Upregulation of CCR3 by Age-Related Stresses Promotes Choroidal Endothelial Cell Migration via VEGF-Dependent and -Independent Signaling
Open Access
- 1 October 2011
- journal article
- research article
- Published by Association for Research in Vision and Ophthalmology (ARVO) in Investigative Ophthalmology & Visual Science
- Vol. 52 (11), 8271-8277
- https://doi.org/10.1167/iovs.11-8230
Abstract
Purpose.: To explore the molecular mechanisms by which the C–C chemokine receptor type 3 (CCR3) and chemokine (C–C motif) ligand 11 (CCL11) regulate choroidal endothelial cell (CEC) migration and the interactions with the vascular endothelial growth factor (VEGF) signaling pathway. Methods.: Human retinal sections from young and aged donor normal eyes were immunolabeled. By real-time PCR, CCR3 mRNA was measured in retinal pigmented epithelium (RPE)/choroids obtained from young and aged human donor eyes and in cultured CECs exposed to hydrogen peroxide. CCR3 ligand and CCL11- or VEGF-stimulated CEC migration was also measured in the presence of the CCR3 inhibitor or control using fluorescence microscopy. Activation of Rac1, phosphorylated Akt as a readout for phosphoinositol 3-kinase signaling, and VEGFR2 activation were measured in CECs incubated with CCL11, VEGF, or combined CCL11/VEGF. Results.: CCR3 was expressed to a greater level in older compared with that in younger human retinas or RPE/choroids. Ligand-activated CCR3 increased CEC migration, which was inhibited by the CCR3 inhibitor. Rac1 activity, p-Akt, and p-VEGFR2 were significantly increased in CECs incubated with CCL11. The CCR3 inhibitor prevented VEGF-induced CEC migration and Rac1 activation in CECs. Rac1 activity was additively increased in CECs treated with CCL11 and VEGF compared with that in cells with CCL11 or VEGF treatment alone. Ligand-activated CCR3 caused VEGFR2 phosphorylation and coimmunoprecipitation of VEGFR2 and CCR3. Conclusions.: Activated CCR3 promotes CEC migration and Rac1 activation and causes an association with and activation of VEGFR2. Cross-talk between CCR3 and VEGF signaling exists and may be important in choroidal neovascularization in human age-related macular degeneration.This publication has 31 references indexed in Scilit:
- Localization of collagen XVIII and the endostatin portion of collagen XVIII in aged human control eyes and eyes with age-related macular degeneration.Investigative Ophthalmology & Visual Science, 2004
- Rho and Rac Take Center StageCell, 2004
- Cell Migration: Integrating Signals from Front to BackScience, 2003
- Roles of Rho-family GTPases in cell polarisation and directional migrationCurrent Opinion in Cell Biology, 2003
- Drusen proteome analysis: An approach to the etiology of age-related macular degenerationProceedings of the National Academy of Sciences of the United States of America, 2002
- Activation of Rac and Cdc42 Video Imaged by Fluorescent Resonance Energy Transfer-Based Single-Molecule Probes in the Membrane of Living CellsMolecular and Cellular Biology, 2002
- A Randomized, Placebo-Controlled, Clinical Trial of High-Dose Supplementation With Vitamins C and E, Beta Carotene, and Zinc for Age-Related Macular Degeneration and Vision LossAmerican Journal of Ophthalmology, 2001
- Rho Family GTPases Regulate VEGF-Stimulated Endothelial Cell MotilityExperimental Cell Research, 2001
- Eotaxin (CCL11) Induces In Vivo Angiogenic Responses by Human CCR3+ Endothelial CellsPublished by The American Association of Immunologists ,2001
- Risk factors for age-related macular degeneration: Pooled findings from three continentsOphthalmology, 2001