GENE THERAPY FOR SERUM PROTEIN DEFICIENCIES

许多重要的人类疾病是由遗传性或 后天性单一血清蛋白缺乏症。其中包括糖尿病。 糖尿病、垂体性侏儒症、血友病A、B与促红细胞生成素 (促红细胞生成素)反应性贫血。这些疾病中的许多目前都在治疗 通过反复静脉注射或皮下注射纯化的或 重组蛋白。这种疗法虽然通常是挽救生命的，但 既不方便又昂贵。此外，在某些情况下， 重组蛋白可用于所有患者的预防性治疗 而在另一些情况下，间歇性给药重组蛋白失败 以减轻这种疾病的所有副作用。这些问题 激发了人们对开发新的基因转移的兴趣 可用于治疗获得性和非获得性心脏病的治疗方法 遗传性血清蛋白缺乏。这些研究的长期目标是 在这一持续应用中描述的是肌肉的发育 一种可用于稳定编程的体细胞基因转移方法 血液中促红细胞生成素的生理调节治疗水平 患有促红细胞生成素反应性贫血的患者。要使用的方法是基于 根据这项拨款的头三年的调查结果， 证明了转基因骨骼肌细胞可以用于 稳定地将生理水平的促红细胞生成素输送到体循环 老鼠和猴子。在下面描述的研究中，调查人员 建议1)开发新型腺病毒、腺相关病毒和 促红细胞生成素基因在体骨组织转移的载体构建 肌肉；2)检测这些载体的稳定输送能力 促红细胞生成素生理水平对小鼠和猴子循环的影响 肌注后；3)发展四环素可调节和骨骼 肌肉特异性促红细胞生成素表达载体的构建及其体外和体内检测 以及4)检验低氧诱导转录的假说 EPO、LDH-A和PGK基因的调节元件可用于 构建低氧可诱导的EPO表达载体 肌注后将生理调节的促红细胞生成素输送到循环 注射。综上所述，这些研究应该为 EPO反应性患者成功的人类基因治疗试验 贫血。它们还应该与治疗一些 其他后天和遗传性血清蛋白缺乏症。

A number of important human diseases are caused by the inherited or acquired deficiency of a single serum protein. These include diabetes mellitus, pituitary dwarfism, hemophilias A an B and the erythropoietin (Epo) responsive anemias. Many of these diseases are currently treated by repeated intravenous or subcutaneous injections of purified or recombinant proteins. Such therapies although often life-saving are inconvenient and expensive. Moreover in some cases there is insufficient recombinant protein available to treat all patients prophylactically while in others intermittent administration of recombinant protein fails to ameliorate all of the side effects of the disease. These problems have stimulated interest in the development of novel gene transfer approaches that could be used for the treatment of acquired and inherited serum protein deficiencies. The long term goal of the studies described in this continuing application is the development of a muscle based somatic gene transfer approach that can be used to stably program physiologically regulated therapeutic levels of Epo in the circulation of patients with Epo responsive anemias. The method to be used is based on findings from the first three years of this grant which have demonstrated that genetically modified skeletal myocytes can be used to stably deliver physiological levels of Epo to the systemic circulation of mice and monkeys. In the studies described below the investigators propose to 1) develop novel adenoviral, adeno-associated viral and plasmid vectors for in vivo gene transfer of the Epo cDNA into skeletal muscle; 2) test the ability of these vectors to stably deliver physiological levels of Epo to the circulation of mice and monkeys following IM injection; 3) develop tetracycline-regulatable and skeletal muscle specific Epo expression vectors and test them in vitro and in vivo; and 4) test the hypothesis that hypoxia-inducible transcriptional regulatory elements from the Epo, LDH-A, and PGK genes can be used to construct hypoxia inducible Epo expression vectors that can be used to deliver physiologically regulated Epo to the circulation following IM injection. Taken together these studies should lay the foundation for successful human gene therapy trials for patients with Epo responsive anemias. They should also be relevant to the treatment of a number of other acquired and inherited serum protein deficiencies.