Antagomir-mediated silencing of endothelial cell specific microRNA-126 impairs ischemia-induced angiogenesis.
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.
van Solingen
Coen
C
Department of Nephrology and the Einthoven Laboratory for Experimental Vascular Medicine, LUMC, Leiden, The Netherlands.
Seghers
Leonard
L
Bijkerk
Roel
R
Duijs
Jacques M G J
JM
Roeten
Marko K
MK
van Oeveren-Rietdijk
Annemarie M
AM
Baelde
Hans J
HJ
Monge
Matthieu
M
Vos
Joost B
JB
de Boer
Hetty C
HC
Quax
Paul H A
PH
Rabelink
Ton J
TJ
van Zonneveld
Anton Jan
AJ
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
C545201:MIRN126 microRNA, mouse; D035683:MicroRNAs; D009843:Oligoribonucleotides