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.

  • Original Article
  • Published:

Adenovirus-mediated delivery of CD46 attenuates the alternative complement pathway on RPE: implications for age-related macular degeneration

Gene Therapy volume 18, pages 613–621 (2011)Cite this article

Subjects

Abstract

Activation of the alternative pathway of the complement system has been implicated in the pathogenesis of age-related macular degeneration. Membrane attack complex (MAC) has been identified mainly on the Bruch's membrane and drusen underlying the retinal pigment epithelium (RPE). Membrane cofactor protein (CD46) preferentially regulates the alternative pathway of complement. The aim of this study was to evaluate the potential of increasing CD46 expression on RPE cells using an adenovirus as a gene therapy approach to reduce alternative pathway-mediated damage to RPE cells. We generated a recombinant adenovirus vector expressing human CD46 (hCD46) and delivered the vector to murine hepatocytes and RPE cells in vitro. After incubation in human serum in conditions in which the classical pathway of complement was blocked, we measured alternative pathway-mediated damage of these cells by quantifying lysis and MAC formation. Adenovirus expressing hCD46 was delivered to the subretinal space of adult mice, and 1 week later, ocular flat mounts were challenged with human serum and the levels of complement-mediated damage was quantified. Adenovirus-mediated delivery of hCD46 localizes to the basal and lateral surfaces of RPE cells where it offers protection from alternative pathway-mediated damage, but not classical, allowing the classical pathway to function unhindered.

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
Figure 6

Similar content being viewed by others

References

  1. Chevez-Barrios P, Chintagumpala M, Mieler W, Paysse E, Boniuk M, Kozinetz C et al. Response of retinoblastoma with vitreous tumor seeding to adenovirus-mediated delivery of thymidine kinase followed by ganciclovir. J Clin Oncol 2005; 23: 7927–7935.

    Article  CAS  Google Scholar 

  2. Campochiaro PA, Nguyen QD, Shah SM, Klein ML, Holz E, Frank RN et al. Adenoviral vector-delivered pigment epithelium-derived factor for neovascular age-related macular degeneration: results of a phase I clinical trial. Hum Gene Theropy 2006; 17: 167–176.

    Article  CAS  Google Scholar 

  3. Sweigard JH, Cashman SM, Kumar-Singh R . Adenovirus vectors targeting distinct cell types in the retina. Invest Ophthalmol Vis Sci 2010; 51: 2219–2228.

    Article  Google Scholar 

  4. Lamartina S, Cimino M, Roscilli G, Dammassa E, Lazzaro D, Rota R et al. Helper-dependent adenovirus for the gene therapy of proliferative retinopathies: stable gene transfer, regulated gene expression and therapeutic efficacy. J Gene Med 2007; 9: 862–874.

    Article  CAS  Google Scholar 

  5. Kim IH, Józkowicz A, Piedra PA, Oka K, Chan L . Lifetime correction of genetic deficiency in mice with a single injection of helper-dependent adenoviral vector. Proc Natl Acad Sci USA 2001; 98: 13282–13287.

    Article  CAS  Google Scholar 

  6. Parks RJ, Chen L, Anton M, Sankar U, Rudnicki MA, Graham FL . A helper-dependent adenovirus vector system: removal of helper virus by Cre-mediated excision of the viral packaging signal. Proc Natl Acad Sci USA 1996; 93: 13565–13570.

    Article  CAS  Google Scholar 

  7. Kumar-Singh R, Yamashita CK, Tran K, Farber DB . Construction of encapsidated (gutted) adenovirus minichromosomes and their application to rescue of photoreceptor degeneration. Methods Enzymol 2000; 316: 724–743.

    Article  CAS  Google Scholar 

  8. Klein R, Klein BE, Knudtson MD, Meuer SM, Swift M, Gangnon RE . Fifteen-year cumulative incidence of age-related macular degeneration: the Beaver Dam Eye Study. Ophthalmology 2007; 114: 253–262.

    Article  Google Scholar 

  9. van Leeuwen R, Klaver CC, Vingerling JR, Hofman A, de Jong PT . Epidemiology of age-related maculopathy: a review. Eur J Epidemiol 2003; 18: 845–854.

    Article  Google Scholar 

  10. Ozkiris A . Anti-VEGF agents for age-related macular degeneration. Expert Opin Ther Pat 2010; 20: 103–118.

    Article  Google Scholar 

  11. Hageman GS, Luthert PJ, Victor Chong NH, Johnson LV, Anderson DH, Mullins RF . An integrated hypothesis that considers drusen as biomarkers of immune-mediated processes at the RPE-Bruch's membrane interface in aging and age-related macular degeneration. Prog Retin Eye Res 2001; 20: 705–732.

    Article  CAS  Google Scholar 

  12. Johnson LV, Leitner WP, Staples MK, Anderson DH . Complement activation and inflammatory processes in Drusen formation and age related macular degeneration. Exp Eye Res 2001; 73: 887–896.

    Article  CAS  Google Scholar 

  13. Anderson DH, Mullins RF, Hageman GS, Johnson LV . A role for local inflammation in the formation of drusen in the aging eye. Am J Ophthalmol 2002; 134: 411–431.

    Article  CAS  Google Scholar 

  14. Reynolds R, Hartnett ME, Atkinson JP, Giclas PC, Rosner B, Seddon JM . Plasma complement components and activation fragments: associations with age-related macular degeneration genotypes and phenotypes. Invest Ophthalmol Vis Sci 2009; 50: 5818–5827.

    Article  Google Scholar 

  15. Barilla-LaBarca ML, Liszewski MK, Lambris JD, Hourcade D, Atkinson JP . Role of membrane cofactor protein (CD46) in regulation of C4b and C3b deposited on cells. J Immunol 2002; 168: 6298–6304.

    Article  CAS  Google Scholar 

  16. Kavanagh D, Richards A, Atkinson J . Complement regulatory genes and hemolytic uremic syndromes. Annu Rev Med 2008; 59: 293–309.

    Article  CAS  Google Scholar 

  17. Vogt SD, Barnum SR, Curcio CA, Read RW . Distribution of complement anaphylatoxin receptors and membrane-bound regulators in normal human retina. Exp Eye Res 2006; 83: 834–840.

    Article  CAS  Google Scholar 

  18. Ramo K, Cashman SM, Kumar-Singh R . Evaluation of adenovirus-delivered human CD59 as a potential therapy for AMD in a model of human membrane attack complex formation on murine RPE. Invest Ophthalmol Vis Sci 2008; 49: 4126–4136.

    Article  Google Scholar 

  19. Post TW, Liszewski MK, Adams EM, Tedja I, Miller EA, Atkinson JP . Membrane cofactor protein of the complement system: alternative splicing of serine/threonine/proline-rich exons and cytoplasmic tails produces multiple isoforms that correlate with protein phenotype. J Exp Med 1991; 174: 93–102.

    Article  CAS  Google Scholar 

  20. McNearney T, Ballard L, Seya T, Atkinson JP . Membrane cofactor protein of complement is present on human fibroblast, epithelial, and endothelial cells. J Clin Invest 1989; 84: 538–545.

    Article  CAS  Google Scholar 

  21. Edwards AO, Ritter III R, Abel KJ, Manning A, Panhuysen C, Farrer LA . Complement factor H polymorphism and age-related macular degeneration. Science 2005; 308: 421–424.

    Article  CAS  Google Scholar 

  22. Gao LJ, Guo SY, Cai YQ, Gu PQ, Su YJ, Gong H et al. Cooperation of decay-accelerating factor and membrane cofactor protein in regulating survival of human cervical cancer cells. BMC Cancer 2009; 9: 384.

    Article  Google Scholar 

  23. Brodbeck WG, Mold C, Atkinson JP, Medof ME . Cooperation between decay-accelerating factor and membrane cofactor protein in protecting cells from autologous complement attack. J Immunol 2000; 165: 3999–4006.

    Article  CAS  Google Scholar 

  24. Vogt SD, Curcio CA, Wang L, Li CM, McGwin Jr G, Medeiros NE et al. Altered retinal pigment epithelium morphology is associated with decreased expression of complement regulatory protein CD46 and ion transporter MCT3 in geographic atrophy of age-related maculopathy. Invest Ophthalmol Vis Sci 2009; 50: E-Abstract 4180.

  25. Richards A, Kemp EJ, Liszewski MK, Goodship JA, Lampe AK, Decorte R et al. Mutations in human complement regulator, membrane cofactor protein (CD46), predispose to development of familial hemolytic uremic syndrome. Proc Natl Acad Sci USA 2003; 100: 12966–12971.

    Article  CAS  Google Scholar 

  26. Caprioli J, Noris M, Brioschi S, Pianetti G, Castelletti F, Bettinaglio P et al. Genetics of HUS: the impact of MCP, CFH, and IF mutations on clinical presentation, response to treatment, and outcome. Blood 2006; 108: 1267–1279.

    Article  CAS  Google Scholar 

  27. Kavanagh D, Goodship TH . Membrane cofactor protein and factor I: mutations and transplantation. Semin Thromb Hemost 2006; 32: 155–159.

    Article  CAS  Google Scholar 

  28. Fallaux FJ, Kranenburg O, Cramer SJ, Houweling A, Van Ormondt H, Hoeben RC et al. Characterization of 911: a new helper cell line for the titration and propagation of early region 1-deleted adenoviral vectors. Hum Gene Theropy 1996; 7: 215–222.

    Article  CAS  Google Scholar 

  29. He TC, Zhou S, da Costa LT, Yu J, Kinzler KW, Vogelstein B . A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci USA 1998; 95: 2509–2514.

    Article  CAS  Google Scholar 

  30. Cashman SM, Morris DJ, Kumar-Singh R . Adenovirus type 5 pseudotyped with adenovirus type 37 fiber uses sialic acid as a cellular receptor. Virology 2004; 324: 129–139.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by grants to RK-S from The Ellison Foundation, The Virginia B Smith Trust and grants to the Department of Ophthalmology at Tufts University from the Lions Eye Foundation and Research to Prevent Blindness.

Author information

Authors and Affiliations

  1. Department of Ophthalmology and Neuroscience, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, MA, USA

    J H Sweigard, S M Cashman & R Kumar-Singh

  2. Program in Cell, Molecular and Developmental Biology, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, MA, USA

    J H Sweigard & R Kumar-Singh

Authors
  1. J H Sweigard
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S M Cashman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R Kumar-Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to R Kumar-Singh.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sweigard, J., Cashman, S. & Kumar-Singh, R. Adenovirus-mediated delivery of CD46 attenuates the alternative complement pathway on RPE: implications for age-related macular degeneration. Gene Ther 18, 613–621 (2011). https://doi.org/10.1038/gt.2011.6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/gt.2011.6

Keywords

Search

Advanced search

Quick links