RNA-sequencing analysis of differential gene expression associated with arterial stiffness

Vascular. 2020 Oct;28(5):655-663. doi: 10.1177/1708538120922650. Epub 2020 May 6.

Abstract

Objectives: Arterial stiffness is recognized as an important predictor of cardiovascular disease morbidity and mortality, independent of traditional cardiovascular disease risk factors. Given that arterial tissue is not easily accessible, most gene expression studies on arterial stiffness have been conducted on animals or on patients who have undergone by-pass surgeries. In order to obtain a deeper understanding of early changes of arterial stiffness, this study compared transcriptome profiles between healthy adults with higher and lower arterial stiffness.

Methods: The sample included 20 healthy female adults without cardiovascular disease. Arterial stiffness was measured by carotid-femoral pulse wave velocity, the "gold-standard" measure of central arterial stiffness. Peripheral blood samples collected to PAXgene™ RNA tubes were used for RNA sequencing (RNA-seq). The potential confounding effects of age, body mass index, and mean arterial pressure were controlled for in RNA-seq analysis. To validate RNA-seq results, quantitative real-time PCR (qRT-PCR) was performed for six selected genes.

Results: The findings demonstrated that genes including CAPN9, IL32, ERAP2, RAB6B, MYBPH, and miRNA626 were down-regulated, and that MOCS1 gene was up-regulated among the people with higher arterial stiffness. Real-time PCR showed that the changes of CAPN9, IL32, ERAP2, and RAB6B were in concordance with RNA-seq data, and confirmed the validity of the gene expression profiles obtained by RNA-seq analysis.

Conclusions: Previous studies have suggested the potential roles of CAPN9, IL32, and ERAP2 in structural changes of the arterial wall through up-regulation of metalloproteinases. However, the current study showed that CAPN9, IL32, and ERAP2 were down-regulated in the individuals with higher arterial stiffness, compared with those with lower arterial stiffness. The unexpected directions of expression of these genes may indicate an effort to maintain vascular homeostasis during increased arterial stiffness among healthy individuals. Further studies are guaranteed to investigate the roles of CAPN9, IL32, and ERAP2 in regulating arterial stiffness in people with and without cardiovascular disease.

Keywords: RNA-seq; arterial stiffness; pulse wave velocity; transcriptome.

MeSH terms

  • Adolescent
  • Adult
  • Aminopeptidases / genetics*
  • Arterial Pressure
  • Calpain / genetics*
  • Carotid-Femoral Pulse Wave Velocity
  • Down-Regulation
  • Female
  • Gene Expression Profiling*
  • Gene Regulatory Networks
  • Humans
  • Interleukins / genetics*
  • Middle Aged
  • RNA-Seq*
  • Transcriptome*
  • Vascular Stiffness / genetics*
  • Young Adult

Substances

  • IL32 protein, human
  • Interleukins
  • Aminopeptidases
  • ERAP2 protein, human
  • CAPN9 protein, human
  • Calpain