iTRAQ Proteomics Identified the Potential Biomarkers of Coronary Artery Lesion in Kawasaki Disease and In Vitro Studies Demonstrated That S100A4 Treatment Made HCAECs More Susceptible to Neutrophil Infiltration

Abstract

Coronary artery lesions (CAL) are a major complication of Kawasaki disease (KD). The early prediction of CAL enables the medical personnel to apply adequate medical intervention. We collected the serum samples from the KD patients with CAL (n = 32) and those without CAL (n = 31), followed by a global screening with isobaric tagging for relative and absolute quantification (iTRAQ) technology and specific validation with an enzyme-linked immunosorbent assay (ELISA). iTRAQ identified 846 proteins in total in the serum samples, and four candidate proteins related to CAL were selected for ELISA validation as follows: Protein S100-A4 (S100A4), Catalase (CAT), Folate receptor gamma (FOLR3), and Galectin 10 (CLC). ELISA validation showed that the S100A4 level was significantly higher in KD patients with CAL than in those without CAL (225.2 ± 209.5 vs. 143.3 ± 83 pg/mL, p < 0.05). In addition, KD patients with CAL had a significantly lower CAT level than those without CAL (1.6 ± 1.5 vs. 2.7 ± 2.3 ng/mL, p < 0.05). Next, we found that S100A4 treatment on human coronary artery endothelial cells (HCAECs) reduced the abundance of cell junction proteins, which promoted the migration of HCAECs. Further assays also demonstrated that S100A4 treatment enhanced the permeability of the endothelial layer. These results concluded that S100A4 treatment resulted in an incompact endothelial layer and made HCAECs more susceptible to in vitro neutrophil infiltration. In addition, both upregulated S100A4 and downregulated CAT increased the risk of CAL in KD. Further in vitro study implied that S100A4 could be a potential therapeutic target for CAL in KD.

Cathelicidin (LL‐37) causes expression of inflammatory factors in coronary artery endothelial cells of Kawasaki disease by activating TLR4–NF‐κB–NLRP3 signaling

“…HCAECs are an important component of coronary arteries, and apoptosis of HCAECs is an essential factor leading to endothelial dysfunction in coronary arteries. Abnormally activated T cells and monocyte macrophages release a large number of cytokines and inflammatory mediators, which lead to endothelial cell dysfunction, impaired vascular function, lipid deposition, and proliferation of smooth muscle cells, ultimately leading to coronary artery lesions 25,26 . To further investigate the role of LL‐37, HCAECs were stimulated with human recombinant LL‐37, and as anticipated, LL‐37 significantly promoted apoptosis in HCAECs and significantly increased the expression levels of TLR4 and NLRP3.…”

“…Abnormally activated T cells and monocyte macrophages release a large number of cytokines and inflammatory mediators, which lead to endothelial cell dysfunction, impaired vascular function, lipid deposition, and proliferation of smooth muscle cells, ultimately leading to coronary artery lesions. 25 , 26 To further investigate the role of LL‐37, HCAECs were stimulated with human recombinant LL‐37, and as anticipated, LL‐37 significantly promoted apoptosis in HCAECs and significantly increased the expression levels of TLR4 and NLRP3. Previous studies reported that LL‐37 act on endothelial cells and contribute to the production of MCP‐1, ICAM‐1, affect the expression of cytokines IL‐1β, IL‐6, TNF‐a, and induce the production of IL‐17.…”

Effects of Adding Extracorporeal Shockwave Therapy (ESWT) to Platelet-Rich Plasma (PRP) among Patients with Rotator Cuff Partial Tear: A Prospective Randomized Comparative Study

“…By applying identification parameters of a false determinate rate <0.01 for protein and peptide identification and requiring protein identification with at least one unique peptide, 688 proteins were assessed and quantified. Abundance quantification of these proteins was further analyzed by using Partek [25]. With parameters set at p-value < 0.05 and variation >1.25, 15 proteins exhibited differential abundances between the two groups.…”

“…Isobaric tag for relative and absolute quantitation (iTRAQ) constitutes a mass spectrometrybased proteomic quantification technique that employs the derivatization of primary amino groups within intact proteins, along with isobaric tags assigned to distinct peptide fragments [20][21][22][23][24][25]. The iTRAQ methodology affords the opportunity to simultaneously screen up to eight samples for differentially expressed proteins with exceptional sensitivity and specificity [26].…”