Atherosclerotic Conditions Promote the Packaging of Functional MicroRNA-92a-3p Into Endothelial Microvesicles

Abstract

Rationale: Microvesicle-incorporated microRNAs (miRs) are biomarkers and effectors of cardiovascular disease. Whether microvesicle-miR expression is regulated in coronary artery disease (CAD) or not is unknown. Objective: Here, we explore the expression of circulating microvesicle-miRs in patients with CAD and investigate the role of microvesicle-miR in endothelial cells. Methods and Results: Circulating microvesicles were isolated from patients’ plasma by using ultracentrifugation. Electron microscopy was used to determine the size of the microvesicles. A Taqman miR array revealed certain microvesicle-miRs are significantly regulated in patients with stable CAD compared with patients with ACS. To validate the miR array results, 180 patients with angiographically excluded CAD (n=41), stable CAD (n=77), and acute coronary syndrome (n=62) were prospectively studied. Nine miRs involved in regulation of vascular performance—miR-126-3p, miR-222-3p, miR-let-7d-5p, miR-21-5p, miR-26a-5p, miR-92a-3p, miR-139-5p, miR-30b-5p, and miR-199a-5p—were quantified in circulating microvesicles by real-time polymerase chain reaction (PCR). Among these, miR-92a-3p was significantly increased in patients with CAD compared with non-CAD patients. Microvesicle-sorting experiments showed endothelial cells (ECs) were the major cell source for microvesicles containing miR-92a-3p. In vitro oxLDL (oxidized low-density lipoprotein) and IL-6 (interleukin-6) stimulation increased miR-92a-3p expression in parent ECs and upregulated the expression level of endothelial microvesicle (EMV)-incorporated miR-92a-3p. Labeling of miR-92a-3p and EMVs demonstrated that functional miR-92a-3p was transported into recipient ECs, which accelerated cell migration and proliferation. Knockdown of miR-92a-3p in EMVs abrogated EMV-mediated effects on EC migration, proliferation, and blocked vascular network formation in a matrigel plug. Polymerase chain reaction–based gene profiling showed that the expression of THBS1 (thrombospondin 1) protein—a target of miR-92a-3p and an inhibitor of angiogenesis—was significantly reduced in ECs by EMVs. Knockdown of miR-92a-3p in EMVs abrogated EMV-mediated inhibition of the THBS1 gene and protein expression. Conclusions: Atherosclerotic conditions promote the packaging of endothelial miR-92a-3p into EMVs. EMV-mediated transfer of functional miR-92a-3p regulates angiogenesis in recipient ECs by a THBS1-dependent mechanism.

Sorting Mechanisms for MicroRNAs into Extracellular Vesicles and Their Associated Diseases

“…Atherosclerosis is widely known to be the driving force behind coronary artery disease and high levels of oxidized LDL are a significant risk factor for atherosclerosis development and progression [82]. Recently, it has been demonstrated that oxidized LDL (oxLDL) has the ability to selectively shuttle miRNAs into MVs [83]. Specifically, miR-92a-3p, miR-222-3p, and miR-26a-5p were selectively sorted into MVs through an oxLDL-dependent mechanism.…”

Prognostic value of Ki-67 in patients with hypertension and prostate cancer: A real-world study in a Chinese population

“…Hypertension and ACS are pathological conditions that damage the endothelium, triggering cell proliferation, vascular remodeling, and other malignant biological behaviors. [41,42] A previous study considered that the expression of Ki-67 was lower in smokers than non-smokers, [43] but the mechanisms underlying this are still unclear. A meta-analysis reported that low Ki-67 is associated with a lower risk of developing diabetes mellitus, [33] but our study did not find an association between Ki-67 and diabetes mellitus.…”

MiR-133a-3p transferred by circulating microvesicles derived from myocardial ischemic preconditioning protects cardiomyocytes against hypoxia/reoxygenation injury

“…As a kind of paracrine factor, MVs allow the transfer of cytoplasmic components such as proteins, mRNAs and miRNAs from one cell to another [8]. However, circulating MVs vary in their components, which varies with the different parent cell types or different stimuli, thus playing different biological roles in occurrence and development of diseases [9,12].…”

“…MVs are submicron membrane vesicles with the size range of 100 ~ 1000 nm in diameter shed from almost all eukaryotic cells during activation or apoptosis [7]. Multiple experiments have shown that MVs derived from various cell types could play intercellular communication role in diagnostic and therapeutic effects of cardiovascular diseases, mainly by delivering proteins, cytokines and gene messages such as mRNAs and microRNAs (miRNAs) to target cells [8]. Among the components of MVs, miRNAs have been shown to play important roles that regulate the pathophysiological consequences of myocardial I/R injury [9], which can be released from various cells by MVs that protect them from degradation and guarantee their stability and circulation through the blood stream [10], suggesting miRNA in MVs could be a potent therapeutic target and tool for protecting myocardium against I/R injury.…”

“…Instead, transfection with the miR-133a-3p inhibitor could aggravate the ERS-induced apoptosis through increasing the expression of GRP78, CHOP and caspase 12. Hence, the suppression of ERS-induced apoptosis may be involved in the protective effect of miR-133a-3p on H/R injured H9c2 cells, consistent with IPC-MVs.Recent studies have suggested that MVs can transfer functional miRNAs between cells by variousroutes, including endocytotic uptaking[8]. Additionally, cardiomyocytes can release and take up MVs both in vivo and in vitro[13].…”