MicroRNA Detargeting Novel Therapy for Coronary Artery Disease

7.项目摘要/摘要 心血管疾病占全球死亡人数的近三分之一，而冠状动脉疾病 仍然是美国头号死因。2002年药物洗脱支架(DES)介绍 与裸机相比，通过显著降低再狭窄率，使PCI领域发生革命性变化 支架(BMS)。尽管DES具有明显的临床优势，但人们对其长期疗效表示担忧。 安全性，特别是与延迟内皮化相关的支架血栓形成。新疗法 需要针对VSMC和其他浸润性细胞而不是VEC的策略。 MiRNAs通过抑制翻译或促进基因表达来作为基因表达的负调控因子 靶基因的降解。最近的研究表明，mir-143/145星系团在 SMC在静止期和增殖期之间的表型转换。这些发现打开了 为治疗再狭窄打开了潜在的新疗法之门。然而，我们的初步数据显示，增加 这些miRNAs的非靶向表达可抑制血管内皮细胞的生长和迁移。因此，在这个 我们建议的应用：1)研究MIR-143/145去靶向策略对VSMC和VEC的影响 体外增殖和迁移。2)评估去靶化策略的特异度和敏感性 大鼠颈动脉球囊损伤的体内实验，以及3)在相同的 活体球囊损伤模型。为了实现这些目标，我们将使用内皮细胞特异的miRNA来 优势(mir-126)，我们将mir-126的目标序列或随机序列插入到 表达慢病毒的MIR-143/mir-145。被感染的VSMC和VEC将接受增殖和迁移测试。 同样的病毒将在球囊损伤后立即注射给大鼠，并 将评估新生内膜/中膜比率和血管内皮细胞的完整性。 此研究计划是职业发展计划的一部分，申请者通过该计划旨在 获得关键知识和技术技能。非凡的设施和可用的顾问和 合作者确保培训期间的最佳环境。因此，指导阶段将允许 成功地过渡到独立阶段。申请者将继续制定以下最终目标 这项研究项目的长期目标是开发治疗冠状动脉疾病的新的靶向疗法 动脉疾病。

7. Project Summary/Abstract Cardiovascular disease accounts for nearly one third of deaths globally, and Coronary Artery Disease remains the #1 cause of death in the United States. The introduction of the Drug-Eluting Stent (DES) in 2002 revolutionized the field of PCI, by significantly reducing rates of restenosis when compared to bare-metal stents (BMS). Despite the clear clinical advantage of DES, concerns have been raised over their long-term safety, with particular reference to stent thrombosis related to delayed endothelialisation. New therapeutic strategies that can specifically target VSMC and other infiltrated cells but not VEC are needed. MiRNAs act as negative regulators of gene expression by inhibiting the translation or promoting the degradation of target mRNAs. Recent studies show that the mir-143/145 cluster plays important roles in the phenotypic switching of SMC between the quiescent and the proliferative phenotypes. These findings opened the door to potentially novel therapies for restenosis. However, our preliminary data show that increasing the expression of these miRNAs in a non-targeted manner inhibits VEC growth and migration. Therefore, in this application we propose to: 1) Investigate the effect of a mir-143/145 de-targeting strategy on VSMC and VEC proliferation and migration in vitro. 2) Evaluate the specificity and the sensitivity of the de-targeting strategy in a rat carotid artery balloon injury in vivo, and 3) Examine the mir-143/145 de-targeting strategy in the same balloon injury model in vivo. To achieve these aims we will use the endothelial cell specific miRNA to our advantage (mir-126), and we will insert target sequences for mir-126 or random sequences into the 3'-end of a mir-143/mir-145 expressing lentivirus. Infected VSMC and VEC will be tested for proliferation and migration. The same viruses will be will be administered to the rats immediately following balloon injury and the neointimal/media ratio and the integrity of VEC will be assessed. This research plan is meant to be part of a Career Development Plan through which the applicant aims to obtain critical knowledge and technical skills. The extraordinary facilities and the availability of advisors and collaborators ensure an optimal environment for the training period. The mentored phase will therefore allow the transition with success to an independent phase. The applicant will continue to develop the final aims of this research project with the long-term goal to develop novel targeted therapies for the treatment of Coronary Artery Disease.