The use of antibody modified liposomes loaded with AMO-1. to deliver oligonucleotides to ischemic myocardium for arrhythmia therapy
The use of antibody modified liposomes loaded with AMO-1. to deliver oligonucleotides to ischemic myocardium for arrhythmia therapy
复制标题
DOI：
10.1016/j.biomaterials.2013.12.099
复制
发表时间：
2014-04-01
期刊：
影响因子：
14
通讯作者：
Peng, Haisheng
中科院分区：
文献类型：
--
作者：
Liu, Meifang;Li, Minghui;Peng, Haisheng
MicroRNA-1（miR-1）存在于心脏和骨骼组织中。它在缺血性心脏组织中过表达。miR-1反义寡核苷酸（AMO-1）下调miR-1表达可减轻miR-1的促凋亡作用。为了提高AMO-1的治疗效率并抑制其脱靶效应，我们探索了抗心肌肌钙蛋白I（cTnI）抗体修饰的AMO-1脂质体（cT-A-LIP）将寡核苷酸递送至缺血心肌组织。MTT法检测脂质体的细胞毒性。通过体内成像评价对病灶的靶向能力。分别采用活细胞实验台和流式细胞仪观察药物的体外摄取和生物分布。通过心电图、免疫组化、实时荧光PCR和膜片钳记录等方法评价cT-A-LIP的体内抗肿瘤作用。免疫组化显示心肌梗死后第3天cTnI表达达到高峰。cT-LIP经尾静脉给药后，缺血灶中荧光示踪剂的积聚显著增加，高于LIP。此外，cT-A-LIP给药后，缺血性心律失常恢复，ECG ST段抬高接近正常。与MI组相比，MI组miR-1表达明显下调，Kir 2. 1和0：43蛋白表达明显上调。膜片钳记录显示，cT-A-LIP以及AMO 1孵育增加豚鼠心室肌细胞作用于复极化膜电位的K+电流密度。结论：cT-A-LIP不仅能将AMO-1递送至MI大鼠缺血心肌，而且能通过沉默缺血心肌中的miR-1和恢复MI大鼠去极化静息膜电位（RMP）来验证AMO-1对缺血性心律失常的缓解作用。(C)2014爱思唯尔有限公司版权所有。
MicroRNA-1 (miR-1) has been found in cardiac and skeletal tissues. It is overexpressed in ischemic cardiac tissues. Down-regulation of miR-1 could relieve arrhythmogenesis by the anti-miR-1 antisense oligonucleotides (AMO-1). To increase the therapeutic efficiency and inhibit off-target effects of AMO-1, here we explored anti-cardiac troponin I (cTnI) antibody modified liposomes loading with AMO-1 (cT-A-LIP) to deliver the oligonucleotides to ischemic myocardium tissues. Liposomal cytotoxicity was assessed by MTT assay. The targeting abilities to foci were evaluated by in vivo imaging. The uptake and bio-distribution in vitro were observed by live cell station and flow cytomety, respectively. The anti-arrhythmic effects of cT-A-LIP in vivo were evaluated by electrocardiograms (ECG), immunohistochemistry, real-time PCR and patch-clamp recording. Immunohistochemistry showed that cTnI expression had a peak at the third day after myocardial infarction (MI). After cT-LIP administration via tail vein, accumulation of fluorescent trackers in the ischemic foci was significantly increased more than that of LIP. In addition, after cT-A-LIP administration, the ischemic arrhythmias were recovered and ST segment in ECG was elevated nearly back to normal. Compared with MI group, miR-1 expression was significantly down-regulated while Kir2.1 and 0:43 protein expression were increased. Patch-clamp recordings showed that cT-A-LIP as well as AMO1 incubation increased K+ current density in guinea pigs ventricular cardiomyocytes acting on repolarized membrane potential. In conclusion, the cT-A-LIP not only delivered AMO-1 to ischemic myocardium in MI rats, but validated AMO-1 on relieving ischemic arrhythmia by silencing of miR-1 in ischemic myocardium and restoring the depolarized resting membrane potential (RMP) in MI rats. (C) 2014 Elsevier Ltd. All rights reserved.