Contribution of FBLN5 to Unstable Plaques in Carotid Atherosclerosis via mir128 and mir532–3p Based on Bioinformatics Prediction and Validation

Abstract

FBLN5, a member of the short fibulins in the fibulin family of extracellular matrix/matricellular proteins, is involved in interactions with components of the basement membrane and extracellular matrix proteins. It plays key roles in endothelial tissues in many vascular diseases. In this study, the relationship between FBLN5 and carotid atherosclerotic plaque stability as well as the regulatory roles of miRNAs were evaluated. Differential gene expression analyses and weighted gene co-expression network analysis (WGCNA) based on the GSE163154 dataset (including 16 samples without intraplaque hemorrhage and 27 samples with intraplaque hemorrhage) in GEO revealed that FBLN5 is related to plaque stability and is the most significantly differentially expressed gene. LASSO regression was used to evaluate genes obtained from the intersection of differentially expressed genes and clinically significant modules identified by WGCNA. A prediction model based on eight genes, including FBLN5, was constructed and showed an accuracy of 0.951 based on an ROC analysis. Low FBLN5 expression in plaque tissues was confirmed by immunohistochemistry and western blotting. GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analyses showed that FBLN5 acted mainly by the maintenance of the cellular matrix and reactive oxygen species production. miRNAs upstream of these eight predictive genes, including FBLN5, were identified and used to construct a network diagram. These results revealed that hsa-mir-128 and hsa-mir-532–3p were upstream regulatory factors of FBLN5, as verified by PCR assays of human plaque tissues demonstrating that both miRNAs were significantly up-regulated. Therefore, FBLN5 may play an important role in carotid atherosclerosis via hsa-mir-128 and hsa-mir-532–3p as well as become an essential target for treatment.

Identification of pivotal genes and regulatory networks associated with atherosclerotic carotid artery stenosis based on comprehensive bioinformatics analysis and machine learning

“…FBLN5 is a member of the fibronectin family and is essential for elastic fiber formation. It was discovered that FBLN5 may play an important role in carotid atherosclerosis via has-mir-128 and has-mir-532-3p ( Zheng et al, 2022 ). PPP1R12A, also known as MYPT1, is a key regulator of protein phosphatase 1C.…”

Differential Expression of Peripheral Circulating MicroRNA-146a Between Patients with Atherosclerotic Vulnerable Plaque and Stable Plaque

“…51) A bioinformatic study showed an association between miR-128 and plaque instability. 52) A study analyzed the role of miR-128 and miR-146a in macrophage-derived extracellular vesicles in the development of atherosclerosis and suggested that miR-146a might have an important role. 53) MiR-100 can suppress chronic vascular inflammation by stimulating autophagy in endothelial cells, inhibiting atherosclerosis progression.…”

LINC01089 blocks malignant progression of thyroid cancer by binding miR-27b-3p to enhance the FBLN5 protein level

“…Fibrin enhances cell-to-cell adhesion in the cellular microenvironment and also affects cell behavior [ 18 ]. Fibulin-5 (FBLN5), a fibrin, has been shown to enhance endothelial cell adhesion [ 19 ]. FBLN5 reduced the invasion, mammospheres formation, and proliferation capacity of cancer cells via silencing β-catenin phosphorylation in breast cancer and NSCLC, acting as a cancer suppressor gene [ 20 – 22 ].…”