PATHOBIOLOGY & GENE TRANSFER IN CARDIOVASCULAR DISEASE

该子项目是利用 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来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 家族性高胆固醇血症（FH）是一种遗传性疾病，被认为是一个很好的候选基因治疗。 SFBR维持了一个独特的恒河猴（Macacca mulatta）群体，该群体携带低密度脂蛋白（LDL）受体的缺陷基因，并提供了一个自然发生的人类家族性高胆固醇血症模型。 这些猴的LDL受体缺陷已被鉴定为外显子6中的无义突变，其导致产生在对应于人LDL受体的氨基酸284的位置处截短的蛋白质。 这种缺陷与自发性高胆固醇血症的表型分离了四代。 这是FH的唯一非人灵长类动物模型，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这种疾病的重大成就。 该项目的目的是检查恒河猴和人LDL受体基因的肝转移对LDL受体缺陷恒河猴的影响。 目的是证明转基因增加LDL受体的肝脏表达，降低血浆LDL水平，减缓动脉病变的发展，并且在24个月期间是安全的。 三种方法用于肝递送具有LDL-R的辅助依赖性腺病毒载体（HDAd-LDLR）。 首先，静脉内（IV）给予5x 10 E12 vp/kg HDAd-LDLR的单次注射。在注射后21天，治疗将血浆胆固醇水平从578 mg/dl降低至276 mg/dl;然而，胆固醇在42天恢复至治疗前水平。治疗与肝酶升高相关。 第二，肝细胞转导的效率增加了直接HDAd注射到肝动脉，同时暂时提高肝内压的球囊导管插入下腔静脉（IVC），阻断肝静脉引流，这是释放前立即HDAd注射。

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 (FH) is a heritable disease that is believed to be an excellent candidate for gene therapy. SFBR maintains a unique colony of rhesus monkeys (Macacca mulatta) that carry a defective gene for the low density lipoprotein (LDL) receptor and provide a naturally occurring model of human familial hypercholesterolemia. The defect in LDL receptor of these monkeys has been identified as a nonsense mutation in exon 6 that results in the production of a protein truncated at a position corresponding to amino acid 284 of the human LDL receptor. This defect has segregated with the phenotype of spontaneous hypercholesterolemia through four generations. This is the only nonhuman primate model for 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 and human LDL receptor genes to LDL receptor defective rhesus monkeys. The goal is to demonstrate that transgenes increase hepatic expression of LDL receptor, reduce plasma LDL levels, slow development of arterial lesions and are safe during a 24-month period. Three methods were used for hepatic delivery of helper-dependent adenoviral vector with LDL-R (HDAd-LDLR). First, a single injection of 5 x 10E12 vp/kg HDAd-LDLR was given intravenously (IV). The treatment lowered the plasma cholesterol level from 578 mg/dl to 276 mg/dl at 21 days after injection; however, cholesterol returned to the pretreatment level by 42 days. Treatment was associated with elevated liver enzymes. Second, the efficiency of hepatocyte transduction was increased by a direct HDAd injection into the hepatic artery while transiently raising intrahepatic pressure by a balloon catheter inserted in the inferior vena cava (IVC), blocking hepatic venous drainage, which was released immediately before HDAd injection.