ENDOTHELIAL CELL TARGETED GENE THERAPY STRATEGIES FOR CORONARY HEART DISEASE

冠心病的内皮细胞靶向基因治疗策略

• 批准号: 6258937
• 负责人: Victor J Dzau
• 金额: $ 18.3万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6258937
• 关键词: apoptosis; atherosclerosis; enzyme activity; free radical oxygen; gene expression; genetically modified animals; laboratory mouse; laboratory rat; molecular pathology; myocardial ischemia /hypoxia; nitric oxide; nitric oxide synthase; oxidative stress; superoxide dismutase; swine; vascular endothelium

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.

内皮细胞似乎发挥着中心稳态的作用， 维持正常的血管功能和结构。虽然 内皮功能障碍与血管疾病之间存在良好的相关性， 虽然已经建立，但其分子基础和致病意义仍然存在 定义不好。我们的中心假设是正常的内皮细胞 功能取决于反应性和非反应性之间的稳态平衡。 氮物质如一氧化氮（NO）和活性氧物质 (ROS)如超氧阴离子。此外，我们假设，NO- 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包括： 内皮细胞-白细胞粘附，血管平滑肌细胞生长 和迁移，内皮细胞凋亡，血小板聚集， 冠状动脉血流量减少。这个项目将测试假设， 内皮功能障碍特征性NO生物活性下降 是冠状动脉粥样硬化发病机制中的关键事件 心脏病和基因治疗方法的基础上恢复 正常的NO-ROS稳态平衡将是一种新的和有效的长期， 术语治疗方式。我们将集中精力制定一项战略，以“重新- 工程”的内皮细胞，以保持稳态NO-ROS平衡， 使用三种互补的方法：1）增加内皮细胞- 一氧化氮合酶基因表达，以克服增加 2）减少氧化应激介导的NO的释放 通过增加超氧化物歧化酶（一种内源性的 抗氧化剂），以及3）防止功能障碍的出现 表型通过抑制氧化应激诱导的细胞凋亡的周期 和再生功能障碍的细胞通过内皮细胞靶向 抗凋亡基因的表达。这种方法的成功之处在于 基于稳定整合的病毒载体的开发， 静止细胞和血管细胞特异性靶向技术 积极参与这项基因转移计划初始阶段 将利用内皮细胞技术， 在基因工程小鼠中的质量标准，以表征 NO-ROS平衡在动脉粥样硬化形成中的病理生物学意义， 心肌缺血再灌注损伤基于这一特征， 我们将利用该计划中开发的新型病毒载体， 在动物模型的背景下进一步测试这一假设， 模拟人类冠状动脉疾病的治疗。