Susceptibility to DNA damage as a molecular mechanism for non-syndromic cleft lip and palate

• Published in: PloS one 2013-06, Vol.8 (6), p.e65677-e65677
• Main Authors: Kobayashi, Gerson Shigeru, Alvizi, Lucas, Sunaga, Daniele Yumi, Francis-West, Philippa, Kuta, Anna, Almada, Bruno Vinícius Pimenta, Ferreira, Simone Gomes, de Andrade-Lima, Leonardo Carmo, Bueno, Daniela Franco, Raposo-Amaral, Cássio Eduardo, Menck, Carlos Frederico, Passos-Bueno, Maria Rita
• Format: Article
• Language: English
• Subjects: Biology; Birth defects; BRCA1 protein; BRCA1 Protein - genetics; Breast cancer; Cancer; Cell adhesion & migration; Cell culture; Cell cycle; Cell Cycle - genetics; Cells, Cultured; Child; Cleft Lip - genetics; Cleft lip/palate; Cleft Palate - genetics; Congenital defects; Defects; Dental pulp; Dental Pulp - cytology; Deoxyribonucleic acid; Disease; DNA; DNA Breaks, Double-Stranded; DNA damage; DNA Primers - genetics; DNA repair; DNA Repair - genetics; Double-strand break repair; E2F1 Transcription Factor - genetics; E2F1 Transcription Factor - metabolism; Embryogenesis; Embryonic development; Embryonic growth stage; Environmental factors; Flow Cytometry; Gene expression; Gene Expression Profiling; Gene Regulatory Networks - genetics; Genes; Genetic aspects; Genetic disorders; Genomes; Humans; Hydrogen peroxide; In Situ Hybridization; Morphogenesis; MSH2 protein; MutS Homolog 2 Protein - genetics; Pathogenesis; Primordia; Protein Array Analysis; Rad51 Recombinase - genetics; Real-Time Polymerase Chain Reaction; Stem cells; Stem Cells - metabolism; Transcription (Genetics)
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0065677

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.