Direct in vivo gene transfer into the coronary and peripheral vasculatures of the intact dog.
Direct in vivo gene transfer into the coronary and peripheral vasculatures of the intact dog.
复制标题
将体内基因直接转移到完整狗的冠状动脉和外周脉管系统中。
DOI：
10.1161/01.cir.83.6.2007
复制
发表时间：
1991
期刊：
影响因子：
37.8
通讯作者：
Swain,JL
中科院分区：
文献类型：
--
作者：
Lim,CS;Chapman,GD;Gammon,RS;Muhlestein,JB;Bauman,RP;Stack,RS;Swain,JL
基因治疗方法已被建议用于治疗心血管疾病。最近，已证实将编码β-半乳糖苷酶的基因直接转移到猪的外周动脉中。为了确定这种方法是否适用于其他动脉床和其他物种，我们首先在犬模型中评估了β-半乳糖苷酶作为标记蛋白的使用。我们证明，可变的，但大量的内源性β-半乳糖苷酶样活性诱导的犬外周动脉，这排除了使用这种标记蛋白在该模型中的基因转移的效率进行评估的操作。然后评价编码萤火虫荧光素酶的标记基因，发现即使在动脉操作后，背景荧光素酶活性在狗中也很低。使用荧光素酶基因，我们然后证明脂质介导的基因转移直接进入冠状动脉和外周动脉的完整的狗。这些结果表明了体内基因转移到冠状动脉中的可行性，并证明了在犬模型中基因转移后荧光素酶标记蛋白在定量重组蛋白表达中的用途。这种简单有效的体内基因转移到冠状动脉和外周动脉的方法可能适用于局部生产治疗重要的蛋白质治疗心血管疾病。
Gene therapy approaches have been suggested for the treatment of cardiovascular disease. Recently, direct transfer of the gene encoding beta-galactosidase into peripheral arteries of the pig has been demonstrated. To determine whether this approach is applicable to other arterial beds and to other species, we first evaluated the use of beta-galactosidase as a marker protein in the canine model. We demonstrate that variable but substantial endogenous beta-galactosidase-like activity is induced by manipulation of canine peripheral arteries, which precludes the use of this marker protein in evaluating the efficiency of gene transfer in this model. A marker gene encoding firefly luciferase was then evaluated, and background luciferase activity was found to be low in the dog even after arterial manipulation. Using the luciferase gene, we then demonstrated lipid-mediated gene transfer directly into both coronary and peripheral arteries of the intact dog. These results indicate the feasibility of in vivo gene transfer into coronary arteries and demonstrate the use of the luciferase marker protein in quantifying recombinant protein expression following gene transfer in canine models. This simple and effective method for direct in vivo gene transfer into coronary and peripheral arteries may be applicable to the localized production of therapeutically important proteins for the treatment of cardiovascular diseases.