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

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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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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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