Systems Genetics in Human Endothelial Cells Identifies Non-coding Variants Modifying Enhancers, Expression, and Complex Disease Traits

Am J Hum Genet. 2020 Jun 4;106(6):748-763. doi: 10.1016/j.ajhg.2020.04.008. Epub 2020 May 21.


The identification of causal variants and mechanisms underlying complex disease traits in humans is important for the progress of human disease genetics; this requires finding strategies to detect functional regulatory variants in disease-relevant cell types. To achieve this, we collected genetic and transcriptomic data from the aortic endothelial cells of up to 157 donors and four epigenomic phenotypes in up to 44 human donors representing individuals of both sexes and three major ancestries. We found thousands of expression quantitative trait loci (eQTLs) at all ranges of effect sizes not detected by the Gene-Tissue Expression Project (GTEx) in human tissues, showing that novel biological relationships unique to endothelial cells (ECs) are enriched in this dataset. Epigenetic profiling enabled discovery of over 3,000 regulatory elements whose activity is modulated by genetic variants that most frequently mutated ETS, AP-1, and NF-kB binding motifs, implicating these motifs as governors of EC regulation. Using CRISPR interference (CRISPRi), allele-specific reporter assays, and chromatin conformation capture, we validated candidate enhancer variants located up to 750 kb from their target genes, VEGFC, FGD6, and KIF26B. Regulatory SNPs identified were enriched in coronary artery disease (CAD) loci, and this result has specific implications for PECAM-1, FES, and AXL. We also found significant roles for EC regulatory variants in modifying the traits pulse pressure, blood protein levels, and monocyte count. Lastly, we present two unlinked SNPs in the promoter of MFAP2 that exhibit pleiotropic effects on human disease traits. Together, this supports the possibility that genetic predisposition for complex disease is manifested through the endothelium.

Keywords: association mapping; complex disease; endothelial cells; epigenetics; genomics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Disease / genetics*
  • Endothelial Cells / metabolism*
  • Enhancer Elements, Genetic / genetics*
  • Epigenesis, Genetic / genetics
  • Female
  • Gene Expression Regulation / genetics*
  • Genetic Variation / genetics*
  • Humans
  • Kinesin / genetics
  • Male
  • Mutation
  • NF-kappa B / metabolism
  • Polymorphism, Single Nucleotide / genetics
  • Proto-Oncogene Protein c-ets-1 / metabolism
  • Quantitative Trait Loci / genetics
  • Transcription Factor AP-1 / metabolism
  • Transcriptional Regulator ERG / metabolism
  • Vascular Endothelial Growth Factor C / genetics


  • ERG protein, human
  • NF-kappa B
  • Proto-Oncogene Protein c-ets-1
  • Transcription Factor AP-1
  • Transcriptional Regulator ERG
  • VEGFC protein, human
  • Vascular Endothelial Growth Factor C
  • KIF26B protein, human
  • Kinesin