Generating HIV resistant cells from conditionally transformed HSC cell lines

描述（由申请人提供）：使用逆转录病毒基因转导的方法，我们已经能够从原代小鼠骨髓干细胞中产生有条件永生的长期造血干细胞系。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除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最先进的shRNA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，并优化大规模生产用于临床试验的转基因条件永生长期造血干细胞。

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.