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Reduction of fibronectin expression by intravitreal administration of antisense oligonucleotides

Nature Biotechnology volume 17, pages 476–479 (1999)Cite this article

Abstract

We have investigated whether antisense oligonucleotides delivered intravitreally could reduce gene expression specifically in the retina. In this study, phosphorothioate antisense oligonucleotides targeted to fibronectin transcripts were coupled to a novel carrier and used to specifically reduce fibronectin (FN) expression in retinal vascular cells. Using confocal microscopy, fluorescence from fluorescein isothiocyanate-labeled FN-oligonucleotides was detected in retinal vascular cells at 24 h postinjection and persisted until day 6 (the end point of this study). The fibronectin mRNA level was consistently decreased to 86.7% ± 7.9% of control (p<0.05) at day 2, and 46.7% ± 4.9% of control (p<0.01) at day 6. In contrast, the β-actin mRNA level, an internal control, was unaltered in rat retinas that received FN-oligonucleotides. Fibronectin protein level at day 6 was also significantly reduced to 61.4% ± 16% of control (p<0.01). No toxic effect resulting from the carrier was detected histologically. Thus, intravitreal delivery of antisense oligonucleotides to modulate abnormal gene expression in retinal diseases may be an effective approach for ocular gene therapy.

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Figure 1: Distribution of antisense FN-oligonucleotides in rat retina.
Figure 2: Localization of antisense FN-oligonucleotides in rat retinal vascular cells.
Figure 3: Relative levels of fibronectin mRNA and protein in rat retinas.

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Acknowledgements

This work was supported by a Research Award from the American Diabetes Association and the National Eye Institute, NIH (EY11990-01A1). The work of the group at the University of Nebraska Medical Center was supported by a grant from the National Science Foundation (BES-9712657).

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Authors and Affiliations

  1. Schepens Eye Research Institute, Harvard Medical School , Boston, 02114, MA

    Sayon Roy, Kathy Zhang & Richard S. Kao

  2. University of Massachusetts Medical School, Worcester, 01655, MA

    Timothy Roth

  3. University of Nebraska Medical Center, Omaha, 68198, NE

    Sergei Vinogradov & Alexander Kabanov

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  1. Sayon Roy
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Correspondence to Sayon Roy.

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Roy, S., Zhang, K., Roth, T. et al. Reduction of fibronectin expression by intravitreal administration of antisense oligonucleotides. Nat Biotechnol 17, 476–479 (1999). https://doi.org/10.1038/8654

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