AAV MEDIATED APO A1 GENE THERAPY FOR ATHEROSCLEROSIS

AAV 介导的 APO A1 基因治疗动脉粥样硬化

• 批准号: 6338900
• 负责人: Prediman Krishan Shah
• 金额: $ 34.71万
• 依托单位: CITY OF HOPE NATIONAL MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6338900
• 关键词: adeno associated virus group; antiatherogenic agent; apolipoprotein E; apolipoproteins; artery stenosis; atherosclerotic plaque; gene therapy; genetically modified animals; high density lipoproteins; human genetic material tag; laboratory mouse; nonhuman therapy evaluation; swine; vascular endothelium

Animal studies have provided compelling evidence in favor of direct anti- atherogenic effects pf JD;-C and apolipoprotein A I (apo A-I). While low HDL-C levels are often associated with an increased risk of coronary artery diseases, human carriers of the Milano mutation in the apo A-I gene are remarkable in that despite extremely low levels of HDL-C, they enjoy freedom from vascular disease and have an ancestral history of longevity. These observations have suggested that the apo A-I/Milano, compared to the wild type apo A-I, apo A-I/wt. While the results of intravenous administration of these rHDL particles have provided evidence of their anti- atherogenic efficacy, the need for repeated intravenous administration remains a practical limitation. Therefore, somatic gene therapy to induce in vivo production of apo A-I offers an alternative approach for utilizing the anti-atherogenic effects of apo A-I. Recently, long-term in vivo expression of transgenes delivered by recombinant adeno-associated virus (rAAV) vectors have been demonstrated. Therefore, in this project we will explore the feasibility and biologic efficacy of gene therapy utilizing rAAV vectors containing the apo A-I/wt and apo A-I/Milano genes in preventing atherosclerosis and pot-injury intimal hyperplasia in the apo E-deficient mouse model. Studies using mouse model of arterial injury will be extended to a large animal model to examine the effects of local gene delivery on the coronary arterial luminal narrowing following balloon overstretch and stent over-inflation injury in swine. The application of gene therapy with apo A- I/wt/apo A-I/Milano gene in the swine vascular injury model may provide the type of preclinical studies needed to demonstrate the feasibility of apo A-I gene therapy in humans. The specific aims are: 1) Analysis of transgene expression in mice following in vivo administration of rAAV vectors encoding apo A-I/wt and apo A-I/Milano genes, 2) Effect of rAAV- mediated transfer of apo A-I/wt and apo A-I/Milano genes on arterial response to carotid injury in apo E-deficient mice, 3) Assessment of feasibility and efficacy of gene transfer using macrophages transduced with rAAV vectors encoding human apo A-I/wt and apo A-I/Milano, and 4) Evaluation of the effects of local rAAV mediated transfer of human apo A- I/wt and apo A-I/Milano genes on coronary arterial luminal narrowing response to balloon-overstretch and stent-overinflation injury in swine.

动物研究提供了令人信服的证据，支持 JD;-C 和载脂蛋白 A I (apo A-I) 具有直接抗动脉粥样硬化作用。虽然低 HDL-C 水平通常与冠状动脉疾病风险增加相关，但 apo A-I 基因 Milano 突变的人类携带者值得注意的是，尽管 HDL-C 水平极低，但他们没有血管疾病，并且祖传的长寿史。这些观察表明，与野生型apo A-I相比，apo A-I/Milano、apo A-I/wt。虽然这些rHDL颗粒的静脉内施用的结果已经提供了它们的抗动脉粥样硬化功效的证据，但是重复静脉内施用的需要仍然是实际的限制。因此，诱导体内产生apo A-I 的体细胞基因治疗为利用apo A-I 的抗动脉粥样硬化作用提供了另一种方法。最近，重组腺相关病毒（rAAV）载体递送的转基因的长期体内表达已得到证实。因此，在本项目中，我们将探讨利用含有apo A-I/wt和apo A-I/Milano基因的rAAV载体进行基因治疗在apo E缺陷小鼠模型中预防动脉粥样硬化和罐损伤内膜增生的可行性和生物学功效。使用小鼠动脉损伤模型的研究将扩展到大型动物模型，以检查局部基因递送对猪球囊过度拉伸和支架过度膨胀损伤后冠状动脉管腔狭窄的影响。 apo A-I/wt/apo A-I/Milano 基因的基因治疗在猪血管损伤模型中的应用可能提供所需的临床前研究类型，以证明 apo A-I 基因治疗在人类中的可行性。具体目标是：1) 体内施用编码 apo A-I/wt 和 apo A-I/Milano 基因的 rAAV 载体后分析小鼠中的转基因表达，2) rAAV 介导的 apo A-I/wt 和 apo A-I/Milano 转移的效果apo E 缺陷小鼠动脉对颈动脉损伤反应的基因，3) 使用 rAAV 转导的巨噬细胞评估基因转移的可行性和功效编码人apo A-I/wt和apo A-I/Milano的载体，和4)评估局部rAAV介导的人apo A-I/wt和apo A-I/Milano基因转移对冠状动脉管腔狭窄对球囊过度拉伸和收缩反应的影响。猪的支架过度充气损伤。