Vascular endothelial cell-specific microRNA-15a inhibits angiogenesis in hindlimb ischemia.
The effects and potential mechanisms of the vascular endothelial cell (EC)-enriched microRNA-15a (miR-15a) on angiogenesis remain unclear. Here, we show a novel finding that EC-selective miR-15a transgenic overexpression leads to reduced blood vessel formation and local blood flow perfusion in mouse hindlimbs at 1-3 weeks after hindlimb ischemia. Mechanistically, gain- or loss-of-miR-15a function by lentiviral infection in ECs significantly reduces or increases tube formation, cell migration, and cell differentiation, respectively. By FGF2 and VEGF 3'-UTR luciferase reporter assays, Real-time PCR, and immunoassays, we further identified that the miR-15a directly targets FGF2 and VEGF to facilitate its anti-angiogenic effects. Our data suggest that the miR-15a in ECs can significantly suppress cell-autonomous angiogenesis through direct inhibition of endogenous endothelial FGF2 and VEGF activities. Pharmacological modulation of miR-15a function may provide a new therapeutic strategy to intervene against angiogenesis in a variety of pathological conditions.
Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, Michigan 48109, USA. kejie@umich.edu
Ke-Jie
KJ
Yin
Karl
K
Olsen
Milton
M
Hamblin
Jifeng
J
Zhang
Steven P
SP
Schwendeman
Y Eugene
YE
Chen
D020413:3' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D016222:Fibroblast Growth Factor 2 / Q000037:antagonists & inhibitors; D005455:Fluorescent Antibody Technique; D006614:Hindlimb / Q000098:blood supply*; D007511:Ischemia / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000235:genetics / Q000502:physiology*; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D060888:Real-Time Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors
D020413:3' Untranslated Regions; D017931:DNA Primers; D035683:MicroRNAs; C570833:Mirn15 microRNA, mouse; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2