title: MicroRNA-1 regulates smooth muscle cell differentiation by repressing Kruppel-like factor 4.
abstract: The role of microRNA-1 (miR-1) has been studied in cardiac and skeletal muscle differentiation. However, it remains unexplored in vascular smooth muscle cells (SMCs) differentiation. The aim of this study was to uncover novel targets of and shed light on the function of miR-1 in the context of embryonic stem cell (ESC) differentiation of SMCs in vitro. miR-1 expression is steadily increased during differentiation of mouse ESC to SMCs. Loss-of-function approaches using miR-1 inhibitors uncovered that miR-1 is required for SMC lineage differentiation in ESC-derived SMC cultures, as evidenced by downregulation of SMC-specific markers and decrease of derived SMC population. In addition, bioinformatics analysis unveiled a miR-1 binding site on the Kruppel-like factor 4 (KLF4) 3' untranslated region (3'UTR), in a region that is highly conserved across species. Consistently, miR-1 mimic reduced KLF4 3'UTR luciferase activity, which can be rescued by mutating the miR-1 binding site on the KLF4 3'UTR in the reporter construct. Additionally, repression of the miR-1 expression by miR-1 inhibitor can reverse KLF4 downregulation during ESC-SMC differentiation, which subsequently inhibits SMC differentiation. We conclude that miR-1 plays a critical role in the determination of SMC fate during retinoid acid-induced ESC/SMC differentiation, which may indicate that miR-1 has a role to promote SMC differentiation.
authors.affiliation: Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, Michigan 48109, USA.
authors.forename: Changqing
authors.initials: C
authors.lastname: Xie
authors.forename: Huarong
authors.initials: H
authors.lastname: Huang
authors.forename: Xuan
authors.initials: X
authors.lastname: Sun
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Guo
authors.forename: Milton
authors.initials: M
authors.lastname: Hamblin
authors.forename: Raquel P
authors.initials: RP
authors.lastname: Ritchie
authors.forename: Minerva T
authors.initials: MT
authors.lastname: Garcia-Barrio
authors.forename: Jifeng
authors.initials: J
authors.lastname: Zhang
authors.forename: Y Eugene
authors.initials: YE
authors.lastname: Chen
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D001665:Binding Sites; D002454:Cell Differentiation; D019295:Computational Biology; D015536:Down-Regulation; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism; D034622:RNA Interference; D015854:Up-Regulation
chemical_list: D020413:3' Untranslated Regions; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D035683:MicroRNAs; C517731:Mirn1 microRNA, mouse
references.citation: Nat Genet. 2005 May;37(5):495-500
references.pmid: 15806104
references.citation: J Biol Chem. 2001 Aug 10;276(32):30423-8
references.pmid: 11390382
references.citation: Curr Biol. 2002 Apr 30;12(9):735-9
references.pmid: 12007417
references.citation: Arterioscler Thromb Vasc Biol. 2009 Jan;29(1):99-106
references.pmid: 18927467
references.citation: Genes Dev. 2009 Sep 15;23(18):2166-78
references.pmid: 19720868
references.citation: Science. 2001 Oct 26;294(5543):853-8
references.pmid: 11679670
references.citation: Circ Res. 2007 Oct 12;101(8):792-801
references.pmid: 17704209
references.citation: J Biol Chem. 2006 Jun 23;281(25):16951-16961
references.pmid: 16632465
references.citation: Arterioscler Thromb Vasc Biol. 2007 Dec;27(12):e311-2
references.pmid: 18029907
references.citation: Cell. 2003 Dec 26;115(7):787-98
references.pmid: 14697198
references.citation: Circ Res. 2008 Oct 24;103(9):919-28
references.pmid: 18948630
references.citation: Nat Genet. 2003 Nov;35(3):215-7
references.pmid: 14528307
references.citation: Arterioscler Thromb Vasc Biol. 2005 Jun;25(6):1135-41
references.pmid: 15817882
references.citation: Curr Opin Cell Biol. 2009 Jun;21(3):461-9
references.pmid: 19278845
references.citation: Mol Endocrinol. 2009 May;23(5):724-33
references.pmid: 19196830
references.citation: Stem Cells Dev. 2009 Jun;18(5):741-8
references.pmid: 18795840
references.citation: J Biol Chem. 2009 Feb 6;284(6):3728-38
references.pmid: 19088079
references.citation: J Biol Chem. 2010 Mar 26;285(13):9383-9389
references.pmid: 20118242
references.citation: Am J Physiol Cell Physiol. 2008 Nov;295(5):C1175-82
references.pmid: 18768922
references.citation: Circ Res. 2008 Jun 20;102(12):1548-57
references.pmid: 18483411
references.citation: Circ Res. 2007 Jun 8;100(11):1579-88
references.pmid: 17478730
references.citation: Cell. 2006 Dec 15;127(6):1193-207
references.pmid: 17174894
references.citation: Circ Res. 2009 Jul 17;105(2):158-66
references.pmid: 19542014
references.citation: Circ Res. 2009 Feb 27;104(4):476-87
references.pmid: 19150885
references.citation: J Biol Chem. 2003 Mar 28;278(13):11661-9
references.pmid: 12538588
references.citation: J Biol Chem. 2009 Mar 20;284(12):7903-13
references.pmid: 19158092
references.citation: Am J Physiol Heart Circ Physiol. 2009 Apr;296(4):H1027-37
references.pmid: 19168719
references.citation: Am J Physiol Cell Physiol. 2007 Jan;292(1):C59-69
references.pmid: 16956962
references.citation: Nature. 2009 Aug 6;460(7256):705-10
references.pmid: 19578358
references.citation: J Biol Chem. 2005 Mar 11;280(10):9719-27
references.pmid: 15623517
references.citation: Nature. 2008 Jul 3;454(7200):56-61
references.pmid: 18548003
references.citation: J Biol Chem. 2003 Nov 28;278(48):48004-11
references.pmid: 12970361
references.citation: Trends Biochem Sci. 2004 Sep;29(9):462-8
references.pmid: 15337119
references.citation: Curr Opin Chem Biol. 2003 Aug;7(4):516-23
references.pmid: 12941428