Asymmetric Dimethylarginine Impairs Angiogenic Progenitor Cell Function in Patients With Coronary Artery Disease Through a MicroRNA-21-Dependent Mechanism
Asymmetric Dimethylarginine Impairs Angiogenic Progenitor Cell Function in Patients With Coronary Artery Disease Through a MicroRNA-21-Dependent Mechanism
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DOI：
10.1161/circresaha.110.216770
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发表时间：
2010-07-09
影响因子：
20.1
通讯作者：
Thum, Thomas
中科院分区：
文献类型：
--
作者：
Fleissner, Felix;Jazbutyte, Virginija;Thum, Thomas
基本原理：内源性一氧化氮合酶抑制剂不对称二甲基精氨酸（ADMA）在冠心病患者中增加，并且可能通过小的调节RNA调节循环血管生成祖细胞（APCs）的功能。目的：研究microRNA在ADMA介导的APCs损伤中的作用。方法和结果：通过使用微阵列分析，我们建立了人APCs的microRNA表达谱。我们使用ADMA诱导APC功能障碍，发现16个失调的microRNA。我们专注于miR-21，其通过ADMA处理上调3倍。miR-21在人APC中的过表达损害了迁移能力。为了鉴定受调控的miR-21靶标，我们使用蛋白质组分析，使用差异凝胶电泳，然后对受调控的蛋白质进行质谱分析。我们发现miR-21前体的转染显著抑制APC中的超氧化物歧化酶2，这导致细胞内活性氧浓度增加和一氧化氮生物利用度受损。miR-21进一步抑制sprouty-2，导致Erk Map激酶依赖性活性氧簇形成和APC迁移缺陷。小干扰RNA介导的超氧化物歧化酶2或发芽-2减少也增加了活性氧的形成和APC迁移能力受损。ADMA介导的活性氧簇形成和APC功能障碍通过miR-21阻断而得以挽救。来自冠状动脉疾病和高ADMA血浆水平患者的APC显示>4倍的miR-21水平升高，低超氧化物歧化酶2表达，和受损的迁移能力，这可以通过miR-21拮抗剂正常化。结论：我们确定了一种新的miR-21依赖机制ADMA介导的APC功能障碍。因此，miR-21拮抗作用成为改善冠状动脉疾病患者中功能障碍的APC的有趣策略。(Circ Res. 2010; 107：138-143）。
Rationale: The endogenous nitric oxide synthase inhibitor asymmetrical dimethylarginine (ADMA) is increased in patients with coronary artery disease and may regulate function of circulating angiogenic progenitor cells (APCs) by small regulatory RNAs.Objectives: To study the role of microRNAs in ADMA-mediated impairment of APCs.Methods and Results: By using microarray analyses, we established microRNA expression profiles of human APCs. We used ADMA to induce APC dysfunction and found 16 deregulated microRNAs. We focused on miR-21, which was 3-fold upregulated by ADMA treatment. Overexpression of miR-21 in human APCs impaired migratory capacity. To identify regulated miR-21 targets, we used proteome analysis, using difference in-gel electrophoresis followed by mass spectrometric analysis of regulated proteins. We found that transfection of miR-21 precursors significantly repressed superoxide dismutase 2 in APCs, which resulted in increased intracellular reactive oxygen species concentration and impaired nitric oxide bioavailability. MiR-21 further repressed sprouty-2, leading to Erk Map kinase-dependent reactive oxygen species formation and APC migratory defects. Small interference RNA-mediated superoxide dismutase 2 or sprouty-2 reduction also increased reactive oxygen species formation and impaired APC migratory capacity. ADMA-mediated reactive oxygen species formation and APC dysfunction was rescued by miR-21 blockade. APCs from patients with coronary artery disease and high ADMA plasma levels displayed >4-fold elevated miR-21 levels, low superoxide dismutase 2 expression, and impaired migratory capacity, which could be normalized by miR-21 antagonism.Conclusions: We identified a novel miR-21-dependent mechanism of ADMA-mediated APC dysfunction. MiR-21 antagonism therefore emerges as an interesting strategy to improve dysfunctional APCs in patients with coronary artery disease. (Circ Res. 2010; 107: 138-143.)