A selective microRNA-based strategy inhibits restenosis while preserving endothelial function
A selective microRNA-based strategy inhibits restenosis while preserving endothelial function
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DOI：
10.1172/jci76069
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发表时间：
2014-09-01
影响因子：
15.9
通讯作者：
Totary-Jain, Hana
中科院分区：
文献类型：
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作者：
Santulli, Gaetano;Wronska, Anetta;Totary-Jain, Hana
目前批准用于经皮冠状动脉介入治疗的涂层支架的药物不区分增殖的血管平滑肌细胞(VSMCs)和内皮细胞(ECs)。这种歧视的缺乏延迟了血管内皮化和血管愈合，增加了血管成形术后晚期血栓形成的风险。我们开发了一种基于microRNA(基于miRNA)的方法来抑制VSMCs的增殖，从而防止再狭窄，同时选择性地促进再内皮化和保留EC功能。我们使用腺病毒(Ad)载体编码细胞周期蛋白依赖性激酶抑制因子p27(Kip1)(P27)(P27)，在3‘端含有针对EC特异性miR-126-3p的靶序列(Ad-p27-126TS)。用Ad-p27-126T5、Ad-p27(不含miR-126靶序列)或Ad-GFP(对照)分别在体外和大鼠动脉球囊损伤模型中检测外源性p27的过表达。在体外，Ad-p27-126TS保护内皮细胞的增殖、迁移和形成网络的能力。在损伤后2周和4周，Ad-p27-126TS治疗的动物表现出减少的再狭窄，完全的再内皮化，降低的高凝状态，以及对乙酰胆碱的血管扩张反应恢复到与未损伤血管相当的水平。通过掺入miR-126-3p靶序列来利用内源性EC特异性miR-126，我们在VSMC中过度表达外源p27，同时选择性地抑制ECs中p27的过度表达。我们的原则验证研究证明了使用基于miRNA的策略作为一种治疗方法的潜力，以特定地抑制血管再狭窄，同时保留EC功能。
Drugs currently approved to coat stents used in percutaneous coronary interventions do not discriminate between proliferating vascular smooth muscle cells (VSMCs) and endothelial cells (ECs). This lack of discrimination delays reendothelialization and vascular healing, increasing the risk of late thrombosis following angioplasty. We developed a microRNA-based (miRNA-based) approach to inhibit proliferative VSMCs, thus preventing restenosis, while selectively promoting reendothelialization and preserving EC function. We used an adenoviral (Ad) vector that encodes cyclin-dependent kinase inhibitor p27(kip1) (p27) with target sequences for EC-specific miR-126-3p at the 3' end (Ad-p27-126TS). Exogenous p27 overexpression was evaluated in vitro and in a rat arterial balloon injury model following transduction with Ad-p27-126T5, Ad-p27 (without miR-126 target sequences), or Ad-GFP (control). In vitro, Ad-p27-126TS protected the ability of ECs to proliferate, migrate, and form networks. At 2 and 4 weeks after injury, Ad-p27-126TS-treated animals exhibited reduced restenosis, complete reendothelialization, reduced hypercoagulability, and restoration of the vasodilatory response to acetylcholine to levels comparable to those in uninjured vessels. By incorporating miR-126-3p target sequences to leverage endogenous EC-specific miR-126, we overexpressed exogenous p27 in VSMCs, while selectively inhibiting p27 overexpression in ECs. Our proof-of-principle study demonstrates the potential of using a miRNA-based strategy as a therapeutic approach to specifically inhibit vascular restenosis while preserving EC function.