Extended Data Fig. 1: GWAS identify TIE2 (TEK) as a risk locus for CAD.

a. A LocusZoom plot with eQTL association in GTEX v8 Tibial Artery (top) and CAD GWAS association (middle) and LD matrix (bottom) generated using ezQTL tool. The red triangle indicates the rs1322052 SNP which demonstrates strong colocalization between eQTL and GWAS signals within the LD block (HyPrColoc posterior probability, PP: 0.8789). The insert in top figure indicates the direction of eQTL association with the risk variant T being associated with lower TIE2 expression. b. IGV⁸⁰ genome browser screenshots of the TIE2 locus that demonstrates ATAC-Seq, H3K27ac ChIP-Seq and Hi-C data from teloHAECs treated with 0–24 TNFα stimulus⁶⁵. The rs1322052 localizes to cis-regulatory enhancer element that is TNFα-responsive and loops to the TIE2 promoter. c. CRISPR deletion (CRISPRΔ) and CRISPR inhibition (CRISPRi) experiments were used to study the effect of rs1322052 variant site on TIE2 expression in teloHAECs. Two pairs of gRNAs were used to make the deletion of 945 bps and 1120 bps centered around rs1322052 variant (schematic). Quantification of DNA band intensities on the gel image demonstrate ~80% deletion of the intronic rs1322052 variant (bar plot on the left). The deletion translated to 35–44% reduction in the expression of TIE2 (TEK) gene (bar plot in the middle). CRISPRi-mediated inhibition of the variants site led to ~30% repression of TIE2 expression (bar plot in the right). Data represented as means ± s.d. (for CRISPRΔ, n = 5 independent samples per group, one-way ANOVA (P = 0.0024) with Dunnett’s post hoc test (*P < 0.05 and **P < 0.01); for CRISPRi, n = 3 independent samples per group, two-tailed t-test, unpaired P = 0.0379).