Humanized Rag2-/-gammac-/-mice for HIV infection and gene therapy

用于 HIV 感染和基因治疗的人源化 Rag2-/-gammac-/-小鼠

基本信息

批准号：
7876695
负责人：
Ramesh Akkina
金额：
$ 35.69万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-06-01 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7876695
关键词：
AIDS/HIV problem Animal Model Antibodies Antigens Antiviral Agents B-Lymphocytes CD34 gene CD4 Positive T Lymphocytes CXCR4 gene Cell surface Cells Competence Dendritic Cells Dimensions Engraftment Evaluation Gene-Modified Generations Genes HIV HIV Infections HIV-1 Hematopoiesis Hematopoietic Hematopoietic System Hematopoietic stem cells Human Immune Immune response Immune system Immunocompetent In Vitro Individual Injection of therapeutic agent Interleukin 2 Receptor Gamma Knock-out Lentivirus Vector Macaca mulatta Modeling Mus Neonatal Pathogenesis Physiological Population Predisposition Production RNA Research Resistance SCID Mice SCID-hu Mice Stem cells System T-Lymphocyte Testing Tissues Transgenic Organisms Transplantation Viral Xenograft procedure base gene therapy in vivo in vivo Model intrahepatic macrophage mouse model novel novel therapeutics preclinical evaluation reconstitution success therapeutic gene transgene expression

项目摘要

DESCRIPTION (provided by applicant): Studies on HIV pathogenesis and preclinical evaluation of HIV-1 gene therapeutic constructs would be greatly facilitated by the use of a suitable animal model that can mimic the physiological conditions afforded by a normally functioning human hematopoietic system. As the previously used xenograft SCID-hu mouse models lacked the capacity for generating primary immune responses, viral pathogenic effects on the human cells could only be evaluated in the context of the absence of an active immune response. Thus an important physiological dimension was missing in these systems. This deficiency can now be potentially rectified by the use of a newly emerged immunocompetent humanized mouse model. Injection of human CD34 hematopoietic stem cells into immunodeficient neonatal Rag2-/-yc-/- mice results in multi-lineage human hematopoiesis and the generation of a functional immune system. Recent evidence showed that these humanized mice are permissive for disseminated HIV infection. With the capacity for multi-lineage human cell engraftment and susceptibility for HIV infection, this model shows great potential to evaluate anti-HIV gene therapeutic constructs in a stem cell-based setting. Gene modified cells can now be tested for their capacity for immune reconstitution. We recently made progress in the construction and evaluation of several promising anti-HIV RNA inhibitory molecules such as siRNAs and propose to utilize this new mouse model to answer several important questions relevant for the success of gene therapy approaches. Our specific aims are to: 1. Further develop the humanized Rag2 common gamma chain double knock out (Rag2-/-yc-/-) mouse model for HIV infection to evaluate anti-HIV gene therapeutic constructs, 2. Evaluate the engraftment potential and multi-lineage differentiation of anti-HIV gene transduced CD34 hematopoietic progenitor cells in humanized Rag2-/-yc-/- mice, 3. Determine the expression of anti-HIV genes in terminally differentiated T cells and macrophages and evaluate their resistance to HIV-1 challenge in vitro and in vivo. 4. Evaluate the functional competence of transgenic hematopoietic cell subsets and the capacity of humanized Rag2-/-yc-/- mice bearing transgenic cells to give rise to normal immune responses: These studies will be helpful in developing new animal models for HIV/AIDS research, and in evaluating novel gene therapy approaches to control HIV infection.

描述（由申请人提供）：关于HIV-1基因治疗构建体的HIV发病机理和临床前评估的研究，可以通过使用合适的动物模型来大大促进，该模型可以模仿正常运作的人类造血系统所提供的生理条件。由于先前使用的异种移植SCID-HU小鼠模型缺乏产生一级免疫反应的能力，因此只有在没有主动免疫反应的背景下，才能评估对人类细胞的病毒致病作用。因此，在这些系统中缺少重要的生理维度。现在，通过使用新出现的免疫功能人性化小鼠模型可以纠正这种缺陷。将人CD34造血干细胞注射到免疫缺陷的新生儿RAG2 - / - YC - / - 小鼠会导致多乳腺癌的人类造血和功能免疫系统的产生。最近的证据表明，这些人源性小鼠允许传播HIV感染。该模型具有多种ea的人类细胞植入能力和HIV感染的易感性，在基于干细胞的环境中评估抗HIV基因治疗构建体的巨大潜力。现在，可以测试基因修饰细胞的免疫重建能力。我们最近在建造和评估几种有希望的抗HIV RNA抑制分子（例如siRNAS）方面取得了进展，并建议利用这种新的小鼠模型回答与基因治疗方法成功有关的几个重要问题。我们的具体目的是：1。进一步开发用于HIV感染的人源化RAG2常见伽马链双重敲除（RAG2 - / - YC - / - ）用于评估抗HIV基因治疗构建体的模型，2。评估植入型的抗HIV基因治疗构建体。小鼠，3。确定在终末分化的T细胞和巨噬细胞中抗HIV基因的表达，并评估其在体外和体内对HIV-1挑战的抗药性。 4。评估转基因造血细胞亚群的功能能力以及人源化RAG2 - / - YC - / - 具有转基因细胞的小鼠可导致正常的免疫反应：这些研究将有助于开发HIV/AIDS研究的新动物模型，以及评估新型基因疗法控制HIV HIV感染的方法。