MicroRNA Detargeting Novel Therapy for Coronary Artery Disease

描述(申请人提供)：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的头号死因。2002年推出的药物洗脱支架(DES)与裸金属支架(BMS)相比，显著降低了再狭窄率，彻底改变了经皮冠状动脉介入治疗(PCI)领域。尽管DES具有明显的临床优势，但人们对其长期安全性提出了担忧，尤其是与延迟内皮化相关的支架血栓形成。需要针对VSMC和其他浸润性细胞而不是VEC的新的治疗策略。MiRNAs通过抑制靶mRNAs的翻译或促进其降解，作为基因表达的负调控因子。最近的研究表明，mir-143/145簇在SMC从静止表型到增殖表型的表型转换中起着重要作用。这些发现为潜在的治疗再狭窄的新方法打开了大门。然而，我们的初步数据显示，以非靶向的方式增加这些miRNAs的表达会抑制血管内皮细胞的生长和迁移。因此，在这一应用中，我们建议：1)研究去靶向策略mir-143/145对VSMC和VEC体外增殖和迁移的影响。2)评价去靶向策略在大鼠颈动脉球囊损伤中的特异性和敏感性，以及3)在相同的球囊损伤模型中检测mir-143/145去靶向策略。为了达到这些目的，我们将利用内皮细胞特异性的miRNA(mir-126)，并将mir-126的靶序列或随机序列插入到表达mir-143/mir-145的慢病毒的3‘端。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/中层比率和血管内皮细胞的完整性。这项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。非同寻常的设施以及顾问和合作者的提供确保了培训期间的最佳环境。因此，指导阶段将允许成功过渡到独立阶段。申请者将继续开发这一研究项目的最终目标，长期目标是开发治疗冠状动脉疾病的新的靶向疗法。

DESCRIPTION (provided by applicant): 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 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.