Hypoxia-induced miR-210 modulates tissue response to acute peripheral ischemia.
OBJECTIVE
Peripheral artery disease is caused by the restriction or occlusion of arteries supplying the leg. Better understanding of the molecular mechanisms underpinning tissue response to ischemia is urgently needed to improve therapeutic options. The aim of this study is to investigate hypoxia-induced miR-210 regulation and its role in a mouse model of hindlimb ischemia.


RESULTS
miR-210 expression was induced by femoral artery dissection. To study the role of miR-210, its function was inhibited by the systemic administration of a miR-210 complementary locked nucleic acid (LNA)-oligonucleotide (anti-miR-210). In the ischemic skeletal muscle, anti-miR-210 caused a marked decrease of miR-210 compared with LNA-scramble control, while miR-210 target expression increased accordingly. Histological evaluation of acute tissue damage showed that miR-210 inhibition increased both apoptosis at 1 day and necrosis at 3 days. Capillary density decrease caused by ischemia was significantly more pronounced in anti-miR-210-treated mice; residual limb perfusion decreased accordingly. To investigate the molecular mechanisms underpinning the increased damage triggered by miR-210 blockade, we tested the impact of anti-miR-210 treatment on the transcriptome. Gene expression analysis highlighted the deregulation of mitochondrial function and redox balance. Accordingly, oxidative damage was more severe in the ischemic limb of anti-miR-210-treated mice and miR-210 inhibition increased oxidative metabolism. Further, oxidative-stress resistant p66(Shc)-null mice displayed decreased tissue damage following ischemia.


METHODS
This study identifies miR-210 as a crucial element in the adaptive mechanisms to acute peripheral ischemia.


CONCLUSIONS
The physiopathological significance of miR-210 is context dependent. In the ischemic skeletal muscle it seems to be cytoprotective, regulating oxidative metabolism and oxidative stress.
1 Molecular Cardiology Laboratory, IRCCS-Policlinico San Donato , Milan, Italy .
Germana
G
Zaccagnini
Biagina
B
Maimone
Valeria
V
Di Stefano
Pasquale
P
Fasanaro
Simona
S
Greco
Alessandra
A
Perfetti
Maurizio C
MC
Capogrossi
Carlo
C
Gaetano
Fabio
F
Martelli
D000208:Acute Disease; D000818:Animals; D017209:Apoptosis; D015687:Cell Hypoxia; D002460:Cell Line; D015870:Gene Expression; D006019:Glycolysis; D006614:Hindlimb / Q000098:blood supply; D007511:Ischemia / Q000235:genetics / Q000378:metabolism*; D008297:Male; D057507:Mice, 129 Strain; D018345:Mice, Knockout; D035683:MicroRNAs / Q000502:physiology*; D018482:Muscle, Skeletal / Q000098:blood supply / Q000378:metabolism / Q000473:pathology; D018384:Oxidative Stress; D034622:RNA Interference
C549325:MIRN210 microRNA, mouse; D035683:MicroRNAs