Generating HIV resistant cells from conditionally transformed HSC cell lines

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 include s 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 genet ically 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 t o 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 protect ed from endogenous or exogenous sources of HIV.