Generating HIV resistant cells from conditionally transformed HSC cell lines

从条件转化的 HSC 细胞系中产生 HIV 抗性细胞

• 批准号: 7419068
• 负责人: YOSEF REFAELI
• 金额: $ 16.12万
• 依托单位: TAIGA BIOTECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-19 至 2010-09-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7419068
• 关键词: AIDS therapy; Adult; Animals; Arts; Biological Assay; Blood; Blood Cells; Bone Marrow; Bone Marrow Stem Cell; Bone Marrow Stem Cell Transplantation; Bone Marrow Transplantation; CCR5 gene; CXCR4 gene; Cell Line; Cell Lineage; Cell Transplantation; Cells; Chemokine (C-C Motif) Receptor 5; Child; Clinical; Clinical Trials; Data; Engraftment; Generations; Goals; HIV; HIV-1; Hematopoietic stem cells; Human; Immune; Immune system; Immunologic Deficiency Syndromes; In Vitro; Individual; Infection; Laboratories; Leukapheresis; Lymphocyte; Malignant Neoplasms; Marrow; Methods; Methylcellulose; Modeling; Monitor; Mus; Patients; Phase; Phase I Clinical Trials; Primates; Production; Property; Public Health; Radiation; Receptor Gene; Research; Resistance; Resources; Source; Standards of Weights and Measures; Stem cell transplant; Stem cells; T-Lymphocyte; Technology; Testing; Therapeutic; Transduction Gene; Transplantation; Umbilical Cord Blood; Validation; Virus; Virus Diseases; Work; base; cell type; chemotherapy; clinical application; day; drug development; graft vs host disease; graft vs host reaction; human stem cells; in vivo; macrophage; mouse model; new technology; peripheral blood; pluripotency; receptor; reconstitution; small hairpin RNA; tool; vector

DESCRIPTION (provided by applicant): Using a method of retroviral gene transduction we have been able to generate conditionally immortalized long-term hematopoietic stem cell lines from primary mouse bone marrow stem cells. Phase I of this project includes testing the same technology in human cells and the subsequent analysis of those cells in the laboratory and in mice. In addition, the project includes knockdown of HIV-1 co-receptors to generate HIV-1 resistant cells in a mouse model. The method to genetically modify the long-term hematopoietic stem cell lines includes state-of-the-art vector transduction with shRNA containing constructs. Phase II of the project will entail testing this approach in a primate model and optimizing large-scale production of genetically-modified conditionally-immortalized long-term hematopoietic stem cells for testing in clinical trials. This project seeks to refine appropriate methods to genetically manipulate conditionally immortalized hematopoietic stem cell lines to generate HIV-1 resistant cells. Once these methods have been established for the human, the long-term goal to benefit public health will be to use these cells for treating HIV-1 infected patients to re-establish a functional immune system that is protected from endogenous or exogenous sources of HIV.

描述（由申请人提供）：使用逆转录病毒基因转导方法，我们已经能够从原代小鼠骨髓干细胞产生条件永生化长期造血干细胞系。该项目的第一阶段包括在人类细胞中测试相同的技术，以及随后在实验室和小鼠中对这些细胞进行分析。此外，该项目还包括在小鼠模型中敲低HIV-1共受体以产生HIV-1抗性细胞。遗传修饰长期造血干细胞系的方法包括用含有shRNA的构建体进行最先进的载体转导。该项目的第二阶段将需要在灵长类动物模型中测试这种方法，并优化大规模生产基因修饰的条件永生化长期造血干细胞，用于临床试验。 该项目旨在改进适当的方法，以遗传操作条件永生化造血干细胞系，以产生抗HIV-1的细胞。一旦这些方法已经为人类建立，有益于公共卫生的长期目标将是使用这些细胞来治疗HIV-1感染的患者，以重新建立功能性免疫系统，该免疫系统被保护免受内源性或外源性HIV来源的影响。