title: Allele-specific silencing of EEC p63 mutant R304W restores p63 transcriptional activity.
abstract: EEC (ectrodactily-ectodermal dysplasia and cleft lip/palate) syndrome is a rare genetic disease, autosomal dominant inherited. It is part of the ectodermal dysplasia disorders caused by heterozygous mutations in TP63 gene. EEC patients present limb malformations, orofacial clefting, skin and skin's appendages defects, ocular abnormalities. The transcription factor p63, encoded by TP63, is a master gene for the commitment of ectodermal-derived tissues, being expressed in the apical ectodermal ridge is critical for vertebrate limb formation and, at a later stage, for skin and skin's appendages development. The ΔNp63α isoform is predominantly expressed in epithelial cells and it is indispensable for preserving the self-renewal capacity of adult stem cells and to engage specific epithelial differentiation programs. Small interfering RNA (siRNA) offers a potential therapy approach for EEC patients by selectively silencing the mutant allele. Here, using a systemic screening based on a dual-luciferase reported gene assay, we have successfully identified specific siRNAs for repressing the EEC-causing p63 mutant, R304W. Upon siRNA treatment, we were able to restore ΔNp63-WT allele transcriptional function in induced pluripotent stem cells that were derived from EEC patient biopsy. This study demonstrates that siRNAs approach is promising and, may pave the way for curing/delaying major symptoms, such as cornea degeneration and skin erosions in young EEC patients.
authors.affiliation: Department of Experimental Medicine and Surgery, University of Rome 'Tor Vergata', Rome, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Novelli
authors.affiliation: Department of Experimental Medicine and Surgery, University of Rome 'Tor Vergata', Rome, Italy.
authors.forename: A M
authors.initials: AM
authors.lastname: Lena
authors.affiliation: Department of Experimental Medicine and Surgery, University of Rome 'Tor Vergata', Rome, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Panatta
authors.affiliation: Department of Genetics and Developmental Biology, The Ruth and Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.
authors.forename: W
authors.initials: W
authors.lastname: Nasser
authors.affiliation: Department of Genetics and Developmental Biology, The Ruth and Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.
authors.forename: R
authors.initials: R
authors.lastname: Shalom-Feuerstein
authors.affiliation: Department of Experimental Medicine and Surgery, University of Rome 'Tor Vergata', Rome, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Candi
authors.affiliation: Department of Experimental Medicine and Surgery, University of Rome 'Tor Vergata', Rome, Italy.|Medical Research Council, Toxicology Unit, Leicester University, Hodgkin Building, Leicester, UK.
authors.forename: G
authors.initials: G
authors.lastname: Melino
mesh_terms: D000483:Alleles; D001483:Base Sequence; D002971:Cleft Lip / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002972:Cleft Palate / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D004476:Ectodermal Dysplasia / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology; D020868:Gene Silencing; D005799:Genes, Dominant; D017930:Genes, Reporter; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism* / Q000473:pathology; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D058990:Molecular Targeted Therapy; D009154:Mutation; D061251:Primary Cell Culture; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D013577:Syndrome; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D025521:Tumor Suppressor Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
chemical_list: D034741:RNA, Small Interfering; C115051:TP63 protein, human; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D008156:Luciferases