PATHOBIOLOGY & GENE TRANSFER IN CARDIOVASCULAR DISEASE

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。家族性高胆固醇血症是一种遗传性疾病，被认为是基因治疗的极佳候选者。导致家族性高胆固醇血症恒河猴低密度脂蛋白受体缺陷的遗传缺陷已被鉴定为低密度脂蛋白受体基因外显子6的无义突变。低密度脂蛋白受体基因的这种缺陷导致了在与人低密度脂蛋白受体的284氨基酸对应的位置截断的蛋白质的表达。这一缺陷与自发性高胆固醇血症的表型经过三代分离。这是家族性高胆固醇血症(FH)的唯一灵长类动物模型，在该模型中证明基因治疗的有效性和安全性将是人类基因治疗该疾病的一项重大成就。该项目的目的是检测恒河猴低密度脂蛋白受体和极低密度脂蛋白受体基因的肝脏转移对低密度脂蛋白受体缺陷恒河猴的影响。目的是证明转基因可以降低血浆低密度脂蛋白水平，减缓动脉病变的发展，并且在24个月内是安全的。辅助衍生腺病毒/转基因复合体(HD-Ad-LDL-R)在小鼠身上可以很好地表达6个月以上，我们预计在猕猴身上也会有类似的结果。Ad-Ad为基因治疗带来了巨大的希望，因为编码病毒蛋白的基因已经被移除，病毒蛋白是抗原性的来源。我们第一次尝试用HD-Ad-LDL-R治疗恒河猴，结果是良好的基因表达和胆固醇水平的降低，但只持续了两周；不到1%的辅助病毒污染诱导了限制表达的免疫反应。该项目的结果将用于开发人体临床试验程序。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Familial hypercholesterolemia is a heritable disease that is believed to be an excellent candidate for gene therapy. The genetic defect responsible for low density lipoprotein (LDL) receptor deficiency in our pedigreed familial hypercholesterolemic rhesus monkeys has been identified as a nonsense mutation in exon 6 of the LDL receptor gene. This defect in the LDL receptor gene results in the expression of a protein truncated at a position corresponding to amino acid 284 of the human LDL receptor. The defect has segregated with the phenotype of spontaneous hypercholesterolemia through three generations. This is the only primate model of familial hypercholesterolemia (FH), and proof of efficacy and safety of gene therapy in this model would be a major accomplishment toward human gene therapy for this disease. The objective of the program project is to examine the effect of the hepatic transfer of rhesus LDL receptor and VLDL receptor genes to LDL receptor defective rhesus monkeys. The goal is to demonstrate that transgenes reduce plasma LDL levels, slow development of arterial lesions, and are safe during a 24-month period. The helper-derived adenovirus/transgene complex (HD-Ad-LDL-R) provides good expression in mice for more than 6 months and we anticipated similar results in rhesus. Ad-Ad holds great promise for gene therapy since the genes coding for viral proteins, the source of antigenicity, have been removed. Our first attempts to treat rhesus with HD-Ad-LDL-R resulted in good gene expression and reduction in cholesterol levels, but for only 2 weeks; less than 1% contamination of the preparation with helper virus induced an immune response that limited expression. The results of this project will be used to develop procedures for human clinical trials.