title: 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.
authors: Department of Molecular Cardiology, Cleveland Clinic, Cleveland, OH, USA; Regulatory Biology Graduate Program, Cleveland State University, Cleveland, OH, USA.
authors: Jacqueline R
authors: JR
authors: Kemp
authors: Department of Molecular Cardiology, Cleveland Clinic, Cleveland, OH, USA.
authors: Hamiyet
authors: H
authors: Unal
authors: Department of Molecular Cardiology, Cleveland Clinic, Cleveland, OH, USA.
authors: Russell
authors: R
authors: Desnoyer
authors: Department of Molecular Cardiology, Cleveland Clinic, Cleveland, OH, USA.
authors: Hong
authors: H
authors: Yue
authors: Department of Molecular Cardiology, Cleveland Clinic, Cleveland, OH, USA.
authors: Anushree
authors: A
authors: Bhatnagar
authors: Department of Molecular Cardiology, Cleveland Clinic, Cleveland, OH, USA; Regulatory Biology Graduate Program, Cleveland State University, Cleveland, OH, USA.
authors: Sadashiva S
authors: SS
authors: Karnik
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D001483:Base Sequence; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D016678:Genome / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism; D009928:Organ Specificity / Q000187:drug effects / Q000235:genetics; D051381:Rats; D044140:Receptor, Angiotensin, Type 1 / Q000378:metabolism; D012084:Renin-Angiotensin System / Q000187:drug effects* / Q000235:genetics*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C548912:MIRN483 microRNA, human; D035683:MicroRNAs; C548913:Mirn483 microRNA, mouse; D044140:Receptor, Angiotensin, Type 1; D000804:Angiotensin II
keywords: AT(1) receptor; Angiotensin II; Genome-wide expression; Renin–angiotensin system; Vascular smooth muscle; miRNA