Sequencing of candidate genes in Dominican families implicates both rare exonic and common non-exonic variants for carotid intima-media thickness at bifurcation
BACKGROUND: Through linkage and tagSNP-based association studies in 100 Dominican Republic (DR) families, we previously identified ANLN and AOAH (7p14.3) as candidate genes for carotid intima-media thickness at bifurcation (bIMT). METHODS AND RESULTS: Introns, exons and flanking regions of ANLN and AOAH were re-sequenced in 151 individuals from 9 families with evidence for linkage at 7p14.3. For common variants [CV, minor allele frequency (MAF) ≥ 5%], single variant-based analysis was performed. For rare variants (RV, MAF<5%), gene-based analysis aggregating all RVs within a gene was performed. CV analysis revealed the strongest signal at rs3815483 (P=0.0003) in ANLN and rs60023210 (P=0.00005) in AOAH. In ANLN, RV analysis found suggestive evidence for association with exonic RVs (P=0.08), and in particular non-synonymous RVs (P=0.04) but not with all RVs (P=0.15). The variant alleles of all non-synonymous RVs segregated with the major allele of rs3815483 and were associated with lower bIMT while a novel synonymous RV segregated with the minor allele of rs3815483 and was associated with greater bIMT. Additional analysis in 561 DR individuals found suggestive evidence for association with all ANLN non-synonymous RVs (P=0.08). In AOAH, no evidence for association with RVs was detected. Instead, conditional analysis revealed that multiple independent intronic CVs are associated with bIMT in addition to rs60023210. CONCLUSIONS: We demonstrate the utility of using family-based studies to evaluate the contribution of RVs. Our data suggest two modes of genetic architecture underlying the linkage and association at ANLN (multiple exonic RVs) and AOAH (multiple intronic CVs with uncharacterized functions)
