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: 6189142
• 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: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6189142
• 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）的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的风险增加有关，但载脂蛋白A-I基因中米兰突变的人类携带者是值得注意的，因为尽管HDL-C水平极低，但他们享有免于血管疾病的自由，并且具有长寿的祖先史。这些观察结果表明，与野生型apo A-I、apo A-I/wt.虽然这些rHDL颗粒的静脉内给药的结果提供了它们抗动脉粥样硬化功效的证据，但需要重复静脉内给药仍然是一个实际限制。因此，体细胞基因治疗诱导在体内生产的载脂蛋白A-I提供了一种替代的方法，利用载脂蛋白A-I的抗动脉粥样硬化的作用。最近，已经证明了通过重组腺相关病毒（rAAV）载体递送的转基因的长期体内表达。因此，在本项目中，我们将探讨利用含有载脂蛋白A-I/wt和载脂蛋白A-I/Milano基因的rAAV载体在载脂蛋白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中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。在猪血管损伤模型中应用apo A-I/wt/apo A-I/Milano基因进行基因治疗，可提供证明apo A-I基因治疗在人类中的可行性所需的临床前研究类型。具体目标是：1）在体内施用编码apo A-I/wt和apo A-I/Milano基因的rAAV载体后小鼠中转基因表达的分析，2）rAAV介导的apo A-I/wt和apo A-I/Milano基因的转移对apo E缺陷小鼠中对颈动脉损伤的动脉反应的影响，3）使用用编码人apo A-I/wt和apo A-I/Milano的rAAV载体转导的巨噬细胞评估基因转移的可行性和功效，和4）评价局部rAAV介导的人apo A-I/wt和apo A-I/Milano基因转移对猪中对球囊过度拉伸和支架过度膨胀损伤的冠状动脉管腔狭窄反应的影响。