APR-246/PRIMA-1(MET) rescues epidermal differentiation in skin keratinocytes derived from EEC syndrome patients with p63 mutations.
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.
Department of Oncology-Pathology, Cancer Center Karolinska,  Karolinska Institutet, SE-171 76 Stockholm,  Sweden.
Jinfeng
J
Shen
Ellen H
EH
van den Bogaard
Evelyn N
EN
Kouwenhoven
Vladimir J N
VJ
Bykov
Tuula
T
Rinne
Qiang
Q
Zhang
Geuranne S
GS
Tjabringa
Christian
C
Gilissen
Simon J
SJ
van Heeringen
Joost
J
Schalkwijk
Hans
H
van Bokhoven
Klas G
KG
Wiman
Huiqing
H
Zhou
D000328:Adult; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D002971:Cleft Lip / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002972:Cleft Palate / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D004476:Ectodermal Dysplasia / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D004817:Epidermis / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005260:Female; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D011812:Quinuclidines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D025521:Tumor Suppressor Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D050505:Mutant Proteins; D011812:Quinuclidines; D012333:RNA, Messenger; C115051:TP63 protein, human; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C533410:eprenetapopt