title: Cellular delivery and biological activity of antisense oligonucleotides conjugated to a targeted protein carrier.
abstract: Targeted delivery can potentially improve the pharmacological effects of antisense and siRNA oligonucleotides. Here, we describe a novel bioconjugation approach to the delivery of splice-shifting antisense oligonucleotides (SSOs). The SSOs are linked to albumin via reversible S-S bonds. The albumin is also conjugated with poly(ethylene glycol) (PEG) chains that terminate in an RGD ligand that selectively binds the alphavbeta3 integrin. As a test system, we utilized human melanoma cells that express the alphavbeta3 integrin and that also contain a luciferase reporter gene that can be induced by delivery of SSOs to the cell nucleus. The RGD-PEG-SSO-albumin conjugates were endocytosed by the cells in an RGD-dependent manner; using confocal fluorescence microscopy, evidence was obtained that the SSOs accumulate in the nucleus. The conjugates were able to robustly induce luciferase expression at concentrations in the 25-200 nM range. At these levels, little short-term or long-term toxicity was observed. Thus, the RGD-PEG-albumin conjugates may provide an effective tool for targeted delivery of oligonucleotides to certain cells and tissues.
authors.affiliation: Division of Molecular Pharmaceutics, School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
authors.forename: Hyunmin
authors.initials: H
authors.lastname: Kang
authors.forename: Md Rowshon
authors.initials: MR
authors.lastname: Alam
authors.forename: Vidula
authors.initials: V
authors.lastname: Dixit
authors.forename: Michael
authors.initials: M
authors.lastname: Fisher
authors.forename: Rudy L
authors.initials: RL
authors.lastname: Juliano
mesh_terms: D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D004705:Endocytosis; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism; D020320:Oligoribonucleotides, Antisense / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology / Q000633:toxicity; D011092:Polyethylene Glycols / Q000737:chemistry; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D009842:Oligopeptides; D020320:Oligoribonucleotides, Antisense; D012709:Serum Albumin; D011092:Polyethylene Glycols; C047981:arginyl-glycyl-aspartic acid; D008156:Luciferases