Therapeutic Angiogenesis by Ultrasound-Mediated MicroRNA-126-3p Delivery
Therapeutic Angiogenesis by Ultrasound-Mediated MicroRNA-126-3p Delivery
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DOI：
10.1161/atvbaha.115.306506
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发表时间：
2015-11-01
影响因子：
8.7
通讯作者：
Leong-Poi, Howard
中科院分区：
文献类型：
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作者：
Cao, Wei J.;Rosenblat, Joshua D.;Leong-Poi, Howard
目的MicroRNA参与包括血管生成在内的多种重要功能。超声靶向微泡破坏（UTMD）是一种用于质粒DNA靶向血管转染的非侵入性技术，并且可能非常适合于促血管生成microRNA递送。我们的目的是调查UTMD的miR-126- 3 p的治疗血管生成在慢性ischemia.Approach和结果miR-126- 3 p的血管生成潜力进行了测试，在人脐静脉内皮细胞在体外。在慢性左股动脉结扎之前和之后，在Fischer-344大鼠中体内测试miR-126- 3 p的UTMD，评估靶敲除、miR-126- 3 p和miR-126- 5 p表达、磷酸化Tie 2水平、微血管灌注和血管密度。在体外，miR-126- 3 p转染的人脐静脉内皮细胞显示出对发芽相关蛋白-1和磷脂酰肌醇-3-激酶调节亚基2的抑制，这些亚基是血管内皮生长因子和血管生成素-1信号的负调节剂，通过敲低磷脂酰肌醇-3-激酶调节亚基2介导的磷酸化Tie 2增加，以及通过血管内皮生长因子/血管生成因子介导的血管生成潜能增加。血管内皮生长因子R2和血管生成素-1/Tie2的作用。miR-126- 3 p的UTMD导致靶向血管转染，在递送后早期达到峰值并持续>3天，并导致发芽相关蛋白-1和磷脂酰肌醇-3-激酶调节亚基2的抑制，在远端器官中的摄取最小。最后，miR-126- 3 p对慢性缺血后肢肌肉的UTMD导致改善的灌注、血管密度、增强的小动脉形成、周细胞覆盖和磷酸化Tie 2水平，而不影响miR-126- 5 p或delta-like 1同源物水平。结论miR-126的UTMD通过抑制发芽相关蛋白，改善慢性缺血背景下的组织灌注和血管密度。1和磷脂酰肌醇-3-激酶调节亚基2，增强血管内皮生长因子和血管生成素-1信号传导，对miR-126- 5 p没有影响。UTMD是一种有前途的microRNA递送平台，可用于治疗性血管生成。
Objective MicroRNAs are involved in many critical functions, including angiogenesis. Ultrasound-targeted microbubble destruction (UTMD) is a noninvasive technique for targeted vascular transfection of plasmid DNA and may be well suited for proangiogenic microRNA delivery. We aimed to investigate UTMD of miR-126-3p for therapeutic angiogenesis in chronic ischemia.Approach and Results The angiogenic potential of miR-126-3p was tested in human umbilical vein endothelial cells in vitro. UTMD of miR-126-3p was tested in vivo in Fischer-344 rats before and after chronic left femoral artery ligation, evaluating target knockdown, miR-126-3p and miR-126-5p expression, phosphorylated Tie2 levels, microvascular perfusion, and vessel density. In vitro, miR-126-3p-transfected human umbilical vein endothelial cells showed repression of sprouty-related protein-1 and phosphatidylinositol-3-kinase regulatory subunit 2, negative regulators of vascular endothelial growth factor and angiopoietin-1 signaling, increased phosphorylated Tie2 mediated by knockdown of phosphatidylinositol-3-kinase regulatory subunit 2 and greater angiogenic potential mediated by both vascular endothelial growth factor/vascular endothelial growth factor R2 and angiopoietin-1/Tie2 effects. UTMD of miR-126-3p resulted in targeted vascular transfection, peaking early after delivery and lasting for >3 days, and resulting in inhibition of sprouty-related protein-1 and phosphatidylinositol-3-kinase regulatory subunit 2, with minimal uptake in remote organs. Finally, UTMD of miR-126-3p to chronic ischemic hindlimb muscle resulted in improved perfusion, vessel density, enhanced arteriolar formation, pericyte coverage, and phosphorylated Tie2 levels, without affecting miR-126-5p or delta-like 1 homolog levels.Conclusions UTMD of miR-126 results in improved tissue perfusion and vascular density in the setting of chronic ischemia by repressing sprouty-related protein-1 and phosphatidylinositol-3-kinase regulatory subunit 2 and enhancing vascular endothelial growth factor and angiopoietin-1 signaling, with no effect on miR-126-5p. UTMD is a promising platform for microRNA delivery, with applications for therapeutic angiogenesis.