From the Cardiovascular Division, Department of Medicine (B.L., W.W., D.O., H.L., H.S.C., S.H., X.L., S.N.A., N.L., I.H., K.C., M.W.F.), Brigham and Women's Hospital, Harvard Medical School, Boston, MA.
Department of Medical Biology, Hacettepe University, Ankara, Turkey (D.O.).
Arterioscler Thromb Vasc Biol. 2019 Jul;39(7):1458-1474. doi: 10.1161/ATVBAHA.119.312726. Epub 2019 May 16.
Objective- In response to tissue injury, the appropriate progression of events in angiogenesis is controlled by a careful balance between pro and antiangiogenic factors. We aimed to identify and characterize microRNAs that regulate angiogenesis in response to tissue injury. Approach and Results- We show that in response to tissue injury, microRNA-615-5p (miR-615-5p) is rapidly induced and serves as an antiangiogenic microRNA by targeting endothelial cell VEGF (vascular endothelial growth factor)-AKT (protein kinase B)/eNOS (endothelial nitric oxide synthase) signaling in vitro and in vivo. MiR-615-5p expression is increased in wounds of diabetic db/db mice, in plasma of human subjects with acute coronary syndromes, and in plasma and skin of human subjects with diabetes mellitus. Ectopic expression of miR-615-5p markedly inhibited endothelial cell proliferation, migration, network tube formation in Matrigel, and the release of nitric oxide, whereas miR-615-5p neutralization had the opposite effects. Mechanistic studies using transcriptomic profiling, bioinformatics, 3' untranslated region reporter and microribonucleoprotein immunoprecipitation assays, and small interfering RNA dependency studies demonstrate that miR-615-5p inhibits the VEGF-AKT/eNOS signaling pathway in endothelial cells by targeting IGF2 (insulin-like growth factor 2) and RASSF2 (Ras-associating domain family member 2). Local delivery of miR-615-5p inhibitors, markedly increased angiogenesis, granulation tissue thickness, and wound closure rates in db/db mice, whereas miR-615-5p mimics impaired these effects. Systemic miR-615-5p neutralization improved skeletal muscle perfusion and angiogenesis after hindlimb ischemia in db/db mice. Finally, modulation of miR-615-5p expression dynamically regulated VEGF-induced AKT signaling and angiogenesis in human skin organoids as a model of tissue injury. Conclusions- These findings establish miR-615-5p as an inhibitor of VEGF-AKT/eNOS-mediated endothelial cell angiogenic responses and that manipulating miR-615-5p expression could provide a new target for angiogenic therapy in response to tissue injury. Visual Overview- An online visual overview is available for this article.
目的-在组织损伤的情况下，血管生成过程中事件的适当进展受到促血管生成和抗血管生成因子之间的精细平衡的控制。我们旨在鉴定和表征调节组织损伤后血管生成的 microRNA。
方法和结果-我们表明，在组织损伤的情况下，microRNA-615-5p（miR-615-5p）被迅速诱导，并通过靶向体外和体内内皮细胞 VEGF（血管内皮生长因子）-AKT（蛋白激酶 B）/eNOS（内皮型一氧化氮合酶）信号传导作为抗血管生成 microRNA。在糖尿病 db/db 小鼠的伤口、急性冠状动脉综合征患者的血浆以及糖尿病患者的血浆和皮肤中，miR-615-5p 的表达增加。外源性表达 miR-615-5p 显著抑制内皮细胞增殖、迁移、Matrigel 网络管形成和一氧化氮释放，而 miR-615-5p 中和则产生相反的效果。使用转录组谱分析、生物信息学、3'非翻译区报告和 microribonucleoprotein 免疫沉淀测定以及小干扰 RNA 依赖性研究的机制研究表明，miR-615-5p 通过靶向 IGF2（胰岛素样生长因子 2）和 RASSF2（Ras 相关结构域家族成员 2）抑制内皮细胞中的 VEGF-AKT/eNOS 信号通路。在 db/db 小鼠中局部递送 miR-615-5p 抑制剂可显著增加血管生成、肉芽组织厚度和伤口闭合率，而 miR-615-5p 模拟物则会降低这些作用。全身性 miR-615-5p 中和可改善 db/db 小鼠后肢缺血后的骨骼肌灌注和血管生成。最后，在组织损伤模型人皮肤类器官中，miR-615-5p 表达的调节动态调节了 VEGF 诱导的 AKT 信号和血管生成。
结论-这些发现确立了 miR-615-5p 作为 VEGF-AKT/eNOS 介导的内皮细胞血管生成反应的抑制剂，并且操纵 miR-615-5p 表达可能为组织损伤后血管生成治疗提供新的靶标。
可视化概述-本文提供了一个在线可视化概述。
