GENE THERAPY FOR SERUM PROTEIN DEFICIENCIES

许多重要的人类疾病是由遗传或 一种血清蛋白的获得性缺乏。 其中包括糖尿病 糖尿病、垂体性侏儒症、血友病A和B以及促红细胞生成素 (Epo)反应性贫血 这些疾病中有许多目前正在治疗 通过反复静脉内或皮下注射纯化的或 重组蛋白 这种疗法虽然常常能挽救生命， 不方便且昂贵。此外，在某些情况下， 可用于治疗所有患者的重组蛋白 而在其它情况下，间歇性施用重组蛋白失败 来减轻疾病的所有副作用。 这些问题 激发了人们对开发新型基因转移的兴趣， 可用于治疗后天性和 遗传性血清蛋白缺乏症 研究的长期目标 在该连续申请中描述了肌肉的发育 基于体细胞基因转移的方法，可用于稳定编程 循环中生理调节的Epo治疗水平 对EPO有反应的贫血患者。 使用的方法基于 根据这项赠款前三年的调查结果， 证明了基因修饰的骨骼肌细胞可用于 稳定地将生理水平的Epo递送到体循环 老鼠和猴子。 在下面描述的研究中，研究人员 建议1）开发新的腺病毒，腺相关病毒， 用于将Epo cDNA体内基因转移到骨骼肌中的质粒载体 2）测试这些载体稳定递送的能力 Epo对小鼠和猴子循环的生理水平 IM注射后; 3）发展四环素可调节和骨骼 肌肉特异性Epo表达载体，并在体外和体内测试它们。 体内;和4）测试假设，缺氧诱导的转录 来自Epo、LDH-A和PGK基因的调节元件可用于 构建低氧诱导型Epo表达载体， 在IM后将生理调节的Epo输送到循环中 注射 这些研究加在一起，应该为以下方面奠定基础： 成功的人类基因治疗试验，用于Epo反应性患者 贫血 它们还应与处理一些 其他获得性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.