Recent advances in highly multiplexed immunoassays have allowed systematic large-scale measurement of hundreds of plasma proteins in large cohort studies. In combination with genotyping, such studies offer the prospect to 1) identify mechanisms involved with regulation of protein expression in plasma, and 2) determine whether the plasma proteins are likely to be causally implicated in disease. We report here the results of genome-wide association (GWA) studies of 83 proteins considered relevant to cardiovascular disease (CVD), measured in 3,394 individuals with multiple CVD risk factors. We identified 79 genome-wide significant (p<5e-8) association signals, 55 of which replicated at P<0.0007 in separate validation studies (n = 2,639 individuals). Using automated text mining, manual curation, and network-based methods incorporating information on expression quantitative trait loci (eQTL), we propose plausible causal mechanisms for 25 trans-acting loci, including a potential post-translational regulation of stem cell factor by matrix metalloproteinase 9 and receptor-ligand pairs such as RANK-RANK ligand. Using public GWA study data, we further evaluate all 79 loci for their causal effect on coronary artery disease, and highlight several potentially causal associations. Overall, a majority of the plasma proteins studied showed evidence of regulation at the genetic level. Our results enable future studies of the causal architecture of human disease, which in turn should aid discovery of new drug targets. Several proteins that circulate in blood have been linked to cardiovascular disease through the use of classic epidemiology and correlation studies. If individuals with higher risk of disease have higher levels of a protein, the protein may be associated with disease. However, this does not necessarily mean that the protein causes disease; it may merely be an innocent bystander or a consequence of the disease process. To establish whether a protein causes disease, a genetic approach, insensitive to reverse causation, can be used. Instead of correlating the levels of the protein itself, gene variants that regulate the protein levels are used in the analysis. This approach requires prior knowledge of which genetic variants are linked to individual proteins. Therefore we completed a map of how common genetic variants affect the blood concentration levels of 83 proteins that have been implicated in cardiovascular disease. By using this map of cause-to-effect findings, we gained insights into the regulation of a majority of the proteins under study and how they relate to risk of coronary artery disease. This study provides a map of genetic regulation of important cardiovascular plasma proteins, insights into their upstream regulatory environment, as well as novel leads for cardiovascular drug development.

Background— Plasma levels of coagulation factors VII (FVII), VIII (FVIII), and von Willebrand factor (vWF) influence risk of hemorrhage and thrombosis. We conducted genome-wide association studies to identify new loci associated with plasma levels. Methods and Results— The setting of the study included 5 community-based studies for discovery comprising 23 608 European-ancestry participants: Atherosclerosis Risk In Communities Study, Cardiovascular Health Study, British 1958 Birth Cohort, Framingham Heart Study, and Rotterdam Study. All subjects had genome-wide single-nucleotide polymorphism (SNP) scans and at least 1 phenotype measured: FVII activity/antigen, FVIII activity, and vWF antigen. Each study used its genotype data to impute to HapMap SNPs and independently conducted association analyses of hemostasis measures using an additive genetic model. Study findings were combined by meta-analysis. Replication was conducted in 7604 participants not in the discovery cohort. For FVII, 305 SNPs exceeded the genome-wide significance threshold of 5.0×10 ⁻⁸ and comprised 5 loci on 5 chromosomes: 2p23 (smallest P value 6.2×10 ⁻²⁴ ), 4q25 (3.6×10 ⁻¹² ), 11q12 (2.0×10 ⁻¹⁰ ), 13q34 (9.0×10 ⁻²⁵⁹ ), and 20q11.2 (5.7×10 ⁻³⁷ ). Loci were within or near genes, including 4 new candidate genes and F7 (13q34). For vWF, 400 SNPs exceeded the threshold and marked 8 loci on 6 chromosomes: 6q24 (1.2×10 ⁻²² ), 8p21 (1.3×10 ⁻¹⁶ ), 9q34 (<5.0×10 ⁻³²⁴ ), 12p13 (1.7×10 ⁻³² ), 12q23 (7.3×10 ⁻¹⁰ ), 12q24.3 (3.8×10 ⁻¹¹ ), 14q32 (2.3×10 ⁻¹⁰ ), and 19p13.2 (1.3×10 ⁻⁹ ). All loci were within genes, including 6 new candidate genes, as well as ABO (9q34) and VWF (12p13). For FVIII, 5 loci were identified and overlapped vWF findings. Nine of the 10 new findings were replicated. Conclusions— New genetic associations were discovered outside previously known biological pathways and may point to novel prevention and treatment targets of hemostasis disorders.

In this work related to familial aggregation of familial venous thromboembolism, the investigators report genomic and transcriptomic association of 16 novel susceptibility loci for venous thromboembolism.