Allele-specific NKX2-5 binding underlies multiple genetic associations with human electrocardiographic traits

The cardiac transcription factor (TF) gene NKX2–5 has been associated with electrocardiographic (EKG) traits through GWAS, but the extent to which differential binding of NKX2–5 at common regulatory variants contributes to these traits has not yet been studied. We analyzed transcriptomic and epigenomic data from iPSC-derived cardiomyocytes (iPSC-CMs) from seven related individuals and identified ~2,000 single nucleotide variants (SNVs) associated with allele-specific effects (ASE) on NKX2–5 binding. NKX2–5 ASE-SNVs were enriched for altered TF motifs, for heart-specific eQTLs, and for EKG GWAS signals. Using fine-mapping combined with epigenomic data from iPSC-CMs, we prioritized candidate causal variants for EKG traits, many of which were NKX2–5 ASE-SNVs. Experimentally characterizing two NKX2–5 ASE-SNVs (rs3807989 and rs590041) showed that they modulate the expression of target genes via differential protein binding in cardiac cells, indicating that they are functional variants underlying EKG GWAS signals. Our results show that differential NKX2–5 binding at numerous regulatory variants across the genome contributes to EKG phenotypes.