Abstract
A number of drugs are regarded as possessing local activity because their effects take place at an extremely short distance from their location site in the cell. The response of different cellular compartments to these effects is different. Such substances as photosensitizers (PSs), which are used in photodynamic cancer therapy, should be targeted to the cell compartments where their effect is the most pronounced. This study describes the construction and properties of the chimeric modular recombinant transporters (MRTs) expressed in Escherichia coli and used for PS targeting. These constructs include (1) the α-melanocyte-stimulating hormone as a ligand module, which is internalized by the target cells (mouse melanoma); (2) the optimized SV40 large T-antigen nuclear localization signal; (3) the hemoglobin-like protein from E. coli as a carrier module; (4) the endosomolytic module, the translocation domain of the diphtheria toxin. These MRTs were used for PS targeting to the mouse melanoma cell nuclei, the most PS-damaged intracellular compartment, which resulted in a PS photocytotoxic effect increase of several orders of magnitude. In our opinion, MRTs, which target locally active drugs into the desired cell compartment and thereby enhance the drug response, represent a new generation of the pharmacological agents.
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Sobolev, A.S., Jans, D.A., and Rosenkranz, A.A., Targeted Intracellular Delivery of Photosensitizers, Progr. Biophys. Mol. Biol., 2000, vol. 73, pp. 51-90.
Rosenkranz, A.A., Jans, D.A., and Sobolev, A.S., Targeted Intracellular Delivery of Photosensitizers to Enhance Photodynamic Efficiency, Immunol. Cell Biol., 2000, vol. 78, pp. 452-464.
Akhlynina, T.V., Jans, D.A., Statsyuk, N.V., et al., Adenovirus Synergize with Nuclear Localization Signals to Enhance the Nuclear Delivery and Photodynamic Action of Internalizable Conjugates Containing Chlorin e 6, Int. J. Cancer, 1999, vol. 81, pp. 734-740.
Akhlynina, T.V., Jans, D.A., Rosenkranz, A.A., et al., Nuclear Targeting of Chlorin e 6 Enhances Its Photosensitizing Activity, J. Biol. Chem., 1997, vol. 272, pp. 20328-20331.
Bisland, S.K., Singh, D., and Gariepy, J., Potentiation of Chlorin e 6 Photodynamic Activity in Vitro with Peptide-Based Intracellular Vehicles, Bioconjugate Chem., 1999, vol. 10, pp. 982-992.
Jiang, J., Sharma, S.D., Fink, J.L., et al., Melanotropic Peptide Receptors: Membrane Markers of Human Melanoma Cells., Exp. Dermatol., 1996, vol. 5, pp. 325-333.
Funasaka, Y., Sato, H., Chakraborty, A.K., et al., Expression of Proopiomelanocortin, Corticotropin-Releasing Hormone (CRH), and CRH Receptor in Melanoma Cells, Nevus Cells, and Normal Human Melanocytes, J. Invest. Dermatol. Symp. Proc., 1999, vol. 4, pp. 105-109.
Chen, J., Cheng, Z., Miao, Y., et al., A Melanocyte-Stimulating Hormone Peptide Analogs Labeled with Technetium-99m and Indium-11 for Malignant Melanoma Targeting, Cancer, 2002, vol. 94, pp. 1196-1201.
Mendelsohn, J. and Baselga, J., The EGF Family Receptors as Targets for Cancer Therapy, Oncogene, 2000, vol. 19, pp. 6550-6565.
Van Eijck, C.H., de Jong, M., Breeman, W.A., et al., Somatostatin Receptor Imaging and Therapy of Pancreatic Endocrine Tumors, Ann. Oncol., 1999, vol. 10, no. 4, pp. 177-181.
De Jong, M., Valkema, R., Jamar, F., et al., Somatostatin Receptor-Targeted Radionuclide Therapy of Tumors: Preclinical and Clinical Findings, Semin. Nucl. Med., 2002, vol. 32, pp. 133-140.
Plank, C., Oberhauser, B., Mechtler, K., et al., The Influence of Endosome-Disruptive Peptides on Gene Transfer Using Synthetic Virus-Like Gene Transfer Systems, J. Biol. Chem., 1994, vol. 269, pp. 12 918-12 924.
Siegrist, W. and Eberle, A., In Situ Melanin Assay for MSH Using Mouse B16 Melanoma Cells in Culture, Anal. Biochem., 1986, vol. 159, pp.191-197.
O'Hare, K.B., Duncan, R., Strohalm, J., et al., Polymeric Drug-Carriers Containing Doxorubicin and Melanocyte-Stimulating Hormone: In Vitro and In Vivo Evaluation against Murine Melanoma, J. Drug Target., 1993, vol. 1, pp. 217-229.
Wen, Z., Tao, X., Lakkis, F., et al., Diphtheria Toxin-Related ?-Melanocyte-Stimulating Hormone Fusion Toxin: Internal In-Frame Deletion from Thr387 to His485 Results in the Formation of a Highly Potent Fusion Toxin Which Is Resistant to Proteolytic Degradation, J. Biol. Chem., 1991, vol. 266, pp. 12 289-12 993.
Mironov, A.F., Kozyrev, A.N., and Brandis, A., Sensitizers of Second Generation for Photodynamic Therapy of Cancer Based on Chlorophyll and Bacteriochlorophyll Derivatives, Proc. SPIE-Int. Soc. Opt. Eng., 1992, vol. 1922, pp. 204-208.
Jans, D.A., Ackermann, M., Bischoff, J.R., et al., P34cdc2-Mediated Phosphorylation at T124 Inhibits Nuclear Import of SV40 T-Antigen Proteins, J. Cell Biol., 1991, vol. 115, pp. 1203-1212.
The QIAexpressionist: A Handbook for High-Level Expression and Purification of 6His-Tagged Proteins, 2001, QIAGEN, 5th ed. http://www.qiagen.com/literature/handbooks/qxp/qxp/1016852QXPHB_0301WW.pdf.
Rosenkranz, A.A., Antonenko, Y.N., Smirnova, O.A., et al., Avian Adenovirus Induces Ion Channels in Model Bilayer Lipid Membranes, Biochem. Biophys. Res. Commun., 1997, vol. 236, pp. 750-753.
Hubner, S., Xiao, C.-Y., and Jans, D.A., The Protein Kinase CK2 Site (Ser 111/112) Enhances Recognition of the Simian Virus 40 Large T-Antigen Nuclear Localization Sequence by Importin, J. Biol. Chem., 1997, vol. 272, pp. 17 191-17 195.
Akhlynina, T.V., Rosenkranz, A.A., Jans, D.A., et al., Insulin-Mediated Intracellular Targeting Enhances the Photodynamic Activity of Chlorin e 6, Cancer Res., 1995, vol. 55, pp. 1014-1019.
Finlay, G.J., Baguley, B.C., and Wilson, W.R., A Semiautomated Microculture Method for Investigating Growth Inhibitory Effects of Cytotoxic Compounds on Exponentially Growing Carcinoma Cells, Anal. Biochem., 1984, vol.139, pp. 272-279.
Sahm, U.G., Olivier, G.W., Branch, S.K., et al., Influence of ?-MSH Terminal Amino Acids on Binding Affinity and Biological Activity in Melanoma Cells, Peptides, 1994, vol. 15, pp. 441-446.
Uherek, C., Fominaya, J., and Wels, W., A Modular DNA Carrier Protein Based on the Structure of Diphtheria Toxin Mediates Target Cell-Specific Gene Delivery, J. Biol. Chem., 1998, vol. 273, pp. 8835-8841.
Nir, S. and Nieva, J.L., Interactions of Peptides with Liposomes: Pore Formation and Fusion, Progr. Lipid Res., 2000, vol. 39, pp. 181-206.
Ceresa, B.P. and Schmid, S.L., Regulation of Signal Transduction by Endocytosis, Curr. Opin. Cell Biol., 2000, vol. 12, pp. 204-210.
Chan, C.K. and Jans, D.A., Enhancement of MSHReceptor-and GAL4-Mediated Gene Transfer by Switching the Nuclear Import Pathway, Gene Therapy, 2001, vol.8, pp. 166-171.
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Rosenkranz, A.A., Lunin, V.G., Sergienko, O.V. et al. Targeted Intracellular Site-Specific Drug Delivery: Photosensitizer Targeting to Melanoma Cell Nuclei. Russian Journal of Genetics 39, 198–206 (2003). https://doi.org/10.1023/A:1022488027391
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DOI: https://doi.org/10.1023/A:1022488027391