1. APR-246/PRIMA-1(MET) rescues epidermal differentiation in skin keratinocytes derived from EEC syndrome patients with p63 mutations.
- Author
-
Shen J, van den Bogaard EH, Kouwenhoven EN, Bykov VJ, Rinne T, Zhang Q, Tjabringa GS, Gilissen C, van Heeringen SJ, Schalkwijk J, van Bokhoven H, Wiman KG, and Zhou H
- Subjects
- Adult, Base Sequence, Binding Sites genetics, Cell Differentiation drug effects, Cell Differentiation genetics, Cells, Cultured, Cleft Lip genetics, Cleft Lip metabolism, Cleft Palate genetics, Cleft Palate metabolism, Ectodermal Dysplasia genetics, Ectodermal Dysplasia metabolism, Epidermis drug effects, Epidermis metabolism, Epidermis pathology, Female, Humans, Keratinocytes metabolism, Male, Middle Aged, Models, Biological, Mutant Proteins chemistry, Mutant Proteins genetics, Mutant Proteins metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, Transcription Factors chemistry, Transcription Factors metabolism, Transcription, Genetic drug effects, Tumor Suppressor Proteins chemistry, Tumor Suppressor Proteins metabolism, Cleft Lip drug therapy, Cleft Lip pathology, Cleft Palate drug therapy, Cleft Palate pathology, Ectodermal Dysplasia drug therapy, Ectodermal Dysplasia pathology, Keratinocytes drug effects, Keratinocytes pathology, Mutation, Quinuclidines pharmacology, Transcription Factors genetics, Tumor Suppressor Proteins genetics
- Abstract
p53 and p63 share extensive sequence and structure homology. p53 is frequently mutated in cancer, whereas mutations in p63 cause developmental disorders manifested in ectodermal dysplasia, limb defects, and orofacial clefting. We have established primary adult skin keratinocytes from ectrodactyly, ectodermal dysplasia, and cleft lip/palate (EEC) syndrome patients with p63 mutations as an in vitro human model to study the disease mechanism in the skin of EEC patients. We show that these patient keratinocytes cultured either in submerged 2D cultures or in 3D skin equivalents have impaired epidermal differentiation and stratification. Treatment of these patient keratinocytes with the mutant p53-targeting compound APR-246/PRIMA-1(MET) (p53 reactivation and induction of massive apoptosis) that has been successfully tested in a phase I/II clinical trial in cancer patients partially but consistently rescued morphological features and gene expression during epidermal stratification in both 2D and 3D models. This rescue coincides with restoration of p63 target-gene expression. Our data show that EEC patient keratinocytes with p63 mutations can be used for characterization of the abnormal molecular circuitry in patient skin and may open possibilities for the design of novel pharmacological treatment strategies for patients with mutant p63-associated developmental abnormalities.
- Published
- 2013
- Full Text
- View/download PDF