Allele-specific transcription factor binding in a cellular model of orofacial clefting

• Published in: Scientific reports Vol. 12; no. 1; pp. 1807 - 10
• Main Authors: Ruff, Katharina L M, Hollstein, Ronja, Fazaal, Julia, Thieme, Frederic, Gehlen, Jan, Mangold, Elisabeth, Knapp, Michael, Welzenbach, Julia, Ludwig, Kerstin U
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 02.02.2022; Nature Publishing Group UK; Nature Portfolio
• Subjects: Alleles; AP-2 protein; Binding Sites; Cell Line; Cell lines; Chromatin Immunoprecipitation Sequencing; Cleft Lip - genetics; Cleft Lip - metabolism; Cleft lip/palate; Cleft Palate - genetics; Cleft Palate - metabolism; Congenital defects; Craniofacial growth; Etiology; Gene Expression Regulation, Developmental; Genetic diversity; Genetic Predisposition to Disease; Genetic variance; Genome-wide association studies; Genome-Wide Association Study; Genomes; Human Embryonic Stem Cells - metabolism; Humans; Integration; Localization; Mesenchymal Stem Cells - metabolism; Mesenchyme; Polymorphism, Single Nucleotide; Protein Binding; RNA-Seq; Transcription Factor AP-2 - genetics; Transcription Factor AP-2 - metabolism; Transcription factors; Transcriptome
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-022-05876-7

Non-syndromic cleft lip with/without cleft palate (nsCL/P) is a frequent congenital malformation with multifactorial etiology. While recent genome-wide association studies (GWAS) have identified several nsCL/P risk loci, the functional effects of the associated non-coding variants are largely unknown. Furthermore, additional risk loci remain undetected due to lack of power. As genetic variants might alter binding of transcription factors (TF), we here hypothesized that the integration of data from TF binding sites, expression analyses and nsCL/P GWAS might help to (i) identify functionally relevant variants at GWAS loci, and (ii) highlight novel risk variants that have been previously undetected. Analysing the craniofacial TF TFAP2A in human embryonic palatal mesenchyme (HEPM) cells, we identified 2845 TFAP2A ChIP-seq peaks, several of which were located near nsCL/P candidate genes (e.g. MSX1 and SPRY2). Comparison with independent data suggest that 802 of them might be specific to craniofacial development, and genes near these peaks are enriched in processes relevant to nsCL/P. Integration with nsCL/P GWAS data, however, did not show robust evidence for co-localization of common nsCL/P risk variants with TFAP2A ChIP-seq peaks. This data set represents a new resource for the analyses of craniofacial processes, and similar approaches with additional cell lines and TFs could be applied to generate further insights into nsCL/P etiology.