title: Recombinant modular transporters for cell-specific nuclear delivery of locally acting drugs enhance photosensitizer activity.
abstract: The search for new pharmaceuticals that are specific for diseased rather than normal cells in the case of cancer and viral disease has raised interest in locally acting drugs that act over short distances within the cell and for which different cell compartments have distinct sensitivities. Thus, photosensitizers (PSs) used in anti-cancer therapy should ideally be transported to the most sensitive subcellular compartments in order for their action to be most pronounced. Here we describe the design, production, and characterization of the effects of bacterially expressed modular recombinant transporters for PSs comprising 1) alpha-melanocyte-stimulating hormone as an internalizable, cell-specific ligand; 2) an optimized nuclear localization sequence of the SV40 large T-antigen; 3) an Escherichia coli hemoglobin-like protein as a carrier; and 4) an endosomolytic amphipathic polypeptide, the translocation domain of diphtheria toxin. These modular transporters delivered PSs into the nuclei, the most vulnerable sites for the action of PSs, of murine melanoma cells, but not non-MSH receptor-overexpressing cells, to result in cytotoxic effects several orders of magnitude greater than those of nonmodified PSs. The modular fusion proteins described here for the first time, capable of cell-specific targeting to particular subcellular compartments to increase drug efficacy, represent new pharmaceuticals with general application.
authors.affiliation: Department of Molecular Genetics of Intracellular Transport, Institute of Gene Biology, Russian Academy of Sciences, 34/5 Vavilov St., 119334, Moscow, Russia.
authors.forename: Andrey A
authors.initials: AA
authors.lastname: Rosenkranz
authors.forename: Vladimir G
authors.initials: VG
authors.lastname: Lunin
authors.forename: Pavel V
authors.initials: PV
authors.lastname: Gulak
authors.forename: Olga V
authors.initials: OV
authors.lastname: Sergienko
authors.forename: Maria A
authors.initials: MA
authors.lastname: Shumiantseva
authors.forename: Olga L
authors.initials: OL
authors.lastname: Voronina
authors.forename: Dinara G
authors.initials: DG
authors.lastname: Gilyazova
authors.forename: Anna P
authors.initials: AP
authors.lastname: John
authors.forename: Anna A
authors.initials: AA
authors.lastname: Kofner
authors.forename: Andrey F
authors.initials: AF
authors.lastname: Mironov
authors.forename: David A
authors.initials: DA
authors.lastname: Jans
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Sobolev
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000378:metabolism / Q000627:therapeutic use; D001692:Biological Transport; D002467:Cell Nucleus / Q000187:drug effects* / Q000378:metabolism; D004093:Dihydropteridine Reductase / Q000235:genetics; D004337:Drug Carriers / Q000378:metabolism; D016503:Drug Delivery Systems / Q000379:methods*; D029968:Escherichia coli Proteins / Q000235:genetics; D006420:Hemeproteins / Q000235:genetics; D008546:Melanoma, Experimental / Q000188:drug therapy / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics; D019913:Nuclear Localization Signals; D010778:Photochemotherapy / Q000379:methods*; D017319:Photosensitizing Agents / Q000008:administration & dosage* / Q000378:metabolism / Q000627:therapeutic use; D010957:Plasmids; D011166:Porphyrins / Q000008:administration & dosage / Q000378:metabolism / Q000627:therapeutic use; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000378:metabolism / Q000627:therapeutic use; D014407:Tumor Cells, Cultured; D000521:alpha-MSH / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D004337:Drug Carriers; D029968:Escherichia coli Proteins; D006420:Hemeproteins; D019913:Nuclear Localization Signals; D017319:Photosensitizing Agents; D011166:Porphyrins; D011993:Recombinant Fusion Proteins; C452433:bacteriochlorin; D000521:alpha-MSH; D004093:Dihydropteridine Reductase; C076538:hmp protein, E coli; D009247:NADH, NADPH Oxidoreductases