TESTING ANTIHIV-1 GENE THERAPY IN SCID-HU MICE

本实验的目的是开发SCID-HU 以小鼠为模型系统检测针对阻断的基因疗法 HIV-1复制。基因治疗提供了相当大的优势，作为一种 开发艾滋病治疗方法的范例。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 这些都是困难、昂贵的，而且可能会产生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 系统检测抗艾滋病病毒-L基因疗法，因为它是最好的艾滋病 目前可用的发病机制模型系统。体外进化REV 亲和力高于野生型的结合元件(RBE)RNA分子- 被称为RBE适配子-将被用作艾滋病基因治疗的原型 探员。这些分子由安德鲁·艾灵顿博士提供 印第安纳大学，应该有抗HIV-1的活性，因为他们 其他人展示的RBE诱饵的更高亲和力版本 在组织培养中抑制HIV-1复制。我们将使用几种形式的 单独的或在整个REV响应的上下文中的RBE适配子 各种RNA聚合酶II或RNA聚合酶表达的元件(RRE) 逆转录病毒载体中的启动子。这些向量将用于 体外转化细胞系和正常T细胞及纯化的人T细胞 CD34+造血祖细胞。Rbe适体转化组织培养 细胞将受到非合胞体诱导(NSI)和合胞体- 诱导(SI)HIV-1分子克隆和患者分离株的初步研究 RBE适配子活性检测。Rbe适配子转化CD34+造血细胞 祖细胞将被用于重新填充人类胸腺/肝脏移植物 照射后的SCID-HU小鼠。这些移植物随后将感染NSI 和SI毒株的HIV-1来确定RBE适配子是否可以抑制 病毒复制和发病机制。其他抗HIV-1基因疗法 待SCID-HU小鼠检测体系优化后，再对试剂进行检测。 最终目标是确定哪些基因治疗策略是最有效的 可能在患者身上起作用的只有最有效的艾滋病基因 治疗药物需要在临床试验中进行测试。这一模式的使用 系统将加速寻找有效的艾滋病治疗方法的进程 一个或多个代理。

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.