Overexpression of miR-210 and its significance in ischemic tissue damage

Abstract

Hypoxia-induced miR-210 displays a pro-survival, cytoprotective and pro-angiogenic role in several in vitro systems. In vivo, we previously found that miR-210 inhibition increases ischemic damage. Here we describe the generation of a versatile transgenic mouse model allowing the evaluation of miR-210 therapeutic potential in ischemic cardiovascular diseases. We generated a Tet-On miR-210 transgenic mouse strain (TG-210) by targeted transgenesis in the ROSA26 locus. To functionally validate miR-210 transgenic mice, hindlimb ischemia was induced by femoral artery dissection. Blood perfusion was evaluated by power Doppler while tissue damage and inflammation were assessed by histological evaluation. We found that miR-210 levels were rapidly increased in TG-210 mice upon doxycycline administration. miR-210 overexpression was maintained over time and remained within physiological levels in multiple tissues. When hindlimb ischemia was induced, miR-210 overexpression protected from both muscular and vascular ischemic damage, decreased inflammatory cells density and allowed to maintain a better calf perfusion. In conclusion, we generated and functionally validated a miR-210 transgenic mouse model. Albeit validated in the context of a specific cardiovascular ischemic disease, miR-210 transgenic mice may also represent a useful model to assess the function of miR-210 in other physio-pathological conditions.

Hypoxia-Induced miR-210 Is Necessary for Vascular Regeneration upon Acute Limb Ischemia

“…In conclusion, the present study demonstrates that miR-210 plays a pivotal role in post-ischemic neovascularization, suggesting that it could represent a potential therapeutic target in peripheral artery disease. Indeed, miR-210 appears to be a particularly promising one, since its beneficial action is not limited to the early period of ischemic damage when its anti-apoptotic, pro-survival functions can be exploited [6,10,18,19], but also to the following regenerative phases, when its pro-angiogenic action may be harnessed.…”

“…In particular, it can repress mitochondrial metabolism, promoting the shift from mitochondrial respiration to glycolysis [14], inhibiting apoptosis [10,15] and supporting stem-cell survival [16,17]. Accordingly, in limb ischemia, we previously found that miR-210 overexpressing transgenic mice were protected from acute ischemic damage to skeletal muscle and blood vessels [18]. Using a complementary approach, we also found that miR-210 inhibition increased acute skeletal muscle damage with a ROS-mediated mechanism due to insufficient down-regulation of oxidative metabolism upon ischemia [10].…”

“…In the reciprocal gain of function approach, we took advantage of a doxycycline-inducible miR-210 transgenic mouse strain (Tg210) that we had generated and validated in the past [18]. In this case, miR-210 overexpression was induced by doxycycline administration starting from day 4 after surgery and neovascularization was assayed 3 days later.…”

“…Down modulation of miR-210 was carried out by intraperitoneal injection of 12 mg/kg LNA-oligonucleotides against miR-210 (ANTI-210) or a scrambled control sequence (SCR) (In vivo LNAmicroRNA Inhibitors; Exiqon Vedbaek, Denmark) in C57BL/6N male mice (8-12 weeks old, Charles River laboratories, Calco (Lecco), Italy), as previously described [10]. Doxycycline inducible transgenic C57BL/6NTac-Gt(ROSA)26Sor tm3720(Mir210)Tac male mice (Tg210) and Wild Type littermates (WT) were previously described and characterized [18]. When miR-210 expression was induced, WT and Tg210 mice were fed with food containing doxycycline (WT Doxy and Tg210 Doxy ) (Mucedola, Settimo Milanese (MI) Italy, NFM18 diet added with Doxycycline hyclate 2000 mg/kg).…”

Human iPS Cells Derived Skeletal Muscle Progenitor Cells Promote Myoangiogenesis and Restore Dystrophin in Duchenne Muscular Dystrophic Mice

“…Activation of miR-17-92 cluster promoted angiogenesis via PTEN signaling pathway, however, EC miR-17-92 cluster knockout impaired angiogenesis (36). miR-181a and miR-210 are also reported to promote angiogenesis (37)(38)(39)(40). Thus it is very likely that Givi-MPC interacted with resident EC to initiate myogenesis and angiogenesis in Mdx/SCID mice after CTX injury.…”