Riudavets Puig, R.; Brorson, I. S.; O'Mahony, D.; Kristensen, V. N.; Mathelier, A.

Transcription is regulated through the sequence-specific binding of transcription factors (TFs) to cis-regulatory regions (CRRs). Although scattered along the genome, multiple CRRs are brought in close spatial proximity to the genes they regulate through the formation of DNA loops. The sequence component of transcriptional regulation suggests that DNA variants in CRRs could disrupt TF-DNA interactions and gene regulatory networks through a cascading effect. To date, only a few cases of recurrent cis-regulatory variants have been described. As an alternative to variant recurrence, some methods utilize genomic annotations that indicate the functional impact (FI) of a variant and the CRRs of each gene to detect the enrichment of high-impact cis-regulatory variants (CRVs). However, the agreement across the different associations and FI scoring methods remains unexplored. This work demonstrates that gene-CRR and FI scoring methods exhibit little consensus, highlighting the impact of the choice of a specific CRR-score combination in a given analysis. In addition, we demonstrate that cancer genes exhibit a higher frequency of FI variants compared to non-cancer genes. Based on this, we developed HiFIseek, a Snakemake pipeline exploring the enrichment of FI variants in associated genomic regions using all region-score combinations and two enrichment detection methods. We apply HiFIseek to detect genes exhibiting high FI CRVs in eight cancer cohorts and a set of breast cancer risk-associated single-nucleotide polymorphisms (SNPs). In the cancer cohorts, we observe a small degree of agreement across CRR-score combinations. Although HiFIseek detected known cancer-related genes with FI CRVs, most of them did not show significant changes in their expression. In the SNPs cohort, HiFIseek found small consensus across CRR-score combinations. However, one of the combinations returned 18 known cancer-related genes, including BRCA1, showing an enrichment of high-impact CRVs.