MODELLING GENE THERAPEUTIC APPROACHES IN SCID/HU MICE

SCID/HU 小鼠基因治疗方法建模

• 批准号: 2413694
• 负责人: IRVIN S.Y. CHEN
• 金额: $ 28.42万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1998-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2413694
• 关键词: AIDS; CD34 molecule; CD4 molecule; CD8 molecule; SCID mouse; adeno associated virus group; biopsy; cell sorting; disease /disorder model; gene expression; gene therapy; genetic promoter element; genetic transduction; hematopoietic stem cells; human fetus tissue; human immunodeficiency virus 1; liposomes; messenger RNA; murine AIDSs; pathologic process; polymerase chain reaction; southern blotting; thymus; thymus transplantation; virus replication

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.

人类基因疗法正被积极考虑作为一种可能的治疗方法。 针对人类免疫缺陷病毒1型(HIV-1)患者的策略 感染或艾滋病。已经有许多基因治疗策略 提出的，包括跨显性蛋白、核酶和反义RNA， 它们已经在实验室中成功地被用来阻止 HIV-1在培养上的复制。我们已经开发出了严重的联合 免疫缺陷(SCID)小鼠作为HIV-1感染和 发病机制和作为研究基因的实验系统 治疗即将到来。SCID小鼠可以移植人胸腺 (SCID-Hu)，在CD34+祖细胞存在的情况下，将产生 成熟的CD4+和CD8+T细胞。我们已经证明了这些病毒的感染 携带HIV-1的人体移植在三到四个月内导致病理 几周后，我们最近证明了干细胞可以被转导 具有适合于基因治疗目的的载体，以便成熟的T- 分化后的细胞也含有该载体。虽然这件事 模型系统不能很好地代表正常的人类造血 和/或艾滋病毒-1感染的方方面面，艾滋病毒-1病理的组合 和CD34+祖细胞转导发生在相对 短时间为我们提供了一个实验系统，在其中 解决许多对成功的基因治疗方法至关重要的问题 针对HIV-1和艾滋病。 该应用程序构成由两部分组成的交互式RO1的一部分， 旨在为HIV-1疾病的基因治疗方法建模，利用 SCID-HU小鼠。随附的申请书标题为“免疫” 在SCID-HU小鼠中的重组和基因转导“(Jerome Zack， 首席调查员)构成了这一互动RO1的第二部分。 该建议旨在进一步开发SCID-HU小鼠，并更好地 了解干细胞重建所需的因素和 在SCID小鼠中的分化。本提案的重点是病媒测试 并使用SCID-HU小鼠模拟特定的基因治疗方法。 这项建议的具体目标是： 1.优化逆转录病毒载体转导CD34+的参数 干细胞与SCID小鼠的重建。 2.优化腺相关病毒载体的转导参数 转化为CD34+干细胞，并在SCID小鼠体内重建。 3.验证利用SCID-HU小鼠模型进行实验的可行性 选定的基因治疗方法。因为目前还没有已知的基因疗法 对于目前成功的HIV-1，我们将选择两种基因疗法 建模方法：a)跨显性；b)基于RNA。我们会的 确定各种抗逆转录病毒基因对HIV-1的疗效- 诱导病理，基于目标3的结果。