Ocular surface tissue morphogenesis in normal and disease states revealed by genetically modified mice

Many transgenic and knockout mice exhibit pathogenic processes resembling human ocular surface diseases. Thus, the clinical manifestations of mouse lines can provide clues for identifying heritable human diseases of unknown etiology. However, mouse lines using conventional techniques of transgenesis and gene targeting often exhibit embryonic lethality and congenital defects, which preclude the use of such mouse models to study acquired ocular surface tissue diseases. These difficulties can be in part overcome by preparing mouse lines of inducible transgene expression, tissue-specific gene ablation, and inducible tissue-specific gene ablation. Conditional transgenic mouse lines live normally until administration of doxycycline and hormones that induce expression of the transgene and ablation of gene of interest. Toward this goal, we prepared 2 groups of genetically modified mouse lines: (1) transgenesis using keratocan promoter was used to create Kera-rtTA mice (doxycycline-inducible mice) and Cre-LoxP system (ie, Kera-Cre mice; conditional gene ablation in neural crest cell lineage and adult stromal keratocyte) and Kera-CrePR mice (RU-486 inducible); and (2) knock-in strategies were used to create Krt12-rtTA mice (doxycycline inducible), Krt12-Cre mice (conditional ablation in corneal epithelium), and Krt12rtTA-tet-O-Cre mice (doxycycline-inducible corneal epithelium-specific gene ablation). Using these mouse lines, we showed that transforming growth factor (TGF)-beta2 is essential for eye morphogenesis, TGF-alpha is a morphogen for eyelid formation, and lumican is a matrikine that has multiple regulatory functions on cell activities (eg, migration proliferation and gene expression) besides serving as a regulatory molecule of collagen fibrillogenesis. These mouse lines can also be used as models for development of therapeutic treatment regimens of ocular surface diseases using gene therapy and stem cell strategies.