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Mariya Ali, Jian Hua Qi, Alyson Wolk, Alecia Cutler, Heidi Stohr, Courtney Hershberger, Gautam Mahajan, Chandrasekhar Kothapalli, Bela Anand-Apte; Alterations in microvilli on the surface of retinal Pigment Epithelial (RPE) cells in Sorsby’s Fundus Dystrophy. Invest. Ophthalmol. Vis. Sci. 2019;60(9):1913.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Sorsby’s Fundus Dystrophy (SFD) is an autosomal dominant macular disorder that is caused by mutations in the tissue inhibitor of metalloproteinase-3 (TIMP3) gene. Histopathological analyses reveal the presence of large amorphous lipid enriched extracellular deposits located between the basal lamina of the retinal pigment epithelium and the inner collagenous layer of Bruch’s membrane that are indicative of abnormal RPE function. The purpose of this study was to evaluate the consequences of Ser179Cys-TIMP3 mutation on the cellular morphology and function of the RPE and its extracellular matrix to provide insights into the pathogenesis of the disease.
Methods: RPE was isolated from homozygous TIMP3S179C/S179C mice and their littermate WT controls. RNA was extracted and used for RNA sequencing analysis. Gene set enrichment analysis (GSEA) was performed using hallmark and gene ontology gene sets. For in vitro studies ARPE-19 cells that stably express Ser179Cys-TIMP3 were cultured on trans-well inserts for one to three months. Morphology of the cells was evaluated by scanning electron microscopy (SEM) and immunofluorescence. The extracellular matrix on the basal surface was observed by decellularizing the trans-well mesh, followed by SEM.
Results: GSEA analysis of RNA sequencing data determined that genes involved in apical surface (from Hallmark gene set) and various extracellular components (Gene Ontology gene sets) were enriched in the RPE of TIMP3S179C/S179C mice. SEM of the apical surface of ARPE-19 cells expressing Ser179Cys-TIMP3 showed decreased microvilli and altered extracellular matrix.
Conclusions: Changes to the morphology and extracellular matrix of the RPE may reveal key insights to disease pathology. These results may lead to new insights for disease pathogenesis in macular dystrophies.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
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