ABNORMAL KERATIN-1 AND KERATIN-10 CYTOSKELETON IN CULTURED KERATINOCYTES FROM EPIDERMOLYTIC HYPERKERATOSIS CAUSED BY KERATIN-10 MUTATIONS

Epidermolytic hyperkeratosis is caused by mutations of the diffentiation-specific keratins K1 and K10. These mutations produce a weakened cytoskeleton that is prone to collapse resulting in cell fragility and lysis. In this study we have analyzed cultured keratinocytes from EHK patients bearing (10)R-to-H and 15L-to-S mutations within the 1A segment of the K10 rod domain. Keratinocytes were grown submerged in serum-free medium and induced to differentiate by growing to confluence and increasing the Ca++ concentration in the medium. Cultures were either harvested for mRNA sequence analysis or subjected to immunofluorescence microscopy. Differentiating keratinocytes from these patients were found to express these K10 mutations in their mRNA. Moreover, these cells could be distinguished from normal keratinocytes by their aberrrant morphology. EHK keratinocytes frequently exhibited a collapsed perinuclear network of K1/ K10 filaments and sometimes peripheral granules of K1 and K10 aggregates, reminiscent of the cells of the suprabasal layers in these patients. This report documents the expression of mutant keratin 10 in cultured EHK keratinocytes.