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An animal model of age-related macular degeneration in senescent Ccl-2- or Ccr-2-deficient mice

Nature Medicine volume 9, pages 1390–1397 (2003)Cite this article

Abstract

The study and treatment of age-related macular degeneration (AMD), a leading cause of blindness, has been hampered by a lack of animal models. Here we report that mice deficient either in monocyte chemoattractant protein-1 (Ccl-2; also known as MCP-1) or its cognate C-C chemokine receptor-2 (Ccr-2) develop cardinal features of AMD, including accumulation of lipofuscin in and drusen beneath the retinal pigmented epithelium (RPE), photoreceptor atrophy and choroidal neovascularization (CNV). Complement and IgG deposition in RPE and choroid accompanies senescence in this model, as in human AMD. RPE or choroidal endothelial production of Ccl-2 induced by complement C5a and IgG may mediate choroidal macrophage infiltration into aged wild-type choroids. Wild-type choroidal macrophages degrade C5 and IgG in eye sections of Ccl2−/− or Ccr2−/− mice. Impaired macrophage recruitment may allow accumulation of C5a and IgG, which induces vascular endothelial growth factor (VEGF) production by RPE, possibly mediating development of CNV. These models implicate macrophage dysfunction in AMD pathogenesis and may be useful as a platform for validating therapies.

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Figure 1: Ccl2−/− and Ccr2−/− mice develop early AMD.
Figure 2: Ccl2−/− and Ccr2−/− mice develop retinal degeneration.
Figure 3: Ccl2−/− and Ccr2−/− mice develop neovascular AMD and overexpress VEGF in RPE.
Figure 4: Complement and IgG deposition can stimulate Ccl-2 and VEGF secretion.
Figure 5: Ccl-2 overexpression and macrophage infiltration in aged wild-type mice.
Figure 6: Macrophages are immobilized by, adhere to and degrade C5 and IgG.

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References

  1. Smith, W. et al. Risk factors for age-related macular degeneration: pooled findings from three continents. Ophthalmology 108, 697–704 (2001).

    Article  CAS  Google Scholar 

  2. Dithmar, S. et al. Murine high-fat diet and laser photochemical model of basal deposits in Bruch membrane. Arch. Ophthalmol. 119, 1643–1649 (2001).

    Article  CAS  Google Scholar 

  3. Cousins, S.W. et al. The role of aging, high fat diet and blue light exposure in an experimental mouse model for basal laminar deposit formation. Exp. Eye Res. 75, 543–553 (2002).

    Article  CAS  Google Scholar 

  4. Majji, A.B. et al. Age-related retinal pigment epithelium and Bruch's membrane degeneration in senescence-accelerated mouse. Invest. Ophthalmol. Vis. Sci. 41, 3936–3942 (2000).

    CAS  PubMed  Google Scholar 

  5. Weng, J. et al. Insights into the function of Rim protein in photoreceptors and etiology of Stargardt's disease from the phenotype in abcr knockout mice. Cell 98, 13–23 (1999).

    Article  CAS  Google Scholar 

  6. Dithmar, S., Curcio, C.A., Le, N.A., Brown, S. & Grossniklaus, H.E. Ultrastructural changes in Bruch's membrane of apolipoprotein E-deficient mice. Invest. Ophthalmol. Vis. Sci. 41, 2035–2042 (2000).

    CAS  PubMed  Google Scholar 

  7. Rakoczy, P.E. et al. Progressive age-related changes similar to age-related macular degeneration in a transgenic mouse model. Am. J. Pathol. 161, 1515–1524 (2002).

    Article  CAS  Google Scholar 

  8. Lu, B. et al. Abnormalities in monocyte recruitment and cytokine expression in monocyte chemoattractant protein 1-deficient mice. J. Exp. Med. 187, 601–608 (1998).

    Article  CAS  Google Scholar 

  9. Kuziel, W.A. et al. Severe reduction in leukocyte adhesion and monocyte extravasation in mice deficient in CC chemokine receptor 2. Proc. Natl. Acad. Sci. USA 94, 12053–12058 (1997).

    Article  CAS  Google Scholar 

  10. Green, W.R. & Enger, C. Age-related macular degeneration histopathologic studies. The 1992 Lorenz E. Zimmerman Lecture. Ophthalmology 100, 1519–1535 (1993).

    Article  CAS  Google Scholar 

  11. Kamei, M. & Hollyfield, J.G. TIMP-3 in Bruch's membrane: changes during aging and in age-related macular degeneration. Invest. Ophthalmol. Vis. Sci. 40, 2367–2375 (1999).

    CAS  PubMed  Google Scholar 

  12. Delori, F.C., Fleckner, M.R., Goger, D.G., Weiter, J.J. & Dorey, C.K. Autofluorescence distribution associated with drusen in age-related macular degeneration. Invest. Ophthalmol. Vis. Sci. 41, 496–504 (2000).

    CAS  PubMed  Google Scholar 

  13. Eldred, G.E. & Lasky, M.R. Retinal age pigments generated by self-assembling lysosomotropic detergents. Nature 361, 724–726 (1993).

    Article  CAS  Google Scholar 

  14. Suter, M. et al. Age-related macular degeneration. The lipofuscin component N-retinyl-N-retinylidene ethanolamine detaches proapoptotic proteins from mitochondria and induces apoptosis in mammalian retinal pigment epithelial cells. J. Biol. Chem. 275, 39625–39630 (2000).

    Article  CAS  Google Scholar 

  15. Finnemann, S.C., Leung, L.W. & Rodriguez-Boulan, E. The lipofuscin component A2E selectively inhibits phagolysosomal degradation of photoreceptor phospholipid by the retinal pigment epithelium. Proc. Natl. Acad. Sci. USA 99, 3842–3847 (2002).

    Article  CAS  Google Scholar 

  16. Ryan, S.J. Subretinal neovascularization. Natural history of an experimental model. Arch. Ophthalmol. 100, 1804–1809 (1982).

    Article  CAS  Google Scholar 

  17. Tobe, T. et al. Targeted disruption of the FGF2 gene does not prevent choroidal neovascularization in a murine model. Am. J. Pathol. 153, 1641–1646 (1998).

    Article  CAS  Google Scholar 

  18. Spilsbury, K., Garrett, K.L., Shen, W.Y., Constable, I.J. & Rakoczy, P.E. Overexpression of vascular endothelial growth factor (VEGF) in the retinal pigment epithelium leads to the development of choroidal neovascularization. Am. J. Pathol. 157, 135–144 (2000).

    Article  CAS  Google Scholar 

  19. Baffi, J., Byrnes, G., Chan, C.C. & Csaky, K.G. Choroidal neovascularization in the rat induced by adenovirus mediated expression of vascular endothelial growth factor. Invest. Ophthalmol. Vis. Sci. 41, 3582–3589 (2000).

    CAS  PubMed  Google Scholar 

  20. Schwesinger, C. et al. Intrachoroidal neovascularization in transgenic mice overexpressing vascular endothelial growth factor in the retinal pigment epithelium. Am. J. Pathol. 158, 1161–1172 (2001).

    Article  CAS  Google Scholar 

  21. Mullins, R.F., Russell, S.R., Anderson, D.H. & Hageman, G.S. Drusen associated with aging and age-related macular degeneration contain proteins common to extracellular deposits associated with atherosclerosis, elastosis, amyloidosis, and dense deposit disease. FASEB J. 14, 835–846 (2000).

    Article  CAS  Google Scholar 

  22. Johnson, L.V., Ozaki, S., Staples, M.K., Erickson, P.A. & Anderson, D.H. A potential role for immune complex pathogenesis in drusen formation. Exp. Eye Res. 70, 441–449 (2000).

    Article  CAS  Google Scholar 

  23. Anderson, D.H., Mullins, R.F., Hageman, G.S. & Johnson, L.V. A role for local inflammation in the formation of drusen in the aging eye. Am. J. Ophthalmol. 134, 411–431 (2002).

    Article  CAS  Google Scholar 

  24. Ishibashi, T. et al. Advanced glycation end products in age-related macular degeneration. Arch. Ophthalmol. 116, 1629–1632 (1998).

    Article  CAS  Google Scholar 

  25. Schulze, M. et al. Glomerular C3c localization indicates ongoing immune deposit formation and complement activation in experimental glomerulonephritis. Am. J. Pathol. 142, 179–187 (1993).

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Brons, R.H. et al. Detection of immune deposits in skin lesions of patients with Wegener's granulomatosis. Ann. Rheum. Dis. 60, 1097–1102 (2001).

    Article  CAS  Google Scholar 

  27. Bian, Z.M. et al. Glycated serum albumin induces chemokine gene expression in human retinal pigment epithelial cells. J. Leukoc. Biol. 60, 405–414 (1996).

    Article  CAS  Google Scholar 

  28. Duvall, J. & Tso, M.O. Cellular mechanisms of resolution of drusen after laser coagulation. An experimental study. Arch. Ophthalmol. 103, 694–703 (1985).

    Article  CAS  Google Scholar 

  29. Riedemann, N.C. et al. Expression and function of the C5a receptor in rat alveolar epithelial cells. J. Immunol. 168, 1919–1925 (2002).

    Article  CAS  Google Scholar 

  30. Lu, M. et al. Advanced glycation end products increase retinal vascular endothelial growth factor expression. J. Clin. Invest. 101, 1219–1224 (1998).

    Article  CAS  Google Scholar 

  31. Hoffmann, S., Friedrichs, U., Eichler, W., Rosenthal, A. & Wiedemann, P. Advanced glycation end products induce choroidal endothelial cell proliferation, matrix metalloproteinase-2 and VEGF upregulation in vitro. Graefes Arch. Clin. Exp. Ophthalmol. 240, 996–1002 (2002).

    Article  CAS  Google Scholar 

  32. Roberts, W.G. & Palade, G.E. Neovasculature induced by vascular endothelial growth factor is fenestrated. Cancer Res. 57, 765–772 (1997).

    CAS  PubMed  Google Scholar 

  33. Penfold, P.L., Provis, J.M., Furby, J.H., Gatenby, P.A. & Billson, F.A. Autoantibodies to retinal astrocytes associated with age-related macular degeneration. Graefes Arch. Clin. Exp. Ophthalmol. 228, 270–274 (1990).

    Article  CAS  Google Scholar 

  34. Gurne, D.H., Tso, M.O., Edward, D.P. & Ripps, H. Antiretinal antibodies in serum of patients with age-related macular degeneration. Ophthalmology 98, 602–607 (1991).

    Article  CAS  Google Scholar 

  35. Mullins, R.F., Aptsiauri, N. & Hageman, G.S. Structure and composition of drusen associated with glomerulonephritis: implications for the role of complement activation in drusen biogenesis. Eye 15, 390–395 (2001).

    Article  CAS  Google Scholar 

  36. Leys, A. et al. Subretinal neovascular membranes associated with chronic membranoproliferative glomerulonephritis type II. Graefes Arch. Clin. Exp. Ophthalmol. 228, 499–504 (1990).

    Article  CAS  Google Scholar 

  37. Ishida, O. et al. Is Chlamydia pneumoniae infection a risk factor for age related macular degeneration? Br. J. Ophthalmol. 87, 523–524 (2003).

    Article  CAS  Google Scholar 

  38. Kalayoglu, M.V., Galvan, C., Mahdi, O.S., Byrne, G.I. & Mansour, S. Serological association between Chlamydia pneumoniae infection and age-related macular degeneration. Arch. Ophthalmol. 121, 478–482 (2003).

    Article  Google Scholar 

  39. Kothe, H. et al. Hydroxymethylglutaryl coenzyme A reductase inhibitors modify the inflammatory response of human macrophages and endothelial cells infected with Chlamydia pneumoniae. Circulation 101, 1760–1763 (2000).

    Article  CAS  Google Scholar 

  40. Meda, L. et al. β-amyloid (25-35) peptide and interferon-γ synergistically induce the production of the chemotactic cytokine MCP-1/JE in monocytes and microglial cells. J. Immunol. 157, 1213–1218 (1996).

    CAS  PubMed  Google Scholar 

  41. Boring, L., Gosling, J., Cleary, M. & Charo, I.F. Decreased lesion formation in CCR2−/− mice reveals a role for chemokines in the initiation of atherosclerosis. Nature 394, 894–897 (1998).

    Article  CAS  Google Scholar 

  42. Gu, L. et al. Absence of monocyte chemoattractant protein-1 reduces atherosclerosis in low density lipoprotein receptor-deficient mice. Mol. Cell 2, 275–281 (1998).

    Article  CAS  Google Scholar 

  43. Penfold, P.L., Madigan, M.C., Gillies, M.C. & Provis, J.M. Immunological and aetiological aspects of macular degeneration. Prog. Retin. Eye Res. 20, 385–414 (2001).

    Article  CAS  Google Scholar 

  44. Grossniklaus, H.E. et al. Macrophage and retinal pigment epithelium expression of angiogenic cytokines in choroidal neovascularization. Mol. Vis. 8, 119–126 (2002).

    CAS  PubMed  Google Scholar 

  45. Sakurai, E. et al. Targeted disruption of the CD18 or ICAM-1 gene inhibits choroidal neovascularization. Invest. Ophthalmol. Vis. Sci. 44, 2743–2749 (2003).

    Article  Google Scholar 

  46. Sakurai, E., Anand, A., Ambati, B.K., van Rooijen, N. & Ambati, J. Macrophage depletion inhibits experimental choroidal neovascularization. Invest. Ophthalmol. Vis. Sci. 44, 3578–3585 (2003).

    Article  Google Scholar 

  47. Ambati, J., Ambati, B.K., Yoo, S.H., Ianchulev, S. & Adamis, A.P. Age-related macular degeneration: etiology, pathogenesis, and therapeutic strategies. Surv. Ophthalmol. 48, 257–293 (2003).

    Article  Google Scholar 

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Acknowledgements

We thank M.G. Engle, M. Jennes and R. King for assistance with histology and electron microscopy; G. Chen for maintaining cell cultures; M.H. Hanson for photography; J. Husemann for technical advice; H.E. Grossniklaus for expertise in interpreting electron micrographs; and A.P. Adamis, S. Bondada, D.C. Collins, R. Mohan, E.J. Smart, V. Kumar, P.A. Pearson, A.M. Rao, G.S. Rao and J.G. Woodward for valuable discussions. J.A. was supported by a Foundation Fighting Blindness Career Development Award, the Dennis W. Jahnigen Career Development Award administered by the American Geriatrics Society and funded by the John A. Hartford Foundation and Atlantic Philanthropies, grants from Prevent Blindness America and Fight for Sight and a physician-scientist award from University of Kentucky; E.S. was supported by a Fight For Sight postdoctoral fellowship; and B.K.A. was supported by a Knights-Templar Eye Foundation grant.

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Authors and Affiliations

  1. Department of Ophthalmology, University of Kentucky, 740 South Limestone Street, Lexington, 40536-0284, Kentucky, USA

    Jayakrishna Ambati, Akshay Anand & Eiji Sakurai

  2. Department of Microbiology and Immunology, University of Kentucky, MS423 Medical Science Building, Lexington, 40506-0286, Kentucky, USA

    Stefan Fernandez

  3. Department of Chemistry, University of Kentucky, A053 ASTeCC Building, Lexington, 40506-0286, Kentucky, USA

    Bert C Lynn

  4. Institute for Cellular and Molecular Biology, University of Texas, Austin, 78712-1095, Texas, USA

    William A Kuziel

  5. Dana Farber Cancer Institute, Brigham & Women's Hospital, Harvard Medical School, 44 Binney Street Mayer 430, Boston, 02115, Massachusetts, USA

    Barrett J Rollins

  6. Department of Ophthalmology, Medical College of Georgia, Augusta, 30907, Georgia, USA

    Balamurali K Ambati

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Correspondence to Jayakrishna Ambati.

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J.A. is listed on an initial patent filing by the University of Kentucky, describing the findings in these mice.

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Ambati, J., Anand, A., Fernandez, S. et al. An animal model of age-related macular degeneration in senescent Ccl-2- or Ccr-2-deficient mice. Nat Med 9, 1390–1397 (2003). https://doi.org/10.1038/nm950

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