Cellular Delivery and Biological Activity of Antisense Oligonucleotides Conjugated to a Targeted Protein Carrier

Hyunmin Kang, Md. Rowshon Alam, Vidula Dixit, Michael Fisher and Rudy L. Juliano*
Division of Molecular Pharmaceutics, School of Pharmacy, CB 7360, University of North Carolina, Chapel Hill, North Carolina 27599
Bioconjugate Chem., 2008, 19 (11), pp 2182–2188
DOI: 10.1021/bc800270w
Publication Date (Web): October 1, 2008
Copyright © 2008 American Chemical Society
* Corresponding author. arjay@med.unc.edu, phone, 919 966 4383.
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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 αvβ3 integrin. As a test system, we utilized human melanoma cells that express the αvβ3 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.

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History

  • Published In Issue November 19, 2008
  • Article ASAPOctober 01, 2008
  • Received: July 1, 2008
    Revised: September 5, 2008

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