Candidate biomarker identification for blood stasis syndrome among coronary artery disease patients using the Olink proteomics platform.

Abstract

OBJECTIVE: To identify candidate biomarkers of blood stasis syndrome (BSS) associated with coronary artery disease (CAD) and explore the underlying inflammatory mechanisms. METHODS: Using the Olink Target 96 Inflammation panel, we identified plasma proteins in a group of 88 patients comprised of healthy controls (HCs), those with CAD and BSS (CAD-BSS), those with CAD without BSS (CAD-non-BSS), and those with BSS without CAD (non-CAD-BSS) (n = 22 in each group). Protein molecules that were specifically expressed in CAD or BSS were identified by differential expression analyses. Subsequently, potential protein biomarkers were identified using least absolute shrinkage and selection operator regression to enable CAD and BSS differentiation. The potential functional mechanisms of identified proteins were then determined by Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses. RESULTS: Patients with CAD had 31/92 upregulated and 4/92 downregulated proteins compared with those without. Chemokine (C-C motif) ligand 11 (CCL11), CUB domain-containing protein 1, hepatocyte growth factor, sirtuin 2 (SIRT2), eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), CCL25, and tumor necrosis factor (TNF) showed the strongest upregulation (all P <0.0001). Patients with BSS had 8/92 downregulated proteins, specifically CCL28, CCL11, cystatin D, STAM-binding protein, 4E-BP1, matrix metalloproteinase-10, SIRT2, and monocyte chemotactic protein 4, compared with those without (all P < 0.05). The CAD-BSS group had one interleukin-17 (IL-17) upregulated and 10/92 downregulated proteins compared with the CAD-non-BSS group. When compared with the non-CAD-BSS group, the CAD-BSS group had 8 upregulated proteins but only 2 downregulated proteins, namely interleukin-10 receptor subunit alpha (IL-10RA) and TNF-related activation-induced cytokine (both P < 0.05). Totally 10 proteins were identified as potential candidate biomarkers of BSS in CAD patients. After least absolute shrinkage and selection operator regression analysis, two proteins that distinguished between BSS and non-BSS individuals among CAD patients were identified (SIRT2 and 4E-BP1). These proteins are primarily associated with the mechanistic target of rapamycin signaling pathway, which regulates inflammation and oxidative stress. CONCLUSIONS: Results suggest that the inflammatory response and mechanistic target of rapamycin signaling pathway participate in CAD and BSS development, and that SIRT2 and 4E-BP1 are prospective protein biomarkers for patients with CAD and BSS.