouton, M. C. Woodle, and G. Storm. Liposomes with prolonged blood circulation and selective localization in Klebsiella pneumoniae-infected lung tissue. J. Infect. Dis. 168:164-171 (1993).
T. M. Allen, E. Brandeis, C. B. Hansen, G. Y. Kao, and S. Zalipsky. A new strategy for attachment of antibodies to sterically stabilized liposomes resulting in efficient targeting to cancer cells. Biochim. Biophys. Acta 1237:99-108 (1995).
J. Carlsson, L. Gedda, C. Gronvik, T. Hartman, A. Lindstrom, P. Lindstrom, H. Lundqvist, A. Lovqvist, J. Malmqvist, P. Olsson, S. Mayfield, J. P. Vaughn, T. E. Kute, W. A. Denny, R. L. Gutman, G. Peacock, and D. R. Lu. Strategy for boron neutron capture therapy against tumor cells with over-expression of the epidermal growth factor-receptor. Int. J. Radiat. Oncol. Biol. Phys. 30:105-115 (1994).
Y. Ishii, Y. Aramaki, T. Hara, S. Tsuchiya, and T. Fuwa. Preparation of EGF labeled liposomes and their uptake by hepatocytes. Biochem. Biophys. Res. Commun. 160:732-736 (1989).
A. Kikuchi, S. Sugaya, H. Ueda, K. Tanaka, Y. Aramaki, T. Hara, H. Arima, S. Tsuchiya, and T. Fuwa. Efficient gene transfer to EGF receptor overexpressing cancer cells by means of EGF-labeled cationic liposomes. Biochem. Biophys. Res. Commun. 227:666-671 (1996).
D. C. Drummond, K. Hong, J. W. Park, C. C. Benz, and D. B. Kirpotin. Liposome targeting to tumors using vitamin and growth factor receptors. Vitam. Horm. 60:285-332 (2000).
J. W. Park, D. B. Kirpotin, K. Hong, R. Shalaby, Y. Shao, U. B. Nielsen, J. D. Marks, D. Papahadjopoulos, and C. C. Benz. Tumor targeting using anti-her2 immunoliposomes. J. Control. Release 74:95-113 (2001).
D. E. Lopes de Menezes, M. J. Kirchmeier, J.-F. Gagne, L. M. Pilarski, and T. M. Allen. Cellular trafficking and cytotoxicity of anti-CD19-targeted liposomal doxorubicin in B lymphoma cells. J. Liposome Res. 9:199-228 (1999).
A. Gabizon, A. T. Horowitz, D. Goren, D. Tzemach, F. Mandelbaum-Shavit, M. M. Qazen, and S. Zalipsky. Targeting folate receptor with folate linked to extremities of poly(ethylene glycol)-grafted liposomes: in vitro studies. Bioconjug. Chem. 10:289-298 (1999).
T. Hartman, H. Lundqvist, J. E. Westlin, and J. Carlsson. Radiation doses to the cell nucleus in single cells and cells in micrometastases in targeted therapy with (131)I labeled ligands or antibodies. Int. J. Radiat. Oncol. Biol. Phys. 46:1025-1036 (2000).
H. Ghaneolhosseini, W. Tjarks, and S. Sjöberg. Synthesis of novel boronated acridines-and spermidines as possible agents for BNCT. Tetrahedon 54:3877-3884 (1998).
R. F. Barth, A. H. Soloway, and R. M. Brugger. Boron neutron capture therapy of brain tumors: past history, current status and future potential. Cancer Invest. 14:534-550 (1996).
M. Johnsson, N. Bergstrand, and K. Edwards. Optimization of drug loading procedures and characterization of liposomal formulations of two novel agents intended for boron neutron capture therapy (BNCT). J. Liposome Res. 9:53-79 (1999).
B. Westermark, A. Magnusson, and C.-H. Heldin. Effect of epidermal growth factor on membrane motility and cell locomotion in cultures of human clonal glioma cells. J. Neurol. Res. 8:491-507 (1982).
H. Haigler, J. F. Ash, S. J. Singer, S. Cohen, F. M. Yakes, W. Chinratanalab, C. A. Ritter, W. King, S. Seelig, and C. L. Arteaga. Visualization by fluorescence of the binding and internalization of epidermal growth factor in human carcinoma cells A-431. Proc. Natl. Acad. Sci. USA 75:3317-3321 (1978).
E. Bohl Kullberg, N. Bergstrand, J. Carlsson, K. Edwards, M. Johnsson, S. Sjoberg, and L. Gedda. Development of EGF-conjugated liposomes for targeted delivery of boronated DNA-binding agents. Bioconjug. Chem. 13:737-743 (2002).
D. D. Lasic, B. Ceh, M. C. Stuart, L. Guo, P. M. Frederik, and Y. Barenholz. Transmembrane gradient driven phase transitions within vesicles: lessons for drug delivery. Biochim. Biophys. Acta 1239:145-156 (1995).
H. T. Haigler, F. R. Maxfield, M. C. Willingham, and I. Pastan. Dansylcadaverine inhibits internalization of 125I-Epidermal Growth Factor in BALB 3T3 cells. J. Biol. Chem. 255:1239-1241 (1980).
P. Lindström, P. Olsson, J. Malmqvist, J. Pettersson, P. Lemmen, B. Werner, S. Sjöberg, å. Olin, and J. Carlsson. New carborane-based compounds for boron neutron capture therapy: binding and toxicity of ANC-1, DAC-1 and B-Et-11-OMe in cultured human glioma and mouse melanoma cells. Anticancer Drugs 5:43-52 (1994).
P. Olsson, A. Lindstrom, and J. Carlsson. Internalization and excretion of epidermal growth factor-dextran-associated radioactivity in cultured human squamous-carcinoma cells. Int. J. Cancer 56:529-537 (1994).
D. Goren, A. T. Horowitz, D. Tzemach, M. Tarshish, S. Zalipsky, and A. Gabizon. Nuclear delivery of doxorubicin via folate-targeted liposomes with bypass of multidrug-resistance efflux pump. Clin. Cancer Res. 6:1949-1957 (2000).
D. E. Lopes de Menezes, L. M. Pilarski, A. R. Belch, and T. M. Allen. Selective targeting of immunoliposomal doxorubicin against human multiple myeloma in vitro and ex vivo. Biochim. Biophys. Acta 1466:205-220 (2000).
A. Sorkin and C. M. Waters. Endocytosis of growth factor receptors. Bioessays 15:375-382 (1993).
E. Mastrobattista, G. A. Koning, and G. Storm. Immunoliposomes for the targeted delivery of antitumor drugs. Adv. Drug Deliv. Rev. 40:103-127 (1999).
V. A. Slepushkin, S. Simoes, P. Dazin, M. S. Newman, L. S. Guo, M. C. Pedroso de Lima, and N. Duzgunes. Sterically stabilized pH-sensitive liposomes. Intracellular delivery of aqueous contents and prolonged circulation in vivo. J. Biol. Chem. 272:2382-2388 (1997).
D. C. Drummond, M. Zignani, and J. Leroux. Current status of pH-sensitive liposomes in drug delivery. Prog. Lipid Res. 39:409-460 (2000).
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Kullberg, E.B., Nestor, M. & Gedda, L. Tumor-Cell Targeted Epidermal Growth Factor Liposomes Loaded with Boronated Acridine: Uptake and Processing. Pharm Res 20, 229–236 (2003). https://doi.org/10.1023/A:1022223204460
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DOI: https://doi.org/10.1023/A:1022223204460