GENE THERAPY OF HYPERCHOLESTEROLEMIA

这些研究的目标是开发有效的方法来 补充低密度脂蛋白受体和载脂蛋白E基因的遗传缺陷作为一种 家族性高胆固醇血症的基因治疗探讨我们的研究 是基于新型高滴度逆转录病毒载体的使用 水疱性口炎病毒G蛋白的假型 是我们实验室最近开发出来的。我们的第一大目标是发展 产生伪类型向量的更有效的方法，瞄准 主要是发展稳定的包装细胞系，表达 有条件的有毒G蛋白。利用这些载体，我们将继续 直接活体动物模型用于纠正小鼠肝脏遗传缺陷 低密度脂蛋白受体和载脂蛋白E缺乏的高胆固醇血症小鼠。这个 这种方法的潜力已经被我们最近的 高效感染的示范(&gt；40%转导效率) 新生小鼠肝脏中肝细胞的假类型载体。我们 还将寻求一种基于移植的体外模型 转基因肝细胞包埋在网状聚亚安酯中。 最后，我们将研究插入突变的可能性。 用高滴度的伪型载体制剂感染细胞。

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.