PATHOBIOLOGY & GENE TRANSFER IN CARDIOVASCULAR DISEASE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 家族性高胆固醇血症是一种遗传性疾病，被认为是一个很好的候选基因治疗。 在我们的家系性高胆固醇血症恒河猴中，导致低密度脂蛋白（LDL）受体缺乏的遗传缺陷已被确定为LDL受体基因外显子6的无义突变。 LDL受体基因中的这种缺陷导致在对应于人LDL受体的氨基酸284的位置处截短的蛋白质的表达。 该缺陷与自发性高胆固醇血症的表型分离了三代。 这是家族性高胆固醇血症（FH）的唯一灵长类动物模型，并且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这种疾病的重大成就。 该项目的目的是研究恒河猴LDL受体和VLDL受体基因的肝转移对LDL受体缺陷恒河猴的影响。 目的是证明转基因降低血浆LDL水平，减缓动脉病变的发展，并且在24个月内是安全的。 辅助来源的腺病毒/转基因复合物（HD-Ad-LDL-R）在小鼠中提供了超过6个月的良好表达，并且我们预期在恒河猴中得到类似的结果。 Ad-Ad在基因治疗方面有很大的前景，因为编码病毒蛋白质的基因（抗原性的来源）已经被去除。 我们第一次尝试用HD-Ad-LDL-R治疗恒河猴导致良好的基因表达和胆固醇水平降低，但仅持续2周;少于1%的辅助病毒制剂污染诱导了限制表达的免疫应答。 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.