Localized Delivery of shRNA against PHD2 Protects the Heart from Acute Myocardial Infarction through Ultrasound-Targeted Cationic Microbubble Destruction.
Localized Delivery of shRNA against PHD2 Protects the Heart from Acute Myocardial Infarction through Ultrasound-Targeted Cationic Microbubble Destruction.
复制标题
针对 PHD2 的 shRNA 局部递送可通过超声靶向阳离子微泡破坏来保护心脏免受急性心肌梗死的侵害。
作者：
Zhang L;Sun Z;Ren P;You M;Zhang J;Fang L;Wang J;Chen Y;Yan F;Zheng H;Xie M
缺氧诱导因子1α（HIF-1α）在缺血性心脏病中起重要的保护作用。在常氧条件下，HIF-1α被氧依赖性脯氨酰羟化酶-2（PHD 2）降解。基因治疗已成为抑制缺血性损伤后HIF-1α降解、改善心功能的一种有前途的策略。然而，传统的基因递送系统难以实现靶向和局部化的基因递送到缺血心肌中。在这里，我们报告了通过超声靶向微泡破坏（UTMD）局部心肌递送针对PHD 2的shRNA以保护心脏免受急性心肌梗死。本研究利用薄膜水合和超声处理的方法制备了一种新型的阳离子微泡。所得微泡的表面zeta电位为28.2 ± 2.21 mV，每5 × 108个微泡的DNA载量为17.81 ± 1.46 μg，大大提高了DNA的结合性能。结合这些阳离子微泡，UTMD介导的基因递送在H9 C2心肌细胞中进行了评估，并优化了基因转染效率。UTMD介导的shPHD 2转染成功实现了PHD 2基因的敲除，并通过增加HIF-1α、VEGF和bFGF的表达，对H9 C2细胞产生HIF-1α依赖性的保护作用。我们进一步采用UTMD介导的shPHD 2转染到大鼠缺血模型中的局部缺血心肌中，证明梗死面积显着减少，心脏功能显着改善。证实了PHD 2的沉默及其下游基因在处理的心肌中的上调。组织学分析进一步显示shPHD 2处理组中HIF-1α和VEGF以及CD 31阳性细胞数/mm 2显著高于假手术组和对照载体组（P < 0.05）。总之，我们的研究提供了一个很有前途的战略，实现超声介导的心肌局部shRNA递送，以保护心脏免受急性心肌梗死通过阳离子微泡。
Hypoxia-inducible factor 1α (HIF-1α) plays a critical protective role in ischemic heart disease. Under normoxic conditions, HIF-1α was degraded by oxygen-dependent prolyl hydroxylase-2 (PHD2). Gene therapy has become a promising strategy to inhibit the degradation of HIF-1α and to improve cardiac function after ischemic injury. However, conventional gene delivery systems are difficult to achieve a targeted and localized gene delivery into the ischemic myocardia. Here, we report the localized myocardial delivery of shRNA against PHD2 through ultrasound-targeted microbubble destruction (UTMD) for protection the heart from acute myocardial infarction. In this study, a novel cationic microbubble was fabricated by using of the thin-film hydration and sonication method. The resulting microbubbles had a 28.2 ± 2.21 mV surface zeta potential and could greatly improve DNA binding performance, achieving 17.81 ± 1.46 μg of DNA loading capacity per 5 × 108 microbubbles. Combined with these cationic microbubbles, UTMD-mediated gene delivery was evaluated and the gene transfection efficiency was optimized in the H9C2 cardiac cells. Knockdown of PHD2 gene was successfully realized by UTMD-mediated shPHD2 transfection, resulting in HIF-1α-dependent protective effects on H9C2 cells through increasing the expression of HIF-1α, VEGF and bFGF. We further employed UTMD-mediated shPHD2 transfection into the localized ischemic myocardia in a rat ischemia model, demonstrating significantly reduced infarct size and greatly improved the heart function. The silencing of PHD2 and the up-regulation of its downstream genes in the treated myocardia were confirmed. Histological analysis further revealed numbers of HIF-1α- and VEGF-, and CD31-positive cells/mm2 in the shPHD2-treated group were significantly greater than those in the sham or control vector groups (P < 0.05). In conclusion, our study provides a promising strategy to realize ultrasound-mediated localized myocardial shRNA delivery to protect the heart from acute myocardial infarction via cationic microbubbles.
登录
查看更多内容
影响因子：
5.2
作者：
Gandia, Carolina;Arminan, Ana;Sepulveda, Pilar
通讯作者：
Sepulveda, Pilar
影响因子：
10.8
作者：
Otani, Kentaro;Yamahara, Kenichi;Nagaya, Noritoshi
通讯作者：
Nagaya, Noritoshi
影响因子：
9.7
作者：
Nomikou, Nikolitsa;Tiwari, Poonam;McHale, Anthony P.
通讯作者：
McHale, Anthony P.
影响因子：
24
作者：
Kobulnik, Jeremy;Kuliszewski, Michael A.;Leong-Poi, Howard
通讯作者：
Leong-Poi, Howard
影响因子：
9.3
作者：
Lu J;Yao YY;Dai QM;Ma GS;Zhang SF;Cao L;Ren LQ;Liu NF
通讯作者：
Liu NF