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Pathological Features Of Keratoconus And Fuchs Corneal Dystrophy: A Molecular Insight And Cellular Model 1. Isabella Cheung University of Auckland, Auckland, New Zealand Purpose: Commonly causing severe visual impairment necessitating corneal transplantation, Keratoconus and Fuchs Corneal Dystrophy (FCD) manifest due to stromal thinning and endothelial dysfunction, respectively. However, mechanisms of pathogenesis remain unclear; consequently, therapeutics targeting the underlying causes of these disorders and disease models are currently lacking. With the aim of elucidating and modeling cellular processes mediating pathological and pathogenic changes in Keratoconus and FCD, this study investigated protein expression in affected corneas and the establishment of an ex vivo cellular model of disease. Methods: Epithelial cells, keratocytes and endothelial cells were extracted from dystrophic and non-dystrophic human corneal buttons. Global and specific protein expression in each cell type was characterized using antibody-based labeling techniques and mass spectrometry. A model based on isolated keratocytes cultured on a collagenous substrate was assessed for cell phenotype and function with immunocytochemistry and scrape-loaded dye transfer, respectively. Results: A number of proteins were up- and down-regulated in dystrophic corneas. These were identified as cytokines and biomarkers of apoptosis, pluripotency, inflammation and other cellular processes. Cultured keratocytes expressed biomarkers associated with keratocytes, while those of fibroblastic cells were absent. Gap junction proteins were also present and gap junction-permeable dye was observed in intact keratocytes coupled to injured cells. Conclusions: Anomalies in protein expression in dystrophic corneas may reveal cellular processes implicated in pathology and pathogenesis. This improved understanding of Keratoconus and FCD will refine our preliminary disease model and advance the development of therapeutic agents for these corneal dystrophies. |
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