Cellular delivery and biological activity of antisense oligonucleotides conjugated to a targeted protein carrier.
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.
Kang
Hyunmin
H
Division of Molecular Pharmaceutics, School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
Alam
Md Rowshon
MR
Dixit
Vidula
V
Fisher
Michael
M
Juliano
Rudy L
RL
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*
D009842:Oligopeptides; D020320:Oligoribonucleotides, Antisense; D012709:Serum Albumin; D011092:Polyethylene Glycols; C047981:arginyl-glycyl-aspartic acid; D008156:Luciferases