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CCR3 is a target for age-related macular degeneration diagnosis and therapy

Abstract

Age-related macular degeneration (AMD), a leading cause of blindness worldwide, is as prevalent as cancer in industrialized nations. Most blindness in AMD results from invasion of the retina by choroidal neovascularisation (CNV). Here we show that the eosinophil/mast cell chemokine receptor CCR3 is specifically expressed in choroidal neovascular endothelial cells in humans with AMD, and that despite the expression of its ligands eotaxin-1, -2 and -3, neither eosinophils nor mast cells are present in human CNV. Genetic or pharmacological targeting of CCR3 or eotaxins inhibited injury-induced CNV in mice. CNV suppression by CCR3 blockade was due to direct inhibition of endothelial cell proliferation, and was uncoupled from inflammation because it occurred in mice lacking eosinophils or mast cells, and was independent of macrophage and neutrophil recruitment. CCR3 blockade was more effective at reducing CNV than vascular endothelial growth factor A (VEGF-A) neutralization, which is in clinical use at present, and, unlike VEGF-A blockade, is not toxic to the mouse retina. In vivo imaging with CCR3-targeting quantum dots located spontaneous CNV invisible to standard fluorescein angiography in mice before retinal invasion. CCR3 targeting might reduce vision loss due to AMD through early detection and therapeutic angioinhibition.

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Figure 1: CCR3 and eotaxins are expressed in CNV.
Figure 2: CCR3 activation promotes angiogenesis.
Figure 3: CNV reduced by CCR3 or eotaxin ablation or blockade independent of leukocyte modulation.
Figure 4: CCR3-targeting quantum dots detect subretinal CNV.

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References

  1. 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 

  2. Gragoudas, E. S., Adamis, A. P., Cunningham, E. T., Feinsod, M. & Guyer, D. R. Pegaptanib for neovascular age-related macular degeneration. N. Engl. J. Med. 351, 2805–2816 (2004)

    Article  CAS  Google Scholar 

  3. Brown, D. M. et al. Ranibizumab versus verteporfin for neovascular age-related macular degeneration. N. Engl. J. Med. 355, 1432–1444 (2006)

    Article  CAS  Google Scholar 

  4. Rosenfeld, P. J. et al. Ranibizumab for neovascular age-related macular degeneration. N. Engl. J. Med. 355, 1419–1431 (2006)

    Article  CAS  Google Scholar 

  5. Famiglietti, E. V. et al. Immunocytochemical localization of vascular endothelial growth factor in neurons and glial cells of human retina. Brain Res. 969, 195–204 (2003)

    Article  CAS  Google Scholar 

  6. Nishijima, K. et al. Vascular endothelial growth factor-A is a survival factor for retinal neurons and a critical neuroprotectant during the adaptive response to ischemic injury. Am. J. Pathol. 171, 53–67 (2007)

    Article  CAS  Google Scholar 

  7. Saint-Geniez, M. et al. Endogenous VEGF is required for visual function: evidence for a survival role on muller cells and photoreceptors. PLoS One 3, e3554 (2008)

    Article  ADS  Google Scholar 

  8. Rothenberg, M. E. & Hogan, S. P. The eosinophil. Annu. Rev. Immunol. 24, 147–174 (2006)

    Article  CAS  Google Scholar 

  9. Submacular Surgery Trials Research Group Histopathologic and ultrastructural features of surgically excised subfoveal choroidal neovascular lesions: submacular surgery trials report no. 7. Arch. Ophthalmol. 123, 914–921 (2005)

    Article  Google Scholar 

  10. Justice, J. P. et al. Ablation of eosinophils leads to a reduction of allergen-induced pulmonary pathology. Am. J. Physiol. Lung Cell. Mol. Physiol. 284, L169–L178 (2003)

    Article  CAS  Google Scholar 

  11. Humbles, A. A. et al. A critical role for eosinophils in allergic airways remodeling. Science 305, 1776–1779 (2004)

    Article  ADS  CAS  Google Scholar 

  12. Pope, S. M., Zimmermann, N., Stringer, K. F., Karow, M. L. & Rothenberg, M. E. The eotaxin chemokines and CCR3 are fundamental regulators of allergen-induced pulmonary eosinophilia. J. Immunol. 175, 5341–5350 (2005)

    Article  CAS  Google Scholar 

  13. Jose, P. J. et al. Eotaxin: a potent eosinophil chemoattractant cytokine detected in a guinea pig model of allergic airways inflammation. J. Exp. Med. 179, 881–887 (1994)

    Article  CAS  Google Scholar 

  14. Teixeira, M. M. et al. Chemokine-induced eosinophil recruitment. Evidence of a role for endogenous eotaxin in an in vivo allergy model in mouse skin. J. Clin. Invest. 100, 1657–1666 (1997)

    Article  CAS  Google Scholar 

  15. Blanchard, C. et al. Eotaxin-3 and a uniquely conserved gene-expression profile in eosinophilic esophagitis. J. Clin. Invest. 116, 536–547 (2006)

    Article  CAS  Google Scholar 

  16. Salcedo, R. et al. Eotaxin (CCL11) induces in vivo angiogenic responses by human CCR3+ endothelial cells. J. Immunol. 166, 7571–7578 (2001)

    Article  CAS  Google Scholar 

  17. Puxeddu, I. et al. Human peripheral blood eosinophils induce angiogenesis. Int. J. Biochem. Cell Biol. 37, 628–636 (2005)

    Article  CAS  Google Scholar 

  18. Heissig, B. et al. Low-dose irradiation promotes tissue revascularization through VEGF release from mast cells and MMP-9-mediated progenitor cell mobilization. J. Exp. Med. 202, 739–750 (2005)

    Article  CAS  Google Scholar 

  19. 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 

  20. Nozaki, M. et al. Drusen complement components C3a and C5a promote choroidal neovascularization. Proc. Natl Acad. Sci. USA 103, 2328–2333 (2006)

    Article  ADS  CAS  Google Scholar 

  21. Nozaki, M. et al. Loss of SPARC-mediated VEGFR-1 suppression after injury reveals a novel antiangiogenic activity of VEGF-A. J. Clin. Invest. 116, 422–429 (2006)

    Article  CAS  Google Scholar 

  22. Kleinman, M. E. et al. Sequence- and target-independent angiogenesis suppression by siRNA via TLR3. Nature 452, 591–597 (2008)

    Article  ADS  CAS  Google Scholar 

  23. 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 

  24. 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 

  25. Humbles, A. A. et al. The murine CCR3 receptor regulates both the role of eosinophils and mast cells in allergen-induced airway inflammation and hyperresponsiveness. Proc. Natl Acad. Sci. USA 99, 1479–1484 (2002)

    Article  ADS  CAS  Google Scholar 

  26. Rothenberg, M. E., MacLean, J. A., Pearlman, E., Luster, A. D. & Leder, P. Targeted disruption of the chemokine eotaxin partially reduces antigen-induced tissue eosinophilia. J. Exp. Med. 185, 785–790 (1997)

    Article  CAS  Google Scholar 

  27. Kitamura, Y., Go, S. & Hatanaka, K. Decrease of mast cells in W/Wv mice and their increase by bone marrow transplantation. Blood 52, 447–452 (1978)

    CAS  PubMed  Google Scholar 

  28. Zhou, J. et al. Neutrophils promote experimental choroidal neovascularization. Mol. Vis. 11, 414–424 (2005)

    CAS  PubMed  Google Scholar 

  29. Fulkerson, P. C. et al. Negative regulation of eosinophil recruitment to the lung by the chemokine monokine induced by IFN-γ (Mig, CXCL9). Proc. Natl Acad. Sci. USA 101, 1987–1992 (2004)

    Article  ADS  CAS  Google Scholar 

  30. Fulkerson, P. C., Zhu, H., Williams, D. A., Zimmermann, N. & Rothenberg, M. E. CXCL9 inhibits eosinophil responses by a CCR3- and Rac2-dependent mechanism. Blood 106, 436–443 (2005)

    Article  CAS  Google Scholar 

  31. Green, W. R. & Key, S. N. Senile macular degeneration: a histopathologic study. Trans. Am. Ophthalmol. Soc. 75, 180–254 (1977)

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Ambati, J. et al. An animal model of age-related macular degeneration in senescent Ccl-2- or Ccr-2-deficient mice. Nature Med. 9, 1390–1397 (2003)

    Article  CAS  Google Scholar 

  33. Ip, M. S. et al. Anti-vascular endothelial growth factor pharmacotherapy for age-related macular degeneration: a report by the American Academy of Ophthalmology. Ophthalmology 115, 1837–1846 (2008)

    Article  Google Scholar 

  34. Sayanagi, K., Sharma, S. & Kaiser, P. K. Photoreceptor status after anti-vascular endothelial growth factor therapy in exudative age-related macular degeneration. Br. J. Ophthalmol. 93, 622–626 (2009)

    Article  CAS  Google Scholar 

  35. Yodoi, Y. et al. Central retinal sensitivity after intravitreal injection of bevacizumab for myopic choroidal neovascularization. Am. J. Ophthalmol. 147, 816–824 (2009)

    Article  CAS  Google Scholar 

  36. Carmeliet, P. et al. Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature 380, 435–439 (1996)

    Article  ADS  CAS  Google Scholar 

  37. Ferrara, N. et al. Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene. Nature 380, 439–442 (1996)

    Article  ADS  CAS  Google Scholar 

  38. Marneros, A. G. et al. Vascular endothelial growth factor expression in the retinal pigment epithelium is essential for choriocapillaris development and visual function. Am. J. Pathol. 167, 1451–1459 (2005)

    Article  CAS  Google Scholar 

  39. Bressler, S. B., Maguire, M. G., Bressler, N. M. & Fine, S. L. (The Macular Photocoagulation Study Group). Relationship of drusen and abnormalities of the retinal pigment epithelium to the prognosis of neovascular macular degeneration. Arch. Ophthalmol. 108, 1442–1447 (1990)

    Article  CAS  Google Scholar 

  40. Macular Photocoagulation Study Group Risk factors for choroidal neovascularization in the second eye of patients with juxtafoveal or subfoveal choroidal neovascularization secondary to age-related macular degeneration. Arch. Ophthalmol. 115, 741–747 (1997)

    Article  Google Scholar 

  41. St Croix, B. et al. Genes expressed in human tumor endothelium. Science 289, 1197–1202 (2000)

    Article  ADS  CAS  Google Scholar 

  42. Zhang, L. et al. Gene expression profiles in normal and cancer cells. Science 276, 1268–1272 (1997)

    Article  CAS  Google Scholar 

  43. Wenzel, S. E. Eosinophils in asthma—closing the loop or opening the door? N. Engl. J. Med. 360, 1026–1028 (2009)

    Article  CAS  Google Scholar 

  44. Geisen, P., McColm, J. R. & Hartnett, M. E. Choroidal endothelial cells transmigrate across the retinal pigment epithelium but do not proliferate in response to soluble vascular endothelial growth factor. Exp. Eye Res. 82, 608–619 (2006)

    Article  CAS  Google Scholar 

  45. Peterson, L. J., Wittchen, E. S., Geisen, P., Burridge, K. & Hartnett, M. E. Heterotypic RPE-choroidal endothelial cell contact increases choroidal endothelial cell transmigration via PI 3-kinase and Rac1. Exp. Eye Res. 84, 737–744 (2007)

    Article  CAS  Google Scholar 

  46. Smith, J. R. et al. Unique gene expression profiles of donor-matched human retinal and choroidal vascular endothelial cells. Invest. Ophthalmol. Vis. Sci. 48, 2676–2684 (2007)

    Article  Google Scholar 

  47. Zamora, D. O. et al. Proteomic profiling of human retinal and choroidal endothelial cells reveals molecular heterogeneity related to tissue of origin. Mol. Vis. 13, 2058–2065 (2007)

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank R. King, L. Xu, M. McConnell, K. Emerson, G. R. Pattison and M. Mingler for technical assistance, J. M. Farber for the gift of a reagent, R. J. Kryscio for statistical guidance, and B. Appukuttan, M. W. Fannon, R. Mohan, A. P. Pearson, A. M. Rao, G. S. Rao and K. Ambati for discussions. J.A. was supported by National Eye Institute/National Institutes of Health (NIH) grants EY015422, EY018350 and EY018836, the Doris Duke Distinguished Clinical Scientist Award, the Burroughs Wellcome Fund Clinical Scientist Award in Translational Research, the Macula Vision Research Foundation, the E. Matilda Ziegler Foundation for the Blind, the Dr. E. Vernon Smith and Eloise C. Smith Macular Degeneration Endowed Chair, the Lew R. Wassermann Merit & Physician Scientist Awards (Research to Prevent Blindness, RPB), the American Health Assistance Foundation, and a departmental unrestricted grant from the RPB. J.Z.B. was supported by the University of Kentucky Physician Scientist Award. M.E.K. was supported by the International Retinal Research Foundation Dr. Charles Kelman Postdoctoral Scholar Award. R.J.C.A. was supported by Fight for Sight. B.K.A. was supported by NIH grants EY017182 and EY017950, the VA Merit Award and the Department of Defense. M.E.R. was supported by NIH grants AI45898 and DK076893. C.J.G. was supported by NIH grant AI039759. M.E.H. was supported by NIH grants EY017011 and EY015130 and a RPB departmental unrestricted grant. J.R.S. was supported by NIH grant EY010572, and RPB Career Development Award and a departmental unrestricted grant.

Author Contributions A.T., J.Z.B., M.E.K., W.G.C., M.N., K.Y., H.K., R.J.C.A., S.D., K.S., B.J.R., M.G.G., S.J.B., P.G. and A.M. performed experiments. S.G., A.A.H., Y.P., J.D.W., J.R.S., Y.O. and T.I. provided reagents. J.A. conceived and directed the project, and, with assistance from B.K.A., M.E.H., M.E.R., R.J.C.A. and J.R.S., wrote the paper. All authors had the opportunity to discuss the results and comment on the manuscript.

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Correspondence to Jayakrishna Ambati.

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J.A. and M.E.K. are named as inventors in a patent application filed by the University of Kentucky on the intellectual property presented in this Article.

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Takeda, A., Baffi, J., Kleinman, M. et al. CCR3 is a target for age-related macular degeneration diagnosis and therapy. Nature 460, 225–230 (2009). https://doi.org/10.1038/nature08151

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