Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Stable rAAV-mediated transduction of rod and cone photoreceptors in the canine retina

Gene Therapy volume 10, pages 1336–1344 (2003)Cite this article

Abstract

Recombinant adeno-associated virus (rAAV) vectors are attractive candidates for the treatment of inherited and acquired retinal disease. Although rAAV vectors are well characterized in rodent models, a prerequisite to their clinical application in human patients is the thorough evaluation of their efficacy and safety in intermediate animal models. In this study, we describe rAAV-2-mediated expression of GFP reporter gene in retinal cells following local vector delivery in dogs. Subretinal delivery of rAAV.CMV.GFP was performed unilaterally in eight normal dogs from 6 weeks of age. The area of retinal transduction was maximized by the optimization of surgical techniques for subretinal vector delivery by pars-plana vitrectomy and the use of fine-gauge subretinal cannulae to create multiple retinotomies. rAAV-2 vectors mediated efficient stable reporter gene expression in photoreceptors and retinal pigment epithelial cells. We found efficient transduction of cone photoreceptors in addition to rods in both the canine retina and after subretinal vector delivery in another intermediate animal model, the feline retina. GFP expression in dogs was confined to the area of the retinal bleb and was sustained in cells at this site for at least 18 months. Electroretinography demonstrated a modest reduction in global rod-mediated retinal function following subretinal delivery of rAAV.CMV.GFP. Three of the eight animals developed delayed-onset intraocular inflammation, in two cases associated with a serum antibody response to GFP protein. We conclude that rAAV-2 vectors mediate efficient sustained transgene expression in rod and cone photoreceptors following subretinal delivery in this intermediate animal model. The possibility of adverse effects including intraocular immune responses and reduced retinal function requires further investigation prior to clinical applications in patients.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Bessant DA, Ali RR, Bhattacharya SS . Molecular genetics and prospects for therapy of the inherited retinal dystrophies. Curr Opin Genet Dev 2001; 11: 307–316.

    Article  CAS  PubMed  Google Scholar 

  2. Rahmani B et al. The cause-specific prevalence of visual impairment in an urban population. The Baltimore Eye Survey. Ophthalmology 1996; 103: 1721–1726.

    Article  CAS  PubMed  Google Scholar 

  3. Bressler NM, Bressler SB . Preventative ophthalmology. Age-related macular degeneration. Ophthalmology 1995; 102: 1206–1211.

    Article  CAS  PubMed  Google Scholar 

  4. Early Treatment Diabetic Retinopathy Study Research Group. Early photocoagulation for diabetic retinopathy. ETDRS report number 9. Ophthalmology 1991; 98: 766–785.

  5. Macular Photocoagulation Study Group. Laser photocoagulation of subfoveal neovascular lesions in age-related macular degeneration. Results of a randomized clinical trial. [see comments]. Arch Ophthalmol 1991; 109: 1220–1231.

  6. Ali RR et al. Gene transfer into the mouse retina mediated by an adeno-associated viral vector. Hum Mol Genet 1996; 5: 591–594.

    Article  CAS  PubMed  Google Scholar 

  7. Bennett J, Duan D, Engelhardt JF, Maguire AM . Real-time, noninvasive in vivo assessment of adeno-associated virus-mediated retinal transduction. Invest Ophthalmol Vis Sci 1997; 38: 2857–2863.

    CAS  PubMed  Google Scholar 

  8. Ali RR et al. Adeno-associated virus gene transfer to mouse retina. Hum Gene Therapy 1998; 9: 81–86.

    Article  CAS  Google Scholar 

  9. Sarra GM et al. Gene replacement therapy in the retinal degeneration slow (rds) mouse: the effect on retinal degeneration following partial transduction of the retina. Hum Mol Genet 2001; 10: 2353–2361.

    Article  CAS  PubMed  Google Scholar 

  10. Bennett J et al. Stable transgene expression in rod photoreceptors after recombinant adeno-associated virus-mediated gene transfer to monkey retina. Proc Natl Acad Sci USA 1999; 96: 9920–9925.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Ali RR et al. Restoration of photoreceptor ultrastructure and function in retinal degeneration slow mice by gene therapy. Nat Genet 2000; 25: 306–310.

    Article  CAS  PubMed  Google Scholar 

  12. Bainbridge JW et al. Inhibition of retinal neovascularisation by gene transfer of soluble VEGF receptor sFlt-1. Gene Therapy 2002; 9: 320–326.

    Article  CAS  PubMed  Google Scholar 

  13. Mori K et al. AAV-mediated gene transfer of pigment epithelium-derived factor inhibits choroidal neovascularization. Invest Ophthalmol Vis Sci 2002; 43: 1994–2000.

    PubMed  Google Scholar 

  14. Lai YK et al. Potential long-term inhibition of ocular neovascularisation by recombinant adeno-associated virus-mediated secretion gene therapy. Gene Ther 2002; 9: 804–813.

    Article  CAS  PubMed  Google Scholar 

  15. Dudus L et al. Persistent transgene product in retina, optic nerve and brain after intraocular injection of rAAV. Vision Res 1999; 39: 2545–2553.

    Article  CAS  PubMed  Google Scholar 

  16. Acland GM et al. Gene therapy restores vision in a canine model of childhood blindness. Nat Genet 2001; 28: 92–95.

    CAS  PubMed  Google Scholar 

  17. Bennett J, Anand V, Acland GM, Maguire AM . Cross-species comparison of in vivo reporter gene expression after recombinant adeno-associated virus-mediated retinal transduction. Methods Enzymol 2000; 316: 777–789.

    Article  CAS  PubMed  Google Scholar 

  18. Zhang X et al. High-titer recombinant adeno-associated virus production from replicating amplicons and herpes vectors deleted for glycoprotein H. Hum Gene Ther 1999; 10: 2527–2537.

    Article  CAS  PubMed  Google Scholar 

  19. Conway JE et al. High-titer recombinant adeno-associated virus production utilizing a recombinant herpes simplex virus type I vector expressing AAV-2 Rep and Cap. Gene Therapy 1999; 6: 986–993.

    Article  CAS  PubMed  Google Scholar 

  20. Blanks JC, Johnson LV . Specific binding of peanut lectin to a class of retinal photoreceptor cells. A species comparison. Invest Ophthalmol Vis Sci 1984; 25: 546–557.

    CAS  PubMed  Google Scholar 

  21. Greiner JV, Weidman TA . Histogenesis of the ferret retina. Exp Eye Res 1981; 33: 315–332.

    Article  CAS  PubMed  Google Scholar 

  22. Lotery AJ et al. Gene transfer to the nonhuman primate retina with recombinant feline immunodeficiency virus vectors. Hum Gene Ther 2002; 13: 689–696.

    Article  CAS  PubMed  Google Scholar 

  23. Imai K, Loewenstein A, de Juan Jr E . Translocation of the retina for management of subfoveal choroidal neovascularization I: experimental studies in the rabbit eye. Am J Ophthalmol 1998; 125: 627–634.

    Article  CAS  PubMed  Google Scholar 

  24. Terasaki H et al. Change in full-field ERGs after macular translocation surgery with 360 degrees retinotomy. Invest Ophthalmol Vis Sci 2002; 43: 452–457.

    PubMed  Google Scholar 

  25. Luke C et al. Electrophysiological changes after 360 degrees retinotomy and macular translocation for subfoveal choroidal neovascularisation in age related macular degeneration. Br J Ophthalmol 2001; 85: 928–932.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We gratefully acknowledge the advice and assistance of Paul Sullivan, David Williams, David Gould, Rachael Bennett and Phil Luthert. JWBB is a Wellcome Trust Research Training Fellow. This work was supported by the British Retinitis Pigmentosa Society, the Sir Jules Thorn Charitable Trust and Fight for Sight.

Author information

Authors and Affiliations

  1. Department of Molecular Genetics, Institute of Ophthalmology, University College London, London, UK

    J W B Bainbridge, F C Schlichtenbrede, A Smith, C Broderick & R R Ali

  2. Institute of Child Health, University College London, Guilford Street, London, UK

    A Mistry, M De Alwis, A Georgiadis, A J Thrasher & R R Ali

  3. Department of Clinical Veterinary Medicine, University of Cambridge, Madlingley Road, Cambridge, UK

    P M Taylor, M Squires & D Sargan

  4. Department of Pathology, Institute of Ophthalmology, University College London, London, UK

    C Sethi & D Charteris

Authors
  1. J W B Bainbridge
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A Mistry
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. F C Schlichtenbrede
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C Broderick
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M De Alwis
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A Georgiadis
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. P M Taylor
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. M Squires
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. C Sethi
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. D Charteris
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. A J Thrasher
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. D Sargan
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. R R Ali
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bainbridge, J., Mistry, A., Schlichtenbrede, F. et al. Stable rAAV-mediated transduction of rod and cone photoreceptors in the canine retina. Gene Ther 10, 1336–1344 (2003). https://doi.org/10.1038/sj.gt.3301990

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301990

Keywords

This article is cited by

Search

Advanced search

Quick links