Inflammation-Induced Alternative Splicing in Human Endothelial Cells Reveals Genetic Mechanisms of Cardiovascular Disease Risk

Abstract

Alternative splicing modulates mRNA protein-coding sequence, stability, and translation rates, although it has not been comprehensively annotated in human endothelial cells (ECs). EC dysfunction is a hallmark of complex inflammatory diseases, including cancer and atherosclerosis. Therefore, this study modeled acute inflammation in vitro using 53 genetically distinct human aortic EC lines exposed to interleukin-1{beta} (IL-1{beta}) or control media. This approach identified 1,224 differentially spliced transcripts (DSTs) between IL-1{beta} and control conditions. DSTs were enriched for alternative first (AF) exons, including several novel mRNA isoforms of disease-associated and metabolic genes. It was hypothesized and confirmed that AF splicing was driven by alternative promoters using ATAC-seq and ChIP-seq data. To identify alternative promoters driving IL-1{beta}-dependent AF isoforms, a quantitative measure of promoter activity ratios was defined, and analysis found that histone 3 lysine 27 acetylation and binding of the transcription factors ERG and RELA often correlated with alternative promoter usage. Finally, the effect of common genetic variants on alternative first exon usage was interrogated through splicing quantitative trait locus (sQTL) analysis. Significant sQTLs were next submitted to genetic colocalization analysis with cardiovascular-related associations identified by genome-wide association studies (GWAS), finding colocalized signals at 66 human disease loci corresponding to 30 genes and 39 variants. These genetically regulated splicing differences provide plausible mechanisms explaining some of the genetic risk for cardiovascular-related diseases. Among the top signals are novel isoforms of Endothelial Protein C Receptor (PROCR) and Distal Membrane Arm Assembly Component 2 (DMAC2), whose splicing patterns colocalize with risk for coronary artery disease (CAD). This study demonstrates the prevalence of inducible alternative promoters and supports that ECs express numerous novel transcripts regulated by genetics and inflammation that are consistent with driving individual risk for cardiovascular disease. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=66 SRC="FIGDIR/small/667484v1_ufig1.gif" ALT="Figure 1"> View larger version (18K): org.highwire.dtl.DTLVardef@1a702c6org.highwire.dtl.DTLVardef@1a71602org.highwire.dtl.DTLVardef@961cbaorg.highwire.dtl.DTLVardef@13802eb_HPS_FORMAT_FIGEXP M_FIG C_FIG