TESTING ANTIHIV-1 GENE THERAPY IN SCID-HU MICE

本文提出的实验目的是开发SCID-hu 小鼠作为测试针对阻断的基因疗法的模型系统 HIV-1复制。 基因治疗作为一种治疗方法， 发展艾滋病疗法的范例。过多的可能目标 然而，治疗策略指出需要一个模型系统， 在临床试验之前测试潜在的艾滋病基因疗法 这是困难的，昂贵的，可能有不可预见的不利影响， 对人类受试者的影响。SCID-hu小鼠是一个很有吸引力的模型 系统测试抗HIV-1基因疗法，因为它是最好的艾滋病 发病机制模型系统目前可用。体外进化Rev 结合元件（RBE）RNA分子具有高于野生型的亲和力- 称为RBE适体-将被用作艾滋病基因治疗的原型 剂 这些分子由安德鲁·艾灵顿博士提供 印第安纳州大学的研究人员，应该具有抗HIV-1活性，因为他们是 其他人已经证明，更高亲和力的RBE诱饵， 抑制组织培养中HIV-1复制。我们将使用几种形式的 RBE适体，单独或在整个Rev应答的背景下 由各种RNA聚合酶II或RNA聚合酶表达的元件（RRE） III启动子。这些向量将用于 体外转化细胞系和正常T细胞以及纯化的人 CD 34+造血祖细胞。RBE适体转化的组织培养 细胞将用非合胞体诱导（NSI）和合胞体- 诱导（SI）HIV-1分子克隆和患者分离株作为初步的 RBE适体活性测试。RBE适体转化的CD 34+造血 祖细胞将用于重新填充人胸腺/肝脏移植物 辐射的SCID-hu小鼠。这些移植物会被NSI感染 和SI株的HIV-1，以确定RBE适体是否可以抑制 病毒复制和发病机制。其他抗HIV-1基因治疗剂 当SCID-hu小鼠测定系统优化时，将测试试剂。 最终的目标是确定哪些基因治疗策略是最重要的。 这样只有最有效的艾滋病基因 治疗方法需要在临床试验中进行测试。该模型的使用 该系统将加速寻找有效的艾滋病治疗方法的进程 代理或代理。

The purpose of the experiments proposed here is to develop the SCID-hu mouse as model system for testing gene therapeutics directed at blocking HIV-1 replication. Gene therapy offers considerable advantages as a paradigm for developing AIDS therapies. The plethora of possible targets and therapeutic strategies, however, points to the need for a model system in which to test potential AIDS gene therapies prior to clinical trials which are difficult, expensive and may have unforeseen adverse consequences for human subjects. The SCID-hu mouse is an attractive model system to test anti-HIV-l gene therapies since it is the best AIDS pathogenesis model system currently available. In vitro evolved Rev binding element (RBE) RNA molecules with higher than wild type affinity - called RBE aptamers - will be used as a prototype AIDS gene therapeutic agent. These molecules, which have been provided by Dr. Andrew Ellington of Indiana University, should have anti-HIV-1 activity since they are higher affinity versions of RBE decoys which have been shown by others to inhibit HIV-1 replication in tissue culture. We will use several forms of the RBE aptamers, alone, or in the context of the entire Rev response element (RRE) expressed from various RNA polymerase II or RNA polymerase III promoters in retroviral vectors. These vectors will be used to transform cell lines and normal T-cells in vitro as well as purified human CD34+ hemopoietic progenitor cells. RBE aptamer transformed tissue culture cells will be challenged with non syncytium-inducing (NSI) and syncytium- inducing (SI) HIV-1 molecular clones and patient isolates as a preliminary test of RBE aptamer activity. RBE aptamer transformed CD34+ hemopoietic progenitor cells will be used to repopulate the human thymus/liver grafts of irradiated SCID-hu mice. These grafts will then be infected with NSI and SI strains of HIV-1 to determine whether the RBE aptamers can inhibit viral replication and pathogenesis. Other anti-HIV-1 gene therapeutic agents will be tested when the SCID-hu mouse assay system is optimized. The ultimate goal is to define which gene therapeutic strategies are most likely to work in patients so that only the most potent AIDS gene therapeutics need be tested in clinical trials. The use of this model system will hasten the process of finding an effective AIDS therapeutic agent or agents.