title: Antagomir-mediated silencing of endothelial cell specific microRNA-126 impairs ischemia-induced angiogenesis.
abstract: MicroRNAs are negative regulators of gene expression that play a key role in cell-type specific differentiation and modulation of cell function and have been proposed to be involved in neovascularization. Previously, using an extensive cloning and sequencing approach, we identified miR-126 to be specifically and highly expressed in human endothelial cells (EC). Here, we demonstrate EC-specific expression of miR-126 in capillaries and the larger vessels in vivo. We therefore explored the potential role of miR-126 in arteriogenesis and angiogenesis. Using miR-reporter constructs, we show that miR-126 is functionally active in EC in vitro and that it could be specifically repressed using antagomirs specifically targeting miR-126. To study the consequences of miR-126 silencing on vascular regeneration, mice were injected with a single dose of antagomir-126 or a control 'scramblemir' and exposed to ischemia of the left hindlimb by ligation of the femoral artery. Although miR-126 was effectively silenced in mice treated with a single, high dose (HD) of antagomir-126, laser Doppler perfusion imaging did not show effects on blood flow recovery. In contrast, quantification of the capillary density in the gastrocnemius muscle revealed that mice treated with a HD of antagomir-126 had a markedly reduced angiogenic response. Aortic explant cultures of the mice confirmed the role of miR-126 in angiogenesis. Our data demonstrate a facilitary function for miR-126 in ischemia-induced angiogenesis and show the efficacy and specificity of antagomir-induced silencing of EC-specific microRNAs in vivo.
authors.affiliation: Department of Nephrology and the Einthoven Laboratory for Experimental Vascular Medicine, LUMC, Leiden, The Netherlands.
authors.forename: Coen
authors.initials: C
authors.lastname: van Solingen
authors.forename: Leonard
authors.initials: L
authors.lastname: Seghers
authors.forename: Roel
authors.initials: R
authors.lastname: Bijkerk
authors.forename: Jacques M G J
authors.initials: JM
authors.lastname: Duijs
authors.forename: Marko K
authors.initials: MK
authors.lastname: Roeten
authors.forename: Annemarie M
authors.initials: AM
authors.lastname: van Oeveren-Rietdijk
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Baelde
authors.forename: Matthieu
authors.initials: M
authors.lastname: Monge
authors.forename: Joost B
authors.initials: JB
authors.lastname: Vos
authors.forename: Hetty C
authors.initials: HC
authors.lastname: de Boer
authors.forename: Paul H A
authors.initials: PH
authors.lastname: Quax
authors.forename: Ton J
authors.initials: TJ
authors.lastname: Rabelink
authors.forename: Anton Jan
authors.initials: AJ
authors.lastname: van Zonneveld
mesh_terms: D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000473:pathology; D001158:Arteries / Q000187:drug effects / Q000196:embryology; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D005786:Gene Expression Regulation / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects*; D006801:Humans; D007511:Ischemia / Q000473:pathology*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D009843:Oligoribonucleotides / Q000494:pharmacology*; D009928:Organ Specificity / Q000187:drug effects
chemical_list: C545201:MIRN126 microRNA, mouse; D035683:MicroRNAs; D009843:Oligoribonucleotides