Short Communication | Published:

In situ regeneration of retinal pigment epithelium by gene transfer of E2F2: a potential strategy for treatment of macular degenerations

Gene Therapy volume 24, pages 810818 (2017) | Download Citation

Abstract

The retinal pigment epithelium (RPE) interacts closely with photoreceptors to maintain visual function. In degenerative diseases such as Stargardt disease and age-related macular degeneration, the leading cause of blindness in the developed world, RPE cell loss is followed by photoreceptor cell death. RPE cells can proliferate under certain conditions, suggesting an intrinsic regenerative potential, but so far this has not been utilised therapeutically. Here, we used E2F2 to induce RPE cell replication and thereby regeneration. In both young and old (2 and 18 month) wildtype mice, subretinal injection of non-integrating lentiviral vector expressing E2F2 resulted in 47% of examined RPE cells becoming BrdU positive. E2F2 induced an increase in RPE cell density of 17% compared with control vector-treated and 14% compared with untreated eyes. We also tested this approach in an inducible transgenic mouse model of RPE loss, generated through activation of diphtheria toxin-A gene. E2F2 expression resulted in a 10-fold increase in BrdU uptake and a 34% increase in central RPE cell density. Although in mice this localised rescue is insufficiently large to be demonstrable by electroretinography, a measure of massed retinal function, these results provide proof-of-concept for a strategy to induce in situ regeneration of RPE for the treatment of RPE degeneration.

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References

  1. 1.

    , , , . Age-related macular degeneration–emerging pathogenetic and therapeutic concepts. Ann Med 2006; 38: 450–471.

  2. 2.

    , , , . The five-year incidence and progression of age-related maculopathy: the Beaver Dam Eye Study. Ophthalmology 1997; 104: 7–21.

  3. 3.

    , , . Prevalence of age-related maculopathy. The Beaver Dam Eye Study. Ophthalmology 1992; 99: 933–943.

  4. 4.

    , , , , , et al. Prevalence of age-related macular degeneration in the United States. Arch Ophthalmol 2004; 122: 564–572.

  5. 5.

    . The retinal pigment epithelium in visual function. Physiol Rev 2005; 85: 845–881.

  6. 6.

    , , , , . Age-related macular degeneration: etiology, pathogenesis, and therapeutic strategies. Surv Ophthalmol 2003; 48: 257–293.

  7. 7.

    , , , , . Cell loss in the aging retina. Relationship to lipofuscin accumulation and macular degeneration. Invest Ophthalmol Vis Sci 1989; 30: 1691–1699.

  8. 8.

    . The ageing retina: physiology or pathology. Eye (Lond) 1987; 1 (Pt 2): 282–295.

  9. 9.

    . [Mitotic activity of the pigment epithelium during embryonic and postembryonic development]. Adv Ophthalmol 1979; 39: 37–58.

  10. 10.

    , , . Age-related changes in human RPE cell density and apoptosis proportion in situ. Invest Ophthalmol Vis Sci 2002; 43: 3312–3318.

  11. 11.

    , , , , , . Genetic ablation of retinal pigment epithelial cells reveals the adaptive response of the epithelium and impact on photoreceptors. Proc Natl Acad Sci USA 2009; 106: 18728–18733.

  12. 12.

    , . Retinal separation, retinotomy, and macular relocation: II. A surgical approach for age-related macular degeneration? Graefes Arch Clin Exp Ophthalmol 1993; 231: 635–641.

  13. 13.

    , , . Macular translocation for neovascular age-related macular degeneration. Cochrane Database Syst Rev 2008; 8: CD006928.

  14. 14.

    , , , . Long-term results of submacular surgery combined with macular translocation of the retinal pigment epithelium in neovascular age-related macular degeneration. Ophthalmology 2005; 112: 2081–2087.

  15. 15.

    , , , , . RPE transplantation and its role in retinal disease. Prog Retin Eye Res 2007; 26: 598–635.

  16. 16.

    , , , . Transplantation of the RPE in AMD. Prog Retin Eye Res 2007; 26: 516–554.

  17. 17.

    , , , , . Long-term results of full macular translocation for choroidal neovascularization in age-related macular degeneration. Ophthalmology 2015; 122: 1366–1374.

  18. 18.

    , , , , , et al. Iris pigment epithelial translocation in the treatment of exudative macular degeneration: a 3-year follow-up. Arch Ophthalmol 2006; 124: 183–188.

  19. 19.

    , , , , , et al. Autologous translocation of the choroid and retinal pigment epithelium in patients with geographic atrophy. Ophthalmology 2007; 114: 551–560.

  20. 20.

    , , , , , et al. Autologous transplantation of the retinal pigment epithelium and choroid in the treatment of neovascular age-related macular degeneration. Ophthalmology 2007; 114: 561–570.

  21. 21.

    , , , , . Comparison of the growth potential of retinal pigment epithelial cells obtained during vitrectomy in patients with age-related macular degeneration or complex retinal detachment. Graefes Arch Clin Exp Ophthalmol 2004; 242: 442–443.

  22. 22.

    , , , , . Transplantation of cultured human retinal epithelium to Bruch's membrane of the owl monkey's eye. Curr Eye Res 1985; 4: 253–265.

  23. 23.

    , , , , , et al. Human embryonic stem cell-derived retinal pigment epithelium in patients with age-related macular degeneration and Stargardt's macular dystrophy: follow-up of two open-label phase 1/2 studies. Lancet 2015; 385: 509–516.

  24. 24.

    . Stem cells cruise to clinic. Nature 2013; 494: 413.

  25. 25.

    , , , . Perspectives on human clinical trials of therapies using iPS cells in Japan: reaching the forefront of stem-cell therapies. Biosci Trends 2013; 7: 157–158.

  26. 26.

    . The regulation of E2F by pRB-family proteins. Genes Dev 1998; 12: 2245–2262.

  27. 27.

    . The Rb/E2F pathway and cancer. Hum Mol Genet 2001; 10: 699–703.

  28. 28.

    , , , , , et al. The E2F1-3 transcription factors are essential for cellular proliferation. Nature 2001; 414: 457–462.

  29. 29.

    , , , , . Distinct roles for E2F proteins in cell growth control and apoptosis. Proc Natl Acad Sci USA 1997; 94: 7245–7250.

  30. 30.

    , , . In vitro growth of pure cultures of retinal pigment epithelium. Arch Ophthalmol 1972; 88: 63–69.

  31. 31.

    , . Pigment epithelium proliferation in retinal detachment (massive periretinal proliferation). Am J Ophthalmol 1975; 80: 1–23.

  32. 32.

    , , . Pigment epithelial proliferation in human retinal detachment with massive periretinal proliferation. Am J Ophthalmol 1978; 85: 181–191.

  33. 33.

    , , , , . The onset of pigment epithelial proliferation after retinal detachment. Invest Ophthalmol Vis Sci 1981; 21: 10–16.

  34. 34.

    , , , , , et al. Quantitative autofluorescence and cell density maps of the human retinal pigment epithelium. Invest Ophthalmol Vis Sci 2014; 55: 4832–4841.

  35. 35.

    , , , , . Mature retinal pigment epithelium cells are retained in the cell cycle and proliferate in vivo. Mol Vis 2008; 14: 1784–1791.

  36. 36.

    , , , , , et al. The tight junction associated signalling proteins ZO-1 and ZONAB regulate retinal pigment epithelium homeostasis in mice. PLoS One 2010; 5: e15730.

  37. 37.

    , , . Epithelial-mesenchymal transition and proliferation of retinal pigment epithelial cells initiated upon loss of cell-cell contact. Invest Ophthalmol Vis Sci 2010; 51: 2755–2763.

  38. 38.

    , , , . E2F target genes: unraveling the biology. Trends Biochem Sci 2004; 29: 409–417.

  39. 39.

    , , , , , et al. The cyclin D1-CDK4 oncogenic interactome enables identification of potential novel oncogenes and clinical prognosis. Cell Cycle 2014; 13: 2889–2900.

  40. 40.

    , , , . Cellular restriction of retrovirus particle-mediated mRNA transfer. J Virol 2008; 82: 3069–3077.

  41. 41.

    , , , , , et al. High-level transduction and gene expression in hematopoietic repopulating cells using a human immunodeficiency [correction of imunodeficiency] virus type 1-based lentiviral vector containing an internal spleen focus forming virus promoter. Hum Gene Ther 2002; 13: 803–813.

  42. 42.

    , , , , , et al. Effective gene therapy with nonintegrating lentiviral vectors. Nat Med 2006; 12: 348–353.

  43. 43.

    , , , , , et al. In vivo gene transfer to the mouse eye using an HIV-based lentiviral vector; efficient long-term transduction of corneal endothelium and retinal pigment epithelium. Gene Ther 2001; 8: 1665–1668.

  44. 44.

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

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Acknowledgements

This work was supported by the National Institute for Health Research Biomedical Research Centre at Moorfields Eye Hospital and UCL, and also by Moorfields Eye Charity and RP Fighting Blindness. The authors gratefully acknowledge Anselm Kampik, Augenzentrum im Brienner Hof, Munich, Germany, for helpful discussions. The authors thank Nancy Joyce, Schepens Eye Research Institute, Harvard Medical School, Boston, USA, for providing the human E2F2 cDNA plasmid.

Author information

Affiliations

  1. Department of Genetics, UCL Institute of Ophthalmology, London, UK

    • D Kampik
    • , M Basche
    • , U F O Luhmann
    • , K M Nishiguchi
    • , S Azam
    • , Y Duran
    • , S J Robbie
    • , J W B Bainbridge
    • , A J Smith
    •  & R R Ali
  2. University Hospital of Würzburg, Department of Ophthalmology, Würzburg, Germany

    • D Kampik
    •  & H Han
  3. Roche Pharmaceutical Research and Early Development, Ophthalmology Discovery & Biomarkers, Basel, Switzerland

    • U F O Luhmann
  4. Current address: Department of Advanced Ophthalmic Medicine, Graduate School of Medicine, Tohoku University, Sendai, Japan

    • K M Nishiguchi
  5. Department of Cell Biology, UCL Institute of Ophthalmology, London, UK

    • J A E Williams
    • , J Greenwood
    •  & S E Moss
  6. Moorfields Eye Hospital, London, UK

    • J W B Bainbridge
    •  & D F Larkin
  7. NIHR Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, UK

    • R R Ali

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Competing interests

UFOL is employee of F Hoffmann-La Roche Ltd. The remaning authors declare no conflict interest.

Corresponding author

Correspondence to R R Ali.

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DOI

https://doi.org/10.1038/gt.2017.89

Supplementary Information accompanies this paper on Gene Therapy website (http://www.nature.com/gt)