APR-246/PRIMA-1(MET) rescues epidermal differentiation in skin keratinocytes derived from EEC syndrome patients with p63 mutations

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 110; no. 6; p. 2157
• Main Authors: Shen, Jinfeng, van den Bogaard, Ellen H, Kouwenhoven, Evelyn N, Bykov, Vladimir J N, Rinne, Tuula, Zhang, Qiang, Tjabringa, Geuranne S, Gilissen, Christian, van Heeringen, Simon J, Schalkwijk, Joost, van Bokhoven, Hans, Wiman, Klas G, Zhou, Huiqing
• Format: Journal Article
• Language: English
• Published: United States 05.02.2013
• Subjects: Adult; Base Sequence; Binding Sites - genetics; Cell Differentiation - drug effects; Cell Differentiation - genetics; Cells, Cultured; Cleft Lip - drug therapy; Cleft Lip - genetics; Cleft Lip - metabolism; Cleft Lip - pathology; Cleft Palate - drug therapy; Cleft Palate - genetics; Cleft Palate - metabolism; Cleft Palate - pathology; Ectodermal Dysplasia - drug therapy; Ectodermal Dysplasia - genetics; Ectodermal Dysplasia - metabolism; Ectodermal Dysplasia - pathology; Epidermis - drug effects; Epidermis - metabolism; Epidermis - pathology; Female; Humans; Keratinocytes - drug effects; Keratinocytes - metabolism; Keratinocytes - pathology; Male; Medicin och hälsovetenskap; Middle Aged; Models, Biological; Mutant Proteins - chemistry; Mutant Proteins - genetics; Mutant Proteins - metabolism; Mutation; Quinuclidines - pharmacology; RNA, Messenger - genetics; RNA, Messenger - metabolism; Transcription Factors - chemistry; Transcription Factors - genetics; Transcription Factors - metabolism; Transcription, Genetic - drug effects; Tumor Suppressor Proteins - chemistry; Tumor Suppressor Proteins - genetics; Tumor Suppressor Proteins - metabolism
• ISSN: 1091-6490; 1091-6490
• DOI: 10.1073/pnas.1201993110

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.