ENDOTHELIAL CELL TARGETED GENE THERAPY STRATEGIES FOR CORONARY HEART DISEASE

血管内皮细胞似乎扮演着中枢内环境平衡的角色。 维持正常的血管功能和结构。尽管 血管内皮细胞功能障碍与血管疾病之间的相关性很好 它的分子基础和致病意义仍然存在 定义不明确。我们的中心假设是正常的内皮细胞 功能是基于体内平衡和反应性之间的平衡 氮物种，如一氧化氮(NO)和活性氧物种 (ROS)，如超氧阴离子。此外，我们假设没有- ROS平衡调节几个关键的病理生物学过程 参与动脉粥样硬化性冠心病的因素包括： 内皮细胞-白细胞黏附，血管平滑肌细胞生长 迁移，内皮细胞凋亡，血小板聚集，以及 冠脉血流量减少。这个项目将检验这一假设 血管内皮细胞功能障碍的NO生物活性下降 是动脉粥样硬化性冠状动脉病变发病机制中的关键事件 心脏病和基于恢复心脏的基因治疗方法 正常的NO-ROS动态平衡将是一种新颖而有效的长效平衡 定期治疗方式。我们将专注于制定一项战略，以“重新-- 工程“内皮细胞，以维持动态平衡的NO-ROS平衡 使用三种互补的方法：1)增强内皮细胞- 一氧化氮合酶基因的表达将覆盖增加的 NO的分解代谢，2)减少氧化应激介导的NO分解代谢 通过增强超氧化物歧化酶(一种内源性 抗氧化剂)，以及3)防止功能障碍的出现 抑制氧化应激诱导的细胞凋亡循环的表型 通过内皮细胞靶向再生功能障碍的细胞 抗细胞凋亡基因的表达。这种方法的成功之处在于 基于稳定整合的病毒载体的发展 静止细胞和血管细胞特异性靶向技术 积极参与该基因转移项目的研究。初级阶段 该项目将利用内皮细胞技术 在基因工程小鼠中的规范来表征 NO-ROS平衡在动脉粥样硬化发生和发展中的病理生物学意义 心肌缺血再灌注损伤。基于这一特征， 我们将利用在该计划内开发的新型病毒载体来 在动物模型的背景下进一步检验这一假设 模拟人类冠状动脉疾病的治疗。

The endothelium appears to play a central homeostatic role in maintaining normal vascular function and structure. Although the association between endothelial dysfunction and vascular disease is well established, its molecular basis and pathogenic significance remains poorly defined. Our central hypothesis poses that normal endothelial function is predicated upon a homeostatic balance between reactive nitrogen species such as nitric oxide (NO) and reactive oxygen species (ROS) such as superoxide anion. Furthermore, we postulate that the NO- ROS balance modulates several critical pathobiological processes involved in atherosclerotic coronary heart disease including : endothelial cell-leukocyte adhesion, vascular smooth muscle cell growth and migration, endothelial cell apoptosis, platelet aggregation, and decreased coronary blood flow. This project will test the postulate that the decline in NO bioactivity characteristic of endothelial dysfunction is a critical event in the pathogenesis of atherosclerotic coronary heart disease and that a gene therapy approach based upon restoring the normal NO-ROS homeostatic balance will be a novel and effective long- term treatment modality. We will focus on developing a strategy to "re- engineer" the endothelium to preserve the homeostatic NO-ROS balance by using three complementary approaches : 1) augment endothelial cell- nitric oxide synthase gene expression to override the increased catabolism of NO, 2) decrease oxidative stress-mediated catabolism of NO by augmenting the expression of superoxide dismutase (an endogenous anti-oxidant), and 3) prevent the emergence of the dysfunctional phenotype by inhibiting the cycle of oxidative stress-induced apoptosis and regeneration of dysfunctional cells by endothelial cell-targeted expression of anti-apoptotic genes. The success of this approach is predicated upon the development of stably integrating viral vectors for quiescent cells and vascular cell-specific targeting technologies under active investigation in this Gene Transfer Program. The initial phases of the project will utilize technologies of endothelial cell specification in genetically engineered mice to characterize the pathobiological significance of the NO-ROS balance in atherogenesis and cardiac ischemia-reperfusion injury. Based upon this characterization, we will utilize the novel viral vectors developed within the Program to further test this hypothesis in the context of animal models that simulate the treatment of human coronary artery disease.