miRNome Profiling in Bicuspid Aortic Valve-Associated Aortopathy by Next-Generation Sequencing

Int J Mol Sci. 2017 Nov 22;18(11):2498. doi: 10.3390/ijms18112498.

Abstract

The molecular mechanisms underlying thoracic aortic aneurysm (TAA) in patients with bicuspid aortic valve (BAV) are incompletely characterized. MicroRNAs (miRNAs) may play a major role in the different pathogenesis of aortopathy. We sought to employ next-generation sequencing to analyze the entire miRNome in TAA tissue from patients with BAV and tricuspid aortic valve (TAV). In the discovery stage, small RNA sequencing was performed using the Illumina MiSeq platform in 13 TAA tissue samples (seven patients with BAV and six with TAV). Gene ontology (GO) and KEGG pathway analysis were used to identify key pathways and biological functions. Validation analysis was performed by qRT-PCR in an independent cohort of 30 patients with BAV (26 males; 59.5 ± 12 years) and 30 patients with TAV (16 males; 68.5 ± 9.5 years). Bioinformatic analysis identified a total of 489 known mature miRNAs and five novel miRNAs. Compared to TAV samples, 12 known miRNAs were found to be differentially expressed in BAV, including two up-regulated and 10 down-regulated (FDR-adjusted p-value ≤ 0.05 and fold change ≥ 1.5). GO and KEGG pathway enrichment analysis (FDR-adjusted p-value < 0.05) identified different target genes and pathways linked to BAV and aneurysm formation, including Hippo signaling pathway, ErbB signaling, TGF-beta signaling and focal adhesion. Validation analysis of selected miRNAs confirmed the significant down-regulation of miR-424-3p (p = 0.01) and miR-3688-3p (p = 0.03) in BAV patients as compared to TAV patients. Our study provided the first in-depth screening of the whole miRNome in TAA specimens and identified specific dysregulated miRNAs in BAV patients.

Keywords: BAV; NGS; TAV; aortopathy; miRNome.

MeSH terms

  • Aortic Valve / abnormalities*
  • Aortic Valve / metabolism
  • Aortic Valve / pathology
  • Bicuspid Aortic Valve Disease
  • Computational Biology / methods
  • Female
  • Gene Expression Profiling
  • Gene Ontology
  • Heart Valve Diseases / genetics*
  • Heart Valve Diseases / metabolism
  • Heart Valve Diseases / pathology*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Male
  • MicroRNAs / genetics*
  • Molecular Sequence Annotation
  • Reproducibility of Results
  • Transcriptome*

Substances

  • MicroRNAs