CREATION OF TRANSGENIC MOUSE MODEL FOR LATTICE AND AVELLINO CORNEAL DYSTROPHIES

Vanderbilt University, Nashville TN

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Abstract

Keratoepithelin (KE) is an extracellular matrix protein encoded by the Big-h3 gene on human chromosome 5q. It is expressed in human cornea, skin and other tissues and its expression is up-regulated with TGFb. KE promotes the adhesion and spreading of dermal fibroblasts in vitro and cell-extracellular matrix interaction in corneal development and wound healing. Recently, DNA point mutations have been identified in KE in patients with a hereditary form of comeal stromal opacities-corneal stromal dystrophies. We postulate that KE plays a universally important role in maintaining corneal optical clarity and in the wound healing of epithelial tissues in cornea and skin. In this study, we plan to create a transgenic mouse model for the human lattice and Avellino corneal dystrophies with KE mutant constructs. We will make DNA constructs, each of which contains the KE mutant cDNA that corresponds to one of the two human corneal dystrophies, lattice (R124C) and Avellino (R124H) respectively. By using cornea-preferred promoters, we will test the KE constructs first in vitro to examine the expression of the KE mutant protein. These KE mutant constructs will then be injected into mouse embryos and the genotypes of these founder pups examined to confirm the presence of the transgenes. Phenotypically, these transgenic mice will be characterized using sit-lamp biomicroscopy to detect the presence of corneal dystrophies, histologic studies of the amyloid studies for the amyloid studies for the amyloid precipitation, and immunohistochemical characterization of the KE deposits. This transgenic mouse model will be valuable in our future wound-healing experiments designed to characterize the biological function of KE, its role in maintaining corneal optical clarity and in wound healing.