Efficient myocyte gene delivery with complete cardiac surgical isolation in situ
Efficient myocyte gene delivery with complete cardiac surgical isolation in situ
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DOI：
10.1016/j.jtcvs.2005.07.035
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发表时间：
2005-11-01
影响因子：
6
通讯作者：
Stedman, HH
中科院分区：
文献类型：
--
作者：
Bridges, CR;Gopal, K;Stedman, HH
背景资料：以前，我们使用不完全心脏隔离的体外循环和载体的顺行给药来进行整体心脏基因递送。在这里，我们提出了一个translatable心脏外科手术，允许完全的外科隔离心脏原位逆行（通过冠状静脉循环）管理的载体和内皮透化剂，以增加myocyte transduction efficiency.Methods：在6只成年狗的心脏完全隔离与止血带放置在两个腔静脉和carmulas和所有肺静脉。在心肺转流中，主动脉和肺动脉被交叉夹闭，心脏被分离。将含有10（13）个编码LacZ的腺病毒颗粒和15 μ g血管内皮生长因子的4 ℃晶体心脏停搏液逆行输注到冠状窦中并再循环总共30分钟。然后使犬脱离心肺转流并允许恢复。结果：体外循环组左、右心室β-半乳糖苷酶活性均比对照组高几个数量级（P <0.05）。体外循环组的X-gal染色显示，在相当大比例的心肌细胞中，心肌细胞基因表达的证据明确。结论：本研究为心脏外科手术提供了一种新的技术。这种方法与心脏隔离和逆行交付的载体通过冠状窦的结果在成年大动物体内有效的肌细胞转导。
Background: Previously, we used cardiopulmonary bypass with incomplete cardiac isolation and antegrade administration of vector for global cardiac gene delivery. Here we present a translatable cardiac surgical procedure that allows for complete surgical isolation of the heart in situ with retrograde (through the coronary venous circulation) administration of both vector and endothelial permeabilizing agents to increase myocyte transduction efficiency.Methods: In 6 adult dogs the heart was completely isolated with tourniquets placed around both vena cavae and carmulas and all pulmonary veins. On cardiopulmonary bypass, the aorta and pulmonary artery were crossclamped, and the heart was isolated. Crystalloid cardioplegia at 4 degrees C containing 10(13) particles of adenovirus encoding LacZ and 15 mu g of vascular endothelial growth factor was infused retrograde into the coronary sinus and recirculated for a total of 30 minutes. The dogs were then weaned from cardiopulmonary bypass and allowed to recover. With a catheter, 3 control dogs underwent retrograde infusion of the same cocktail without cardiac isolation or cardiopulmonary bypass.Results: beta-Galactosidase activities in the cardiopulmonary bypass group were several orders of magnitude higher in both the right and left ventricles when compared with those in the control group (P < .05). X-gal staining from the cardiopulmonary bypass group showed unequivocal evidence of myocyte gene expression globally in a significant proportion of cardiac myocytes. No myocyte gene expression was observed in the control group.Conclusion: A novel cardiac surgical technique has been developed. This approach with cardiac isolation and retrograde delivery of vector through the coronary sinus results in efficient myocyte transduction in an adult large animal in vivo.