Targeted delivery of antisense oligonucleotides by chemically self-assembled nanostructures

Amit Gangar, Adrian Fegan, Sidath C Kumarapperuma, Peter Huynh, Alexey Benyumov, Carston R Wagner

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Synthetic nucleic acids have shown great potential in the treatment of various diseases. Nevertheless, the selective delivery to a target tissue has proved challenging. The coupling of nucleic acids to targeting peptides, proteins, and antibodies has been explored as an approach for their selective tissue delivery. Nevertheless, the preparation of covalently coupled peptides and proteins that can also undergo intracellular release as well as deliver more than one copy of the nucleic acid has proved challenging. Recently, we have developed a novel method for the rapid noncovalent conjugation of nucleic acids to targeting single chain antibodies (scFv) using chemically self-assembled nanostructures (CSANs). CSANs have been prepared by the self-assembly of two dihydrofolate reductase molecules (DHFR2) and a targeting scFv in the presence of bis-methotrexate (bis-MTX). The valency of the nanorings can be tuned from one to eight subunits, depending on the length and composition of the linker between the dihydrofolate reductase molecules. To explore their potential for the therapeutic delivery of nucleic acids as well as the ability to expand the capabilities of CSANs by incorporating smaller cyclic targeting peptides, we prepared DHFR2 proteins fused through a flexible peptide linker to cyclic-RGD, which targets αvβ3 integrins, and a bis-MTX chemical dimerizer linked to an antisense oligonucleotide (bis-MTX-ASO) that has been shown to silence expression of eukaryotic translation initiation factor 4E (eIF4E). Monomeric and multimeric cRGD-CSANs were then prepared with bis-MTX-ASO and shown to undergo endocytosis in the breast cancer cell line, MDA-MB-231, which overexpresses αvβ3. The bis-MTX-ASO was shown to undergo endosomal escape resulting in the knock down of eIF4E with at least the same efficiency as ASO delivered by oligofectamine. The modularity, flexibility, and common method of conjugation may prove to be a useful general approach for the targeted delivery of ASOs, as well as other nucleic acids to cells.

Original languageEnglish (US)
Pages (from-to)3514-3518
Number of pages5
JournalMolecular Pharmaceutics
Volume10
Issue number9
DOIs
StatePublished - Sep 3 2013

Fingerprint

Nanostructures
Antisense Oligonucleotides
Nucleic Acids
Methotrexate
Eukaryotic Initiation Factor-4E
Eukaryotic Initiation Factors
Tetrahydrofolate Dehydrogenase
Peptides
Single-Chain Antibodies
Cyclic Peptides
Protein Transport
Endocytosis
Integrins
Proteins
Breast Neoplasms
Cell Line
Antibodies

Keywords

  • Antisense
  • Delivery
  • Nanotechnology
  • Nucleic acid

Cite this

Targeted delivery of antisense oligonucleotides by chemically self-assembled nanostructures. / Gangar, Amit; Fegan, Adrian; Kumarapperuma, Sidath C; Huynh, Peter; Benyumov, Alexey; Wagner, Carston R.

In: Molecular Pharmaceutics, Vol. 10, No. 9, 03.09.2013, p. 3514-3518.

Research output: Contribution to journalArticle

Gangar, Amit ; Fegan, Adrian ; Kumarapperuma, Sidath C ; Huynh, Peter ; Benyumov, Alexey ; Wagner, Carston R. / Targeted delivery of antisense oligonucleotides by chemically self-assembled nanostructures. In: Molecular Pharmaceutics. 2013 ; Vol. 10, No. 9. pp. 3514-3518.
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