PRECLINICAL STUDIES OF LIVER DIRECTED GENE THERAPY IN NON HUMAN PRIMATES

为了建立肝脏导向基因的临床前模型 在非人类灵长类动物的治疗中，我们比较了两种不同的疗效 载体、两种传递方式、两种报告基因和 8只恒河猴的免疫抑制。我们给两只猴子接种了 以LacZ基因为报告基因的腺病毒载体和 将它们与另外两只接种了含有LacZ的 脂质体中的质粒。每组两人通过一种 肠系膜静脉2例，经胆管2例。腺病毒导致了 比脂质体载体和肠系膜更多的基因表达 静脉接种远优于胆道接种。这个 猴子对腺病毒载体产生了免疫反应， 导致肝炎和转基因表达在2小时内丢失 几周。我们用类固醇和环磷酰胺对两只猴子进行免疫抑制 通过隐静脉将腺病毒载体1接种给他们 1例经肠系膜静脉。这两条路线都同样有效，而且 转基因表达时间延长，无肝炎发生。我们是下一个 免疫抑制2只猴子仅2周，并通过 携带含有LacZ或LacZ的腺病毒的大隐静脉 荧光素酶作为报告基因。肝炎与转基因缺失 一旦免疫抑制药物出现，就会出现表达 停产。这两种报告基因都很容易被检测到。我们计划 用更高级的载体和更精细的方法继续这些研究 免疫抑制的方法，以期建立良好的动物模型 基因治疗产品的临床前测试。

In order to develop a preclinical model of liver-directed gene therapy in nonhuman primates, we compared the efficacy of 2 different vectors, 2 modes of delivery, 2 reporter genes and of immunosuppression in 8 rhesus monkeys. We inoculated 2 monkeys with an adenovirus vector containing the lacZ gene as a reporter and compared these with 2 other monkeys inoculated with a lacZ-containing plasmid in liposomes. Two from each group were inoculated via a mesenteric vein and 2 via the bile duct. The adenovirus resulted in much more gene expression than the liposome vector, and mesenteric vein inoculation was far superior to bile duct administration. The monkeys developed an immune response to the adenoviral vector, resulting in hepatitis and loss of transgene expression within 2 weeks. We immunosuppressed 2 monkeys with steroids and cytoxan and inoculated them with the adenoviral vector, 1 via the saphenous vein and 1 via a mesenteric vein. Both routes were equally effective, and there was prolonged transgene expression and no hepatitis. We next immunosuppressed 2 monkeys for only 2 weeks and inoculated them via the saphenous vein with adenovirus containing either lacZ or luciferase as a reporter gene. Hepatitis and loss of transgene expression occurred as soon as the immunosuppressive drugs were discontinued. Both reporter genes were easily detected. We plan to continue these studies with more advanced vectors and more refined methods of immunosuppression in order to develop a good model for preclinical testing of products for gene therapy.