title: Small RNA sequencing reveals microRNAs that modulate angiotensin II effects in vascular smooth muscle cells.
abstract: Angiotensin II (Ang II)-mediated vascular smooth muscle cell dysfunction plays a critical role in cardiovascular diseases. However, the role of microRNAs (miRNAs) in this process is unclear. We used small RNA deep sequencing to profile Ang II-regulated miRNAs in rat vascular smooth muscle cells (VSMC) and evaluated their role in VSMC dysfunction. Sequencing results revealed several Ang II-responsive miRNAs, and bioinformatics analysis showed that their predicted targets can modulate biological processes relevant to cardiovascular diseases. Further studies with the most highly induced miR-132 and miR-212 cluster (miR-132/212) showed time- and dose-dependent up-regulation of miR-132/212 by Ang II through the Ang II Type 1 receptor. We identified phosphatase and tensin homolog (PTEN) as a novel target of miR-132 and demonstrated that miR-132 induces monocyte chemoattractant protein-1 at least in part via PTEN repression in rat VSMC. Moreover, miR-132 overexpression enhanced cyclic AMP-response element-binding protein (CREB) phosphorylation via RASA1 (p120 Ras GTPase-activating protein 1) down-regulation, whereas miR-132 inhibition attenuated Ang II-induced CREB activation. Furthermore, miR-132 up-regulation by Ang II required CREB activation, demonstrating a positive feedback loop. Notably, aortas from Ang II-infused mice displayed similar up-regulation of miR-132/212 and monocyte chemoattractant protein-1, supporting in vivo relevance. In addition, microarray analysis and reverse transcriptase-quantitative PCR validation revealed additional novel miR-132 targets among Ang II-down-regulated genes implicated in cell cycle, motility, and cardiovascular functions. These results suggest that miR132/212 can serve as a novel cellular node to fine-tune and amplify Ang II actions in VSMC.
authors.affiliation: Irell and Manella Graduate School of Biological Sciences, Beckman Research Institute of City of Hope, Duarte, California 91010, USA.
authors.forename: Wen
authors.initials: W
authors.lastname: Jin
authors.forename: Marpadga A
authors.initials: MA
authors.lastname: Reddy
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Chen
authors.forename: Sumanth
authors.initials: S
authors.lastname: Putta
authors.forename: Linda
authors.initials: L
authors.lastname: Lanting
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Kato
authors.forename: Jung Tak
authors.initials: JT
authors.lastname: Park
authors.forename: Manasa
authors.initials: M
authors.lastname: Chandra
authors.forename: Charles
authors.initials: C
authors.lastname: Wang
authors.forename: Rajendra K
authors.initials: RK
authors.lastname: Tangirala
authors.forename: Rama
authors.initials: R
authors.lastname: Natarajan
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000378:metabolism; D051059:PTEN Phosphohydrolase / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D020413:3' Untranslated Regions; C491766:Ccl2 protein, rat; D018932:Chemokine CCL2; D017362:Cyclic AMP Response Element-Binding Protein; D018076:DNA, Complementary; C548821:MIRN132 microRNA, rat; C552427:MIRN212 microRNA, rat; D035683:MicroRNAs; D000804:Angiotensin II; D051059:PTEN Phosphohydrolase