MODELLING GENE THERAPEUTICS APPROACHES IN SCID-HU MICE

人类基因疗法正被积极考虑作为一种可能的治疗方法 针对感染人类免疫缺陷病毒1型（HIV-1）个人的战略 感染或艾滋病。有许多基因治疗策略 提出，包括反式显性蛋白，核酶和反义RNA， 在实验室中已经成功地用于阻断 HIV-1在培养中的复制。我们已经开发了严重的组合 免疫缺陷（SCID）小鼠作为HIV-1感染和 发病机制和作为一个实验系统，其中研究基因 治疗方法。SCID小鼠可以移植人类胸腺 （SCID-hu），其在CD 34+祖细胞的存在下，将产生 使CD 4+和CD 8 + T细胞成熟。我们已经证明，感染这些 携带HIV-1的人体移植会在三到四年内导致病理学改变， 我们最近已经证明，干细胞可以被转导， 与适合于基因治疗目的的载体一起使用，使得成熟的T- 细胞在分化后也携带载体。虽然这 模型系统不能完全代表正常人造血 和/或HIV-1感染的所有方面，HIV-1病理学的组合 和CD 34+祖细胞转导发生在相对较短的时间内， 短时间内为我们提供了一个实验系统， 解决许多关键问题，成功的基因治疗方法 HIV-1和艾滋病。 该应用程序构成两部分交互式RO 1的一部分， 旨在为HIV-1疾病的基因治疗方法建模， SCID小鼠。所附申请题为“免疫”， 在SCID-hu小鼠中的重建和基因转导”（杰罗姆Zack， 主要研究者）组成了该交互式RO 1的第二部分。 该提案旨在进一步开发SCID-hu小鼠， 了解干细胞重建所需的因素， 在SCID小鼠中的分化。本提案侧重于矢量测试 并使用SCID-hu小鼠模拟特定的基因治疗方法。 该提案的具体目标是： 1.逆转录病毒载体转导CD 34+细胞的参数优化 干细胞和重建在SCID小鼠。 2.腺相关病毒载体转导参数的优化 转化为CD 34+干细胞并在SCID小鼠中重建。 3.检测利用SCID-hu小鼠模型进行检测的可行性 选择基因治疗方法。 因为没有已知的基因疗法 我们将选择两种基因疗法 建模方法：a）转显性;和B）基于RNA的。 我们将 确定各种抗逆转录病毒基因对HIV-1的疗效- 根据目标3的结果，诱导病理学。

Human gene therapy is being actively considered as a possible treatment strategy for individuals with human immunodeficiency virus type 1 (HIV-1) infection or AIDS. There have been numerous gene therapeutic strategies proposed, including transdominant proteins, ribozymes and anti-sense RNAs, which have been utilized with variable success in the laboratory to block HIV-1 replication in culture. We have developed the severe combined immunodeficient (SCID) mouse as a model for both HIV-1 infection and pathogenesis and as an experimental system in which to investigate gene therapy approaches. The SCID mice can be transplanted with a human thymus (SCID-hu) which, in the presence of CD34+ progenitor cells, will give rise to mature CD4 + and CD8 + T-cells. We have shown that infection of these human transplants with HIV-1 results in pathology within three to four weeks, and we have shown more recently that stem cells can be transduced with vectors suitable for gene therapy purposes such that the mature T- cells also harbor the vector following differentiation. Although this model system is not perfectly representative of normal human hematopoiesis and/ or all aspects of HIV-1 infection, the combination of HIV-1 pathology and CD34+ progenitor cell transduction taking place within a relatively short period of time provides us with an experimental system in which to address many of the issues critical to successful gene therapy approaches for HIV-1 and AIDS. This application constitutes one part of a two-part interactive RO1 which aims to model gene therapy approaches for HIV-1 disease, utilizing the SCID-hu mouse. The accompanying application entitled, "Immune Reconstitution and Gene Transduction in the SCID-hu Mouse" (Jerome Zack, Principal Investigator) comprises the second part of this interactive RO1. That proposal aims to further develop the SCID-hu mouse and better understand the factors required for stem cell reconstitution and differentiation in the SCID mouse. This proposal focuses on vector testing and modelling specific gene therapy approaches using the SCID-hu mouse. The Specific Aims of this proposal are: 1. Optimize parameters for transduction of retroviral vectors into CD34+ stem cells and reconstitution in the SCID mouse. 2. Optimize parameters for transduction of adeno-associated virus vectors into CD34+ stem cells and reconstitution in the SCID mouse. 3. Test the feasibility of utilizing the SCID-hu mouse model to test selected gene therapy approaches. Since there are no known gene therapies that are currently successful for HIV-1, we will select two gene therapy approaches to model: a) transdominant; and b) RNA-based. We will determine the efficacy of various antiretroviral genes against HIV-1- induced pathology, based upon the results of Aim 3.