Role of neutrophil extracellular trap and immune infiltration in atherosclerotic plaque instability: Novel insight from bioinformatics analysis and machine learning

Abstract

The instability of atherosclerotic plaques increases the risk of acute coronary syndrome. Neutrophil extracellular traps (NETs), mesh-like complexes consisting of extracellular DNA adorned with various protein substances, have been recently discovered to play an essential role in atherosclerotic plaque formation and development. This study aimed to investigate novel diagnostic biomarkers that can identify unstable plaques for early distinction and prevention of plaque erosion or disruption. Differential expression analysis was used to identify the differentially expressed NET-related genes, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed. We filtered the characteristic genes using machine learning and estimated diagnostic efficacy using receiver operating characteristic curves. Immune infiltration was detected using single-sample gene set enrichment analysis and the biological signaling pathways involved in characteristic genes utilizing gene set enrichment analysis were explored. Finally, miRNAs- and transcription factors-target genes networks were established. We identified 8 differentially expressed NET-related genes primarily involved in immune-related pathways. Four were identified as capable of distinguishing unstable plaques. More immune cells infiltrated unstable plaques than stable plaques, and these cells were predominantly positively related to characteristic genes. These 4 diagnostic genes are involved in immune responses and the modulation of smooth muscle contractility. Several miRNAs and transcription factors were predicted as upstream regulatory factors, providing further information on the identification and prevention of atherosclerotic plaques rupture. We identified several promising NET-related genes (AQP9, C5AR1, FPR3, and SIGLEC9) and immune cell subsets that may identify unstable atherosclerotic plaques at an early stage and prevent various complications of plaque disruption.