Departamento de Farmacología, Facultad de Medicina, Universidad de Valencia, Spain.
J Gene Med. 2010 Nov;12(11):920-6. doi: 10.1002/jgm.1510. Epub 2010 Oct 22.
Hydrodynamic injection has demonstrated to be very efficient in the liver of small animals, although this procedure must be translated to the clinical practice in a milder but no less efficient way. The present study evaluates the capacity of non-invasive interventional catheterization as a procedure for naked DNA delivery to the heart in large animals.
Two catheters were placed in the coronary sinus: one of them to block blood circulation and the other to retrogradely inject 50 ml of a saline solution of DNA (20 µg/ml) containing the enhanced green fluorescent protein (EGFP) gene, at a flow rate of 5 ml/s.
The results obtained show that EGFP protein, identified by immunohistochemistry, was present and widely distributed throughout the atrial and ventricular cardiac tissue. This observation agrees with the efficiency of EGFP gene delivery resulting in 1-200 EGFP gene copies per endogenous haploid genome. However, the transcription efficiency of the exogenous EGFP gene was at a ratio of 0.2-10 copies with respect to the endogenous GAPDH gene, suggesting that optimized gene constructs for expression in cardiac tissue could increase the final efficacy of gene transfer.
We conclude that the retrovenous injection of naked DNA in the coronary sinus employing the catheterization technique is an easy and probably safe method for whole cardiac gene transfer.
在小动物的肝脏中，流体动力学注射已被证明非常有效，尽管该程序必须以更温和但同样有效的方式转化为临床实践。本研究评估了非侵入性介入导管术作为将裸 DNA 递送至大型动物心脏的方法的能力。
在冠状窦中放置了两个导管：一个用于阻断血液循环，另一个用于以 5ml/s 的流速逆行注射 50ml 含有增强型绿色荧光蛋白 (EGFP) 基因的 DNA（20µg/ml）生理盐水溶液。
获得的结果表明，通过免疫组织化学鉴定，存在并广泛分布于心房和心室心肌组织中的 EGFP 蛋白。这一观察结果与 EGFP 基因传递的效率一致，导致每个内源性单倍体基因组中有 1-200 个 EGFP 基因拷贝。然而，外源性 EGFP 基因的转录效率与内源性 GAPDH 基因的比例为 0.2-10 拷贝，这表明用于心脏组织表达的优化基因构建体可以提高基因转移的最终效率。
我们得出结论，采用导管技术经冠状窦逆行注射裸 DNA 是一种简单且可能安全的心脏整体基因转移方法。
