Hypoxia-responsive microRNA-101 promotes angiogenesis via heme oxygenase-1/vascular endothelial growth factor axis by targeting cullin 3.
OBJECTIVE
Hypoxia induces expression of various genes and microRNAs (miRs) that regulate angiogenesis and vascular function. In this study, we investigated a new functional role of new hypoxia-responsive miR-101 in angiogenesis and its underlying mechanism for regulating heme oxygenase-1 (HO-1) and vascular endothelial growth factor (VEGF) expression.


RESULTS
We found that hypoxia induced miR-101, which binds to the 3'untranslated region of cullin 3 (Cul3) and stabilizes nuclear factor erythroid-derived 2-related factor 2 (Nrf2) via inhibition of the proteasomal degradation pathway. miR-101 overexpression promoted Nrf2 nuclear accumulation, which was accompanied with increases in HO-1 induction, VEGF expression, and endothelial nitric oxide synthase (eNOS)-derived nitric oxide (NO) production. The elevated NO-induced S-nitrosylation of Kelch-like ECH-associated protein 1 and subsequent induction of Nrf2-dependent HO-1 lead to further elevation of VEGF production via a positive feedback loop between the Nrf2/HO-1 and VEGF/eNOS axes. Moreover, miR-101 promoted angiogenic signals and angiogenesis both in vitro and in vivo, and these events were attenuated by inhibiting the biological activity of HO-1, VEGF, or eNOS. Moreover, these effects were also observed in aortic rings from HO-1(+/-) and eNOS(-/-) mice. Local overexpression of miR-101 improved therapeutic angiogenesis and perfusion recovery in the ischemic mouse hindlimb, whereas antagomiR-101 diminished regional blood flow.


METHODS
Hypoxia-responsive miR-101 stimulates angiogenesis by activating the HO-1/VEGF/eNOS axis via Cul3 targeting. Thus, miR-101 is a novel angiomir.


CONCLUSIONS
Our results provide new mechanistic insights into a functional role of miR-101 as a potential therapeutic target in angiogenesis and vascular remodeling.
1 Department of Molecular and Cellular Biochemistry, Kangwon National University , Chuncheon, South Korea .
Ji-Hee
JH
Kim
Kwang-Soon
KS
Lee
Dong-Keon
DK
Lee
Joohwan
J
Kim
Su-Nam
SN
Kwak
Kwon-Soo
KS
Ha
Jongseon
J
Choe
Moo-Ho
MH
Won
Byung-Ryul
BR
Cho
Dooil
D
Jeoung
Hansoo
H
Lee
Young-Guen
YG
Kwon
Young-Myeong
YM
Kim
D000818:Animals; D015687:Cell Hypoxia / Q000235:genetics; D044842:Cullin Proteins / Q000235:genetics* / Q000378:metabolism; D016228:Endothelial Growth Factors / Q000235:genetics / Q000378:metabolism; D051547:Heme Oxygenase-1 / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D051267:NF-E2-Related Factor 2 / Q000235:genetics; D018919:Neovascularization, Physiologic / Q000235:genetics*; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
C503646:Cul3 protein, mouse; D044842:Cullin Proteins; D016228:Endothelial Growth Factors; C581233:MIRN101 microRNA, mouse; D035683:MicroRNAs; D051267:NF-E2-Related Factor 2; C495636:Nfe2l2 protein, mouse; D042461:Vascular Endothelial Growth Factor A; C467480:vascular endothelial growth factor A, mouse; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse; D051547:Heme Oxygenase-1