Sequence dependence of C5-propynyl-dU,dC-phosphorothioate oligonucleotide inhibition of the human IGF-I receptor: mRNA, protein, and cell growth

Rhys Fogarty, Sandra C. McKean, Paul J. White, Lynne M. Atley, George A. Werther, Christopher J. Wraight

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Human keratinocytes are highly responsive to mitogenic and antiapoptotic signaling by the insulinlike growth factor-I receptor (IGF-IR). IGF-IR hyperstimulation is a feature of hyperplastic skin conditions, making the IGF-IR an appealing target for antisense therapeutic intervention. In this study, we used a C5-propynyl-dU,dC-phosphorothioate oligo-2′-deoxyribonucleotide antisense 15-mer to the human IGF-IR mRNA, along with liposome transfection, to inhibit IGF-IR activity in a human keratinocyte cell line and demonstrated potent inhibition of cell growth despite the presence of serum. To investigate the sequence specificity of these effects and to establish the concentration range over which a purely antisense effect could be demonstrated, we introduced 1, 2, 4, 8, and 15 base mismatches into the oligonucleotide and analyzed changes in inhibitory efficacy. In the 10-30 nM concentration range, the introduction of 1 and 2 mismatches into the middle of the 15-mer only modestly affected inhibitory efficacy, whereas >4 mismatches profoundly reduced mRNA, protein, and growth-inhibitory effects. From these results, we conclude that (1) sequence-specific antisense inhibition of IGF-IR activity in keratinocytes is achievable, (2) potent anti-IGF-IR antisense inhibition can be achieved in vitro at concentrations as low as 10 nM, and (3) a sequence-dependent mechanism is likely to underpin the observed in vivo therapeutic effects (Wraight et al. Nat. Biotechnol. 2000;18:521) of these antisense oligonucleotides (AS-ODN) in cutaneous hyperplastic disorders, such as psoriasis.

LanguageEnglish
Pages369-377
Number of pages9
JournalAntisense and Nucleic Acid Drug Development
Volume12
Issue number6
DOIs
Publication statusPublished - 1 Jan 2002
Externally publishedYes

ASJC Scopus subject areas

  • Genetics
  • Pharmacology

Cite this

Fogarty, Rhys ; McKean, Sandra C. ; White, Paul J. ; Atley, Lynne M. ; Werther, George A. ; Wraight, Christopher J. / Sequence dependence of C5-propynyl-dU,dC-phosphorothioate oligonucleotide inhibition of the human IGF-I receptor : mRNA, protein, and cell growth. In: Antisense and Nucleic Acid Drug Development. 2002 ; Vol. 12, No. 6. pp. 369-377.
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Sequence dependence of C5-propynyl-dU,dC-phosphorothioate oligonucleotide inhibition of the human IGF-I receptor : mRNA, protein, and cell growth. / Fogarty, Rhys; McKean, Sandra C.; White, Paul J.; Atley, Lynne M.; Werther, George A.; Wraight, Christopher J.

In: Antisense and Nucleic Acid Drug Development, Vol. 12, No. 6, 01.01.2002, p. 369-377.

Research output: Contribution to journalArticle

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T1 - Sequence dependence of C5-propynyl-dU,dC-phosphorothioate oligonucleotide inhibition of the human IGF-I receptor

T2 - Antisense and Nucleic Acid Drug Development

AU - Fogarty, Rhys

AU - McKean, Sandra C.

AU - White, Paul J.

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AU - Wraight, Christopher J.

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