Increased expression of miR-224-5p in circulating extracellular vesicles of patients with reduced coronary flow reserve

Abstract

Background Endothelial and microvascular dysfunction are pivotal causes of major adverse cardiac events predicted by coronary flow reserve (CFR). Extracellular Vesicles (EVs) have been studied extensively in the pathophysiology of coronary artery disease. However, little is known on the impact of the non-coding RNA content of EVs with respect to CFR. Methods We carried out a study among 120 patients divided by high-CFR and low-CFR to profile the miRNA content of circulating EVs. Results A multiplex array profiling on circulating EVs revealed mir-224-5p (p-value ≤ 0.000001) as the most differentially expressed miRNA in the Low-CFR group and showed a significantly independent relationship to CFR. Literature survey indicated the origin of the miR from liver cells and not of platelet, leukocyte, smooth muscle or endothelial (EC) origin. A q-PCR panel of the conventional cell type-EVs along with hepatic EVs showed that EVs from liver cells showed higher expression of the miR-224-5p. FACS analysis demonstrated the presence of liver-specific (ASGPR-1+/CD14−) EVs in the plasma of our cohort with the presence of Vanin-1 required to enter the EC barrier. Hepatic EVs with and without the miR-224-5p were introduced to ECs in-vitro, but with no difference in effect on ICAM-1 or eNOS expression. However, hepatic EVs elevated endothelial ICAM-1 levels per se independent of the miR-224-5p. Conclusion This indicated a role of hepatic EVs identified by the miR-224-5p in endothelial dysfunction in patients with Low CFR.

miRNA Dysregulation in Cardiovascular Diseases: Current Opinion and Future Perspectives

“…Eyyupkoca et al [ 69 ] described that eight miRNAs (miR-23b-3p, miR-26b-5p, miR-199a-5p, miR-301a-3p, miR-374a-5p, miR-423-5p, miR-483-5p, and miR-652-3p) were associated with adverse left ventricular remodeling (ALVR), although only three, miR-301a-3p, miR-374a-5p, and miR-423-5p, in a significative manner. James et al [ 71 ] performed a multiplex array profiling, demonstrating that miR-224-5p is more expressed in circulating vesicles of patients with reduced coronary flow reserve. Barbalata et al [ 33 ] concluded that plasma levels of miR-142, miR-223, and miR-155 were higher in peripheral artery disease (PAD) patients with cardiovascular events (CVEs) compared with those without CVEs, while miR-92a is lower.…”

Liquid Biopsy in the Clinical Management of Cancers

“…EV Proteomic and RNA Profiling The integration of proteomic and RNA profiling in EV research holds great potential for advancing precision medicine and improving disease diagnosis and monitoring. The development of EV proteomic profiling has been promising for the detection and monitoring of cancer, emphasizing the importance of understanding the protein cargo of EVs for clinical applications [19,148]. Through the use of high-throughput sequencing and NGS, comprehensive analyses of EV RNA content have been carried out, which have shed light on the multitude of RNA species present in EVs as well as the functional implications of their presence [130].…”

The Roles of Extracellular Vesicles in the Progression of Renal Cell Carcinoma and Their Potential for Future Clinical Application

“…However, elevated EV levels of miR-224 are not specific to RCC. miR-224 is demonstrated to function as an oncogenic miRNA in several types of cancer, including hepatocellular carcinoma, non-small-cell lung cancer, and colorectal cancer [ 122 , 123 ]. Du et al found that the miR-let-7i-5p, miR-26a-1-3p, and miR-615-3p levels in plasma EVs are associated with the overall survival (OS) of mRCC patients.…”