Altered Protein Function Caused by AMD-associated Variant rs704 Links Vitronectin to Disease Pathology
| Autor: | Biasella, Fabiola, Plössl, Karolina, Karl, Claudia, Weber, Bernhard H. F., Friedrich, Ulrike |
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| Rok vydání: | 2020 |
| Předmět: |
VTN
Blotting Western 610 Medizin Fluorescent Antibody Technique Retinal Pigment Epithelium AMD AMD age-related macular degeneration vitronectin VTN rs704 vitronectin Retina Macular Degeneration Human Umbilical Vein Endothelial Cells rs704 Humans Protein Isoforms Cloning Molecular age-related macular degeneration ddc:610 Biochemistry and Molecular Biology Genetic Variation Cell Encapsulation Recombinant Proteins eye diseases Extracellular Matrix body regions Electrophoresis Polyacrylamide Gel sense organs |
| Zdroj: | Investigative Ophthalmology & Visual Science |
| DOI: | 10.5283/epub.44277 |
| Popis: | PURPOSE. Vitronectin, a cell adhesion and spreading factor, is suspected to play a role in the pathogenesis of age-related macular degeneration (AMD), as it is a major component of AMD-specific extracellular deposits (e.g., soft drusen, subretinal drusenoid deposits). The present study addressed the impact of AMD-associated non-synonymous variant rs704 in the vitronectin-encoding gene VTN on vitronectin functionality. METHODS. Effects of rs704 on vitronectin expression and processing were analyzed by semi-quantitative sequencing of VTN transcripts from retinal pigment epithelium (RPE) cells generated from human induced pluripotent stem cells (hiPSCs) and from human neural retina, as well as by western blot analyses on heterologously expressed vitronectin isoforms. Binding of vitronectin isoforms to retinal and endothelial cells was analyzed by western blot. Immunofluorescence staining followed extracellular matrix (ECM) deposition in cultured RPE cells heterologously expressing the vitronectin isoforms. Adhesion of fluorescently labeled RPE or endothelial cells in dependence of recombinant vitronectin or vitronectin-containing ECM was investigated fluorometrically or microscopically. Tube formation and migration assays addressed effects of vitronectin on angiogenesis-related processes. RESULTS. Variant rs704 affected expression, secretion, and processing but not oligomerization of vitronectin. Cell binding and influence on RPE-mediated ECM deposition differed between AMD-risk-associated and non-AMD-risk-associated protein isoforms. Finally, vitronectin affected adhesion and endothelial tube formation. CONCLUSIONS. The AMD-risk-associated vitronectin isoform exhibits increased expression and altered functionality in cellular processes related to the sub-RPE aspects of AMD pathology. Although further research is required to address the subretinal disease aspects, this initial study supports an involvement of vitronectin in AMD pathogenesis. |
| Databáze: | OpenAIRE |
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