Thy-1 Regulates VEGF-Mediated Choroidal Endothelial Cell Activation and Migration: Implications in Neovascular Age-Related Macular Degeneration

This study addresses the hypothesis that age-related stresses upregulate Thy-1 in choroidal endothelial cells (CECs) and contribute to CEC activation and migration, processes important in choroidal neovascularization (CNV). Measurements were made of Thy-1 protein (Western blot) in CECs and Thy-1 mRN...

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Veröffentlicht in:Investigative ophthalmology & visual science Jg. 57; H. 13; S. 5525 - 5534
Hauptverfasser: Wang, Haibo, Han, Xiaokun, Kunz, Eric, Hartnett, M. Elizabeth
Format: Journal Article
Sprache:Englisch
Veröffentlicht: United States The Association for Research in Vision and Ophthalmology 01.10.2016
Schlagworte:
Adult
Aged
Aged, 80 and over
Animals
Blotting, Western
Cell Movement
Cell Proliferation
Cells, Cultured
Choroid - blood supply
Choroid - metabolism
Choroidal Neovascularization - diagnosis
Choroidal Neovascularization - genetics
Choroidal Neovascularization - metabolism
Disease Models, Animal
Endothelial Cells - metabolism
Endothelial Cells - pathology
Female
Gene Expression Regulation
Humans
Macular Degeneration - diagnosis
Macular Degeneration - genetics
Macular Degeneration - metabolism
Male
Mice
Mice, Inbred C57BL
Middle Aged
Real-Time Polymerase Chain Reaction
Retinal Cell Biology
Retinal Pigment Epithelium - metabolism
Retinal Pigment Epithelium - pathology
RNA, Messenger - genetics
Signal Transduction
Thy-1 Antigens - biosynthesis
Thy-1 Antigens - genetics
Tissue Donors
Up-Regulation
Vascular Endothelial Growth Factor A - metabolism
Young Adult
ISSN:1552-5783, 0146-0404, 1552-5783
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Zusammenfassung:This study addresses the hypothesis that age-related stresses upregulate Thy-1 in choroidal endothelial cells (CECs) and contribute to CEC activation and migration, processes important in choroidal neovascularization (CNV). Measurements were made of Thy-1 protein (Western blot) in CECs and Thy-1 mRNA (real time quantitative PCR) in CECs treated with VEGF, CCL11, or PBS or in RPE/choroids from young or old donors or lasered or nonlasered mice. Immunolabeled Thy-1 in ocular sections was compared from young versus old human donor eyes or those with or without neovascular AMD or from lasered versus nonlasered mice. Choroidal endothelial cells transfected with Thy-1 or control siRNA or pretreated with Thy-1 blocking peptide or control were stimulated with VEGF or 7-ketocholesterol (7-KC). Choroidal endothelial cell migration, proliferation, cytoskeletal stress fibers, Rac1 activation, and phosphorylated VEGF receptor 2 (VEGFR2), integrin β3, and Src were measured. Statistics were performed using ANOVA. Thy-1 was expressed in retinal ganglion cells and in vascular endothelial-cadherin-labeled choroid and localized to human or mouse laser-induced CNV lesions. Thy-1 protein and mRNA were significantly increased in CECs treated with VEGF or CCL11 and in RPE/choroids from aged versus young donor eyes or from lasered mice versus nonlasered controls. Knockdown or inhibition of Thy-1 in CECs significantly reduced VEGF-induced CEC migration and proliferation, stress fiber formation and VEGFR2, Src, integrin β3 and Rac1 activation, and 7-KC-induced Rac1 and Src activation. Thy-1 in CECs regulates VEGF-induced CEC activation and migration and links extracellular 7-KC to intracellular signaling. Future studies elucidating Thy-1 mechanisms in neovascular AMD are warranted.
Bibliographie:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1552-5783
0146-0404
1552-5783
DOI:10.1167/iovs.16-19691