Blocking Intimal Hyperplasia Following Vascular Trauma

阻止血管创伤后的内膜增生

• 批准号: 6537296
• 负责人: Edith Tzeng
• 金额: $ 29.01万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6537296
• 关键词: Adenoviridae; G protein; Retroviridae; atherosclerosis; cardiovascular disorder prevention; cardiovascular injury; cell proliferation; disease /disorder model; enzyme induction /repression; gene expression; gene therapy; genetic transduction; hypercholesterolemia; hyperplasia; laboratory rat; mitogen activated protein kinase; nitric oxide; nitric oxide synthase; prostaglandin endoperoxide synthase; protein structure function; restenosis; swine; transfection /expression vector; trauma; vascular endothelium; vascular smooth muscle

Atherosclerotic vascular disease is the leading cause of morbidity and mortality in the United States. Conventional treatments such as angioplasty and bypass procedures are limited in success by the development of intimal hyperplasia (IH). Over the last decade, gene therapy approaches to treating IH have been developed. We propose that one of the best candidate genes for the prevention of IH is the inducible nitric oxide synthase (iNOS) gene. iNOS produces NO which can inhibit smooth muscle cell (SMC) proliferation, promote endothelial cell (EC) regrowth, inhibit platelet and leukocyte adhesion, and promote vasodilation. These properties would be advantageous in treating IH. We used adenoviral mediated iNOS gene transfer in animal models of IH and showed dramatic reductions in IH using very low viral titers. The predominant actions of NO appear to be directed at SMC antiproliferation and EC growth. The efficacy of iNOS gene transfer has fueled an effort to develop this therapy for clinical applications. In this proposal, we will pursue three interrelated aims to further evaluate iNOS gene transfer. AIM 1: To determine the optimal conditions for therapeutic NOS gene transfer for the prevention of vascular injury induced IH. We will evaluate the impact of arginine and cofactor limitations on the efficacy of iNOS gene transfer. In addition, we will perform direct comparisons between the different NOS isoforms to see if any is NOS more efficacious. We will evaluate modified adenoviral vectors that target vascular cells for improving NOS gene delivery. AIM II: To evaluate the consequences of NOS gene therapy in a pi,, model of atherosclerosis. Human vascular disease occurs in the setting of atherosclerosis. Therefore, this AIM will focus on studying the impact of hypercholesterolemia and atherosclerosis on iNOS gene transfer efficiency and efficacy. We will also determine if a longer duration of NO production will be required in the setting of atherosclerosis to inhibit IH by a recombinant adeno-associated virus. AIM III: To further characterize the molecular mechanisms of NO- mediated vasoprotection. In this AIM, we will characterize the mechanisms by which NO inhibits SMC proliferation and promotes EC growth, focusing on the role of G-proteins, mitogen-activated protein kinase pathways, and cyclooxygenase-2. At the completion of our studies, we will have the necessary information to develop NOS gene therapy as a safe and effective treatment for vascular IH.

动脉粥样硬化血管疾病是美国发病率和死亡率的主要原因。诸如血管成形术和旁路程序之类的常规治疗方法因内膜增生（IH）的发展而成功限制。在过去的十年中，已经开发了基因治疗方法。我们建议预防IH的最佳候选基因之一是诱导型一氧化氮合酶（INOS）基因。 INOS会产生可以抑制平滑肌细胞（SMC）增殖，促进内皮细胞（EC）再生，抑制血小板和白细胞粘附并促进血管舒张的NO。这些特性在治疗IH方面将是有利的。我们在IH动物模型中使用了腺病毒介导的iNOS基因转移，并使用非常低的病毒滴度显示IH的急剧减少。 NO的主要作用似乎是针对SMC抗增殖和EC增长的。 iNOS基因转移的功效激发了为临床应用开发这种疗法的努力。在此提案中，我们将追求三个相互关联的目标，以进一步评估INOS基因转移。目标1：确定用于预防血管损伤诱导的IH的治疗性NOS基因转移的最佳条件。我们将评估精氨酸和辅因子局限性对iNOS基因转移功效的影响。此外，我们将在不同的NOS同工型之间进行直接比较，以查看是否更有效。我们将评估靶向血管细胞的改良腺病毒载体，以改善NOS基因递送。 AIM II：评估动脉粥样硬化模型中NOS基因疗法的后果。人血管疾病发生在动脉粥样硬化的情况下。因此，该目标将着重研究高胆固醇血症和动脉粥样硬化对iNOS基因转移效率和疗效的影响。我们还将确定在动脉粥样硬化的情况下是否需要更长的持续时间，以抑制与重组腺相关的病毒抑制IH。 AIM III：进一步表征无介导的血管保护的分子机制。在此目标中，我们将表征NO抑制SMC增殖并促进EC生长的机制，重点是G蛋白，有丝分裂原激活的蛋白激酶途径和环氧酶-2的作用。研究完成后，我们将拥有必要的信息来开发NOS基因疗法，以作为血管IH的安全有效治疗。