Angiotensin II-regulated microRNA 483-3p directly targets multiple components of the renin–angiotensin system

Abstract

Improper regulation of signaling in vascular smooth muscle cells (VSMCs) by angiotensin II (AngII) can lead to hypertension, vascular hypertrophy and atherosclerosis. The extent to which the homeostatic levels of the components of signaling networks are regulated through microRNAs (miRNA) modulated by AngII type 1 receptor (AT1R) in VSMCs is not fully understood. Whether AT1R blockers used to treat vascular disorders modulate expression of miRNAs is also not known. To report differential miRNA expression following AT1R activation by AngII, we performed microarray analysis in 23 biological and technical replicates derived from humans, rats and mice. Profiling data revealed a robust regulation of miRNA expression by AngII through AT1R, but not the AngII type 2 receptor (AT2R). The AT1R-specific blockers, losartan and candesartan antagonized >90% of AT1R-regulated miRNAs and AngII-activated AT2R did not modulate their expression. We discovered VSMC-specific modulation of 22 miRNAs by AngII, and validated AT1R-mediated regulation of 17 of those miRNAs by real-time polymerase chain reaction analysis. We selected miR-483-3p as a novel representative candidate for further study because mRNAs of multiple components of the renin angiotensin system (RAS) were predicted to contain the target sequence for this miRNA. MiR-483-3p inhibited the expression of luciferase reporters bearing 3′-UTRs of four different RAS genes and the inhibition was reversed by antagomir-483-3p. The AT1R-regulated expression levels of angiotensinogen and Angiotensin Converting Enzyme 1 (ACE-1) proteins in VSMCs are modulated specifically by miR-483-3p. Our study demonstrates that the AT1R-regulated miRNA expression fingerprint is conserved in VSMCs of humans and rodents. Furthermore, we identify the AT1R-regulated miR-483-3p as a potential negative regulator of steady-state levels of RAS components in VSMCs. Thus, miRNA-regulation by AngII to affect cellular signaling is a novel aspect of RAS biology, which may lead to discovery of potential candidate prognostic markers and therapeutic targets.

International Union of Basic and Clinical Pharmacology. XCIX. Angiotensin Receptors: Interpreters of Pathophysiological Angiotensinergic Stimuli

“…In human coronary artery endothelial cells, lentiviral delivery of the AT 2 receptor changes expression of a large number of genes without AT 2 receptor ligands, and many fewer genes were differentially expressed when the AT 2 receptorspecific ligand CGP42112 was added (Falcon et al, 2005). Kemp et al (2014b) found that AT 2 receptor expression antagonized regulation of microRNAs by AT 1 receptor and AngII stimulation of AT 2 receptor affected expression of only a few microRNAs, whereas the same treatment caused a robust response from AT 1 receptor. These findings suggest that altered expression of AT 2 receptor itself is a stimulus for function and that many cellular effects of AT 2 receptor expression are not contingent on ligand interaction with this receptor.…”

Hydroxytyrosol supplementation modulates the expression of miRNAs in rodents and in humans

“…Although potential modulation of selected miRNAs by HT was not fully consistent in all models studied, miR-483-3p stood out as a robust candidate. miR-483-3p has been associated with hypertension [33], apoptosis [34] and proliferation [35]. miR-483-3p has been also associated with the capacity of adipocytes to store lipids and dysregulation of its expression could directly inhibit adipose tissue expandability and lipid storage, which in turn affect other tissues by stimulating ectopic triglyceride storage and lipotoxicity [36].…”

miR-483-3p regulates hyperglycaemia-induced cardiomyocyte apoptosis in transgenic mice

“…The expression of miR-483 has been evaluated in diseases such as liver cancer and cartilage cancer, and we have investigated its function in ischemic heart disease [14,23]. In vascular smooth muscles, abundantly found in blood vessels, angiotensin II-regulated miR-483-3p negatively affects homeostasis between the components of the renin-angiotensin system [24]. It has also been suggested that circulating miR-483-3p can serve as a potential biomarker in predicting and monitoring patient response to left ventricular assist device therapy [25].…”