MODIFIED ADENOVIRUS VECTORS FOR USE IN GENE THERAPY

腺病毒载体具有将转基因运送到多种细胞的能力 不同于基于逆转录病毒的载体，Ad. 载体还可以有效地转导有丝分裂静止的细胞。 因此，许多不同疾病的潜在治疗方法，既有 通过使用Ad载体，可以预见遗传性和非遗传性。 例如，广告载体已经被证明能够 将基因输送到肝细胞，用于潜在的治疗多发性硬化 代谢紊乱，2)肌肉细胞(骨骼和心脏) 肌病和储存障碍的潜在治疗，3)大脑和 神经系统组织在神经系统治疗中的潜在应用 疾病，如帕金森氏病，以及4)呼吸道上皮 治疗囊性纤维化等肺部疾病。此外, 许多其他常见的疾病，如艾滋病和各种形式的癌症 所有这些都被证明是由Ad介导的基因潜在地治疗的 转移策略。虽然存在巨大的潜力，但 治疗许多人类疾病，有几个问题 在Ad介导的基因之前必须解决的当前Ad载体 治疗变成了临床现实。最严重的问题是 当前的Ad载体是转基因表达的瞬时持续时间 在成功地将基因转移到免疫活性组织后 动物。其他问题包括复制能力的产生 Ad(RCA)，以及Ad载体无法携带更大的基因或 组织特异性启动子/增强子元件。这项拨款建议 概述了一系列实验，这些实验将解决每个 当前广告载体的局限性。在这样做的时候，我们将孤立 修改的广告载体，预计将允许更长的持续时间 体内转基因表达，降低RCA的发生率， 并显著提高了Ad载体的载量。最初， 改良的Ad载体将在小鼠肝脏和肌肉模型中进行分析 细胞基因疗法。其结果将是分离出新的广告载体 也能够在人类疾病的动物模型中有效使用 至于最终用于治疗大量人类 条件。

Ad vectors have the ability to deliver transgenes to a variety of cell types, in vitro and in vivo, and unlike retrovirus based vectors, Ad vectors can also efficiently transduce mitotically quiescent cells. Therefore, the potential treatment of many different diseases, both genetic and non-genetic can be envisioned with the use of Ad vectors. For example, Ad vectors have been demonstrated to be capable of delivering genes to 1) liver cells for the potential treatment of many metabolic disorders, 2) muscle cells (skeletal and cardiac) for the potential treatment of myopathies and storage disorders, 3) brain and nervous system tissues for the potential treatment of neurologic diseases like Parkinson disease, and 4) respiratory epithelium for the treatment of pulmonary disorders like cystic fibrosis. In addition, many other common diseases like AIDS and various forms of cancer have all been demonstrated to be potentially treated by Ad mediated gene transfer strategies. While there is an enormous potential for the treatment of many human diseases, there are several problems with current Ad vectors that must be addressed before Ad mediated gene therapy becomes a clinical reality. The most serious problem with current Ad vectors is the transient duration of transgene expression after successful gene delivery into the tissues of immunocompetent animals. Other problems include the generation of replication competent Ad (RCA), and the inability of Ad vectors to carry larger genes or tissue-specific promoter/enhancer elements. This grant proposal outlines a series of experiments that will address each of the limitations of current Ad vectors. In so doing, we will isolate modified Ad vectors that are predicted to allow for longer durations of transgene expression in vivo, decrease the incidence of RCA generation, and significantly increase Ad vector carrying capacity. Initially, the modified Ad vectors will be analyzed in mouse models of liver and muscle cell gene therapy. The result will be the isolation of new Ad vectors capable of efficacious use in animal models of human disease, as well as for eventual use in the therapy of a great number of human conditions.