title: Susceptibility to DNA damage as a molecular mechanism for non-syndromic cleft lip and palate.
abstract: Non-syndromic cleft lip/palate (NSCL/P) is a complex, frequent congenital malformation, determined by the interplay between genetic and environmental factors during embryonic development. Previous findings have appointed an aetiological overlap between NSCL/P and cancer, and alterations in similar biological pathways may underpin both conditions. Here, using a combination of transcriptomic profiling and functional approaches, we report that NSCL/P dental pulp stem cells exhibit dysregulation of a co-expressed gene network mainly associated with DNA double-strand break repair and cell cycle control (p = 2.88×10(-2)-5.02×10(-9)). This network included important genes for these cellular processes, such as BRCA1, RAD51, and MSH2, which are predicted to be regulated by transcription factor E2F1. Functional assays support these findings, revealing that NSCL/P cells accumulate DNA double-strand breaks upon exposure to H2O2. Furthermore, we show that E2f1, Brca1 and Rad51 are co-expressed in the developing embryonic orofacial primordia, and may act as a molecular hub playing a role in lip and palate morphogenesis. In conclusion, we show for the first time that cellular defences against DNA damage may take part in determining the susceptibility to NSCL/P. These results are in accordance with the hypothesis of aetiological overlap between this malformation and cancer, and suggest a new pathogenic mechanism for the disease.
authors.affiliation: Human Genome Research Center, Institute for Biosciences, University of São Paulo, São Paulo, Brazil.
authors.forename: Gerson Shigeru
authors.initials: GS
authors.lastname: Kobayashi
authors.forename: Lucas
authors.initials: L
authors.lastname: Alvizi
authors.forename: Daniele Yumi
authors.initials: DY
authors.lastname: Sunaga
authors.forename: Philippa
authors.initials: P
authors.lastname: Francis-West
authors.forename: Anna
authors.initials: A
authors.lastname: Kuta
authors.forename: Bruno Vinícius Pimenta
authors.initials: BV
authors.lastname: Almada
authors.forename: Simone Gomes
authors.initials: SG
authors.lastname: Ferreira
authors.forename: Leonardo Carmo
authors.initials: LC
authors.lastname: de Andrade-Lima
authors.forename: Daniela Franco
authors.initials: DF
authors.lastname: Bueno
authors.forename: Cássio Eduardo
authors.initials: CE
authors.lastname: Raposo-Amaral
authors.forename: Carlos Frederico
authors.initials: CF
authors.lastname: Menck
authors.forename: Maria Rita
authors.initials: MR
authors.lastname: Passos-Bueno
mesh_terms: D019313:BRCA1 Protein / Q000235:genetics; D002453:Cell Cycle / Q000235:genetics; D002478:Cells, Cultured; D002648:Child; D002971:Cleft Lip / Q000235:genetics*; D002972:Cleft Palate / Q000235:genetics*; D053903:DNA Breaks, Double-Stranded; D017931:DNA Primers / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D003782:Dental Pulp / Q000166:cytology*; D050687:E2F1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D053263:Gene Regulatory Networks / Q000235:genetics*; D006801:Humans; D017403:In Situ Hybridization; D051718:MutS Homolog 2 Protein / Q000235:genetics; D040081:Protein Array Analysis; D051135:Rad51 Recombinase / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000378:metabolism*
chemical_list: D019313:BRCA1 Protein; C492913:BRCA1 protein, human; D017931:DNA Primers; D050687:E2F1 Transcription Factor; C493753:E2F1 protein, human; C495153:RAD51 protein, human; D051135:Rad51 Recombinase; C497172:MSH2 protein, human; D051718:MutS Homolog 2 Protein