GENE THERAPY OF HYPERCHOLESTEROLEMIA

这些研究的目标是开发有效的方法， 补充LDL受体和apoE基因的遗传缺陷， 家族性高胆固醇血症的基因治疗方法 我们的研究 是基于使用新型高滴度逆转录病毒载体 用水泡性口炎病毒（VSV-G）的G蛋白假型化 最近在我们的实验室里开发的。 我们的第一个主要目标是发展 更有效的方法来产生伪型载体， 主要是在开发稳定的包装细胞系， 毒性G蛋白。 利用这些向量，我们将追踪 直接体内模型，用于纠正肝脏中的遗传缺陷， LDL受体和apoE缺乏的高胆固醇血症小鼠。 的 这种方法的潜力已经被我们最近的研究所证实， 证明高效感染（>40%转导效率） 新生小鼠肝脏中的肝细胞的假型载体。 我们 还将寻求基于移植物植入的离体模型， 基因修饰的肝细胞包埋在网状聚氨酯中。 最后，我们将研究插入突变的潜力， 用假型化载体的高滴度制备物感染的细胞。

The goal of these studies is to develop efficient methods for complementing the genetic defects in the LDL receptor and apoE genes as an approach to gene therapy for familial hypercholesterolemia. Our studies are based on the use of the new class of high titer retroviral vectors pseudotyped with the G protein of vesicular stomatitis virus (VSV-G) recently developed in our laboratory. Our first major goal is to develop much more efficient methods of producing the pseudotyped vectors, aiming principally at the development of stable packaging cell lines that express the toxic G protein conditionally. Using these vectors, we will pursue direct in vivo models for correcting the genetic defects in the liver of hypercholesterolemic mice with LDL receptor and apoE deficiency. The potential for this approach has been established by our recent demonstration of highly efficient infection (>40% transduction efficiency) of hepatocytes in the neonatal mouse liver by a pseudotyped vector. We will also pursue an ex vivo model based on the implantation of grafts of genetically modified hepatocytes embedded in reticulated polyurethane. Finally, we will examine the potential for insertional mutagenesis in cells infected with high titer preparations of the pseudotyped vectors.