AAV vectors expressing angiogenic factors for CHD

肌内注射AAV载体是一种有效的递送促红细胞生成素和因子IX等分泌蛋白的途径。因此，这种载体似乎可用于输送治疗冠心病的血管生成因子。初步实验表明，这种管理模式确实是有用的。比较冠状动脉结扎和不结扎小鼠经AAV载体心内注射将VEGF传递到心肌的情况。结扎冠状动脉的小鼠观察到新血管形成，而冠状动脉完整的小鼠观察到很少血管形成。这个项目的第一个目标是将这些研究扩展到更多的动物身上。由于与冠状动脉结扎相关的病理生理变化在大鼠身上得到了更好的证实，因此这些研究将在大鼠身上进行。在处死动物进行病理和血管形成的组织学检查之前，将对动物进行电生理和超声心动图检查。此外，还将使用一种新的心脏血管三维成像方法。(2)本项目的第二个目的是研究调节心肌血管生成因子表达的方法。研究表明，心肌中VEGF的高水平表达可能导致血管瘤的形成，因此控制血管生成因子的表达非常重要。首先可能导致血管瘤的形成，重要的是控制血管生成因子的表达。我们要研究的第一种调节模式我们要研究的是使用促红细胞生成素基因中的缺氧反应元件。初步数据显示，该元件的多个拷贝串联排列可以增加基因的表达，以应对缺氧。其他诱导启动子，如四环素或黄体酮受体系统将被测试。(3)第三个目标是提高新血管形成的质量和数量。血管生成素和成纤维细胞生长因子，单独和联合VEGF将被研究。

Intramuscular injection of AAV vectors is an efficient way of delivering secretory proteins such as erythropoietin and Factor IX. It appears, therefore, that this vector could be useful for delivering angiogenic factors for the treatment of coronary heart disease. Preliminary experiments indicate that this model of administration may indeed by useful. VEGF delivered by AAV vectors into the myocardium by intracardiac injection was compared in mice with or without coronary artery ligation. In mice with ligated coronary arteries, ne blood vessel formation was seen, whereas in the mice with intact coronary, very few blood vessels were seen. (1) The first aim of this project is to extend these studies to a larger number of animals. Because the pathophysiological changes associated with ligation of the coronary artery are much better established in rats, these studies will be performed on rats. The animals will be examined by electrophysiology and echocardiography before they are sacrificed for histological examination for pathology and blood vessel formation. Also, a new method for three dimensional imaging of cardiac blood vessels will be used. (2) A second aim of this project is to investigate methods of regulating angiogenic factors of expression in the myocardium. As it has been shown that high level of expression of VEGF in the myocardium may result in hemangioma formation, it is important to control the expression of angiogenic factors. The first may result in hemangioma formation, it is important to control the expression of angiogenic factors. The first mode of regulation that we will investigate that we will investigate is to use the hypoxia responsive element from the erythropoietin gene. Preliminary data showed several copies of this element arranged in tandem could increase the expression of genes in response to anoxia. Other inducible promoters such as the tetracycline or the progesterone receptor system will be tested. (3) A third aim is to improve the quality and quantity of new blood vessel formation. The angiopoietin and fibroblast growth factor, alone and in combination with VEGF will be investigated.