Comparison of Gene Delivery Techniques for Therapeutic Angiogenesis Ultrasound-Mediated Destruction of Carrier Microbubbles Versus Direct Intramuscular Injection
Comparison of Gene Delivery Techniques for Therapeutic Angiogenesis Ultrasound-Mediated Destruction of Carrier Microbubbles Versus Direct Intramuscular Injection
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DOI：
10.1016/j.jacc.2009.07.023
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发表时间：
2009-10-27
影响因子：
24
通讯作者：
Leong-Poi, Howard
中科院分区：
文献类型：
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作者：
Kobulnik, Jeremy;Kuliszewski, Michael A.;Leong-Poi, Howard
目的 本研究旨在比较通过超声介导 (UM) 破坏静脉内载体微泡与直接肌内 (IM) 注射进行血管生成基因递送的功效。背景 目前血管生成基因治疗的试验仍然受到次优、侵入性递送技术的限制。方法通过结扎 99 只大鼠的髂动脉造成后肢缺血。在 32 只大鼠中，进行了绿色荧光蛋白 (GFP)/血管内皮生长因子 165 (VEGF(165)) 质粒脱氧核糖核酸的 UM 递送。 35 只动物接受了 VEGF(165)/GFP 质粒的肌内注射。其余大鼠不接受任何治疗。分娩前（结扎后第 14 天）以及结扎后第 17、21、28 天和第 8 周，通过对比增强超声评估微血管血容量和流向近端后肢的微血管血流量（每组 n = 8）。通过逆转录酶-聚合酶链式反应评估总转染，并通过免疫组织化学测定转染的定位。结果到第28天，VEGF165的IM和UM递送均产生微血管血容量和微血管血流量的显着增加。虽然治疗组之间微血管血容量的增加相似，但与 IM 治疗的动物相比，UM 治疗的动物的微血管血流量更大 (p < 0.005)，持续到第 8 周。 IM 治疗的动物的 VEGF(165)/GFP 信使核糖核酸表达更大 (p < 0.05)。两组均检测到强 GFP 信号，对于 IM 注射，该信号定位于局灶性血管周围区域和注射部位周围的肌细胞；对于 UM 递送，定位于更广泛分布的小动脉/毛细血管的血管内皮。结论 尽管转染水平较低，但 VEGF(165) 的 UM 递送与 IM 注射一样有效。 UM 递送导致更广泛分布的定向血管转染，这可能是更有效的血管生成的原因。 （J Am Coll Cardiol 2009；54：1735-42）(c) 2009 年，美国心脏病学会基金会
Objectives This study was designed to compare the efficacy of angiogenic gene delivery by ultrasound-mediated (UM) destruction of intravenous carrier microbubbles to direct intramuscular (IM) injections.Background Current trials of gene therapy for angiogenesis remain limited by suboptimal, invasive delivery techniques.Methods Hind-limb ischemia was produced by iliac artery ligation in 99 rats. In 32 rats, UM delivery of green fluorescent protein (GFP)/vascular endothelial growth factor-165 (VEGF(165)) plasmid deoxyribonucleic acid was performed. Thirty-five animals received IM injections of VEGF(165)/GFP plasmid. Remaining rats received no treatment. Before delivery (day 14 after ligation) and at days 17, 21, and 28 and week 8 after ligation, microvascular blood volume and microvascular blood flow to the proximal hind limbs were assessed by contrast-enhanced ultrasound (n = 8 per group). Total transfection was assessed by reverse transcriptase-polymerase chain reaction, and localization of transfection was determined by immunohistochemistry.Results By day 28, both IM and UM delivery of VEGF165 produced significant increases in microvascular blood volume and microvascular blood flow. Whereas increases in microvascular blood volume were similar between treatment groups, microvascular blood flow was greater (p < 0.005) in UM-treated animals as compared with IM-treated animals, persisting to week 8. The VEGF(165)/GFP messenger ribonucleic acid expression was greater (p < 0.05) for IM-treated animals. A strong GFP signal was detected for both groups and was localized to focal perivascular regions and myocytes around injection sites for IM and to the vascular endothelium of arterioles/capillaries in a wider distribution for UM delivery.Conclusions Despite lower transfection levels, UM delivery of VEGF(165) is as effective as IM injections. The UM delivery results in directed vascular transfection over a wider distribution, which may account for the more efficient angiogenesis. (J Am Coll Cardiol 2009; 54: 1735-42) (c) 2009 by the American College of Cardiology Foundation