Expression pattern in human macrophages dependent on 9p21.3 coronary artery disease risk locus

Abstract

Objective: Genome-wide association studies identified a risk haplotype on chromosome 9p21.3 to be associated with coronary artery disease (CAD) and myocardial infarction (MI). Since this region does not contain a clear candidate gene with known pathophysiology, we performed a haplotype-specific expression study in human macrophages during pro-inflammatory stimulation to investigate the locus-dependent expression patterns in a model of atherosclerosis, the underlying cause for CAD and MI.

Methods: Blood samples were taken from 40 male stable MI patients either homozygous for 9p21.3 risk (n = 20) or non-risk haplotype (n = 20) as well as from 28 healthy male individuals (n = 14 for each haplotype). Monocytes were isolated by density gradient centrifugation followed by differentiation into macrophages via M-CSF. Macrophages were either incubated with a pro-inflammatory IFNγ-LPS cocktail or kept untreated as controls. After 24 h, RNA was isolated and applied to Affymetrix Human Exon 1.0 ST Arrays.

Results: Macrophages from MI patients and controls stratified for 9p21.3 haplotypes, exhibited marked differences in gene expression. Most pronounced differences were found in inflammatory mediators, like the chemokines CCL8 and CCL2 and the lectines CLEC4E and CLEC5A. Differences in expression changes could be seen most obviously during inflammatory stimulation for both, the interleukins IL12B and IL1B, and members of metallothionein gene family.

Conclusion: These findings show that gene expression is different in 9p21.3 haplotype-stratified macrophages. While these effects are relatively small in our in vitro model of atherosclerosis, these biological effects may contribute to a long term effect in risk haplotype carriers increasing susceptibility to CAD and MI.