Copy number variation analysis in bicuspid aortic valve-related aortopathy identifies TBX20 as a contributing gene

摘要

Flowchart of our strategy and tools used to identify causative CNVs for BAV/TAA. Genome-wide raw microarray data of 95 unrelated BAV/TAA patients were analysed with GenomeStudio software and an in-house developed online tool called the CNV-Webstore. Rare and unique CNVs with protein-coding genes with a potential role in the cardiovascular system and with a low frequency in healthy individuals (MAF < 1%) were prioritised using data of control samples, NCBI gene, OMIM, PubMed, Human Protein Atlas, Zebrafish Information Network and Mouse Genome Informatics. Only validated CNVs were further investigated. Additional genetic evidence was searched by looking for overlapping CNVs in similar cases in DECIPHER and in-house WES (whole-exome sequencing) data of 67 BAV/TAA patients on which CNV-calling was performed (unpublished data). For CNVs not affecting our gene of interest, public available Hi-C data were consulted for the presence of a potential topological associated domain (TAD), minor allele frequency (MAF) boundary (promoter.bx.psu.edu/hi-c/). In case of supportive evidence, more genetic data for the involvement of the gene of interest in the BAV/TAA pathology by means of a variant burden analysis on rare deleterious next-generation sequencing (NGS) variants detected in 637 BAV/TAA patients and the gnomAD database. BAV bicuspid aortic valve, TAA thoracic aortic aneurysm, SNP single-nucleotide polymorphism, CNV copy number variation, MLPA multiplex ligation-dependent probe amplification, MAQ multiplex amplicon quantification, qPCR quantitative polymerase chain reaction, NCBI National Center for Biotechnology Information, OMIM Online Mendelian Inheritance in Man, TAD topological associated domain, MAF minor allele frequency