Genetic modification of hematopoietic stem cells (HSC) the potential of reconstituting immune function in HIV-infected with a lifelong source of hematopoietic cells resistant to HIV infection. However, retroviral vectors based on Moloney murine leukemia virus have proved to be relatively inefficient in transducing primate HSC, in large part due to the inability of these vectors to transduce quiescent cells. Construction of retroviral vectors based on lentiviral vectors has resulted in efficient transduction of many quiescent cell types, including hematopoietic progenitor cells, and suggests that these vectors may efficiently transduce HSC. However, demonstration of the efficacy and safety of these vectors for HSC gene therapy will require in vivo studies. Recent collaborative work between our laboratory and Dr. Wong-Staal's laboratory has demonstrated that introduction of an SIV-specific ribozyme into rhesus CD34+ hematopoietic cells can induce a relatively high level of resistance to SIV infection in both CD4+ T cells and macrophages derived from these cells. The objective of this proposal is to examine the ability of HIV and FIV- based vectors encoding an SIV-specific ribozyme to transduce HSC and to protect hematopoietic progeny from SIV infection both in vitro and in vivo.
Specific aims i nclude: 1. To assess the ability of HIV and FIV vectors encoding an SIV-specific ribozyme to transduce rhesus hematopoietic progenitor cells in vitro and to inhibit SIV replication in their progeny; 2. To examine the ability of lentivirus-based vectors to transduce HSC in normal macaques; and 3. To examine the ability of HSC transduced with lentivirus vectors encoding an SIV-specific ribozyme to protect hematopoietic cells from SIV infection in vivo. 4: To determine the potential mobilization and pathogenicity of HIV and FIV vectors in rhesus macaques in vivo. These studies should yield important information regarding the efficacy and safety of lentivirus vectors for stem cell gene therapy for AIDS.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
3P01AI045992-01S1
Application #
6314015
Study Section
Special Emphasis Panel (ZAI1)
Project Start
1999-08-15
Project End
2000-07-31
Budget Start
Budget End
Support Year
1
Fiscal Year
2000
Total Cost
$208,104
Indirect Cost
Name
University of California San Diego
Department
Type
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Cheng, Lingyun; Toyoguchi, Mitsuko; Looney, David J et al. (2005) Efficient gene transfer to retinal pigment epithelium cells with long-term expression. Retina 25:193-201
Morris, Kevin V; Looney, David J (2005) Characterization of human immunodeficiency virus (HIV)-2 vector mobilization by HIV-1. Hum Gene Ther 16:1463-72
Morris, Kevin V; Chan, Simon W-L; Jacobsen, Steven E et al. (2004) Small interfering RNA-induced transcriptional gene silencing in human cells. Science 305:1289-92
Gruber, A; Chen, I; Kuhen, K L et al. (2003) Generation of dendritic cells from lentiviral vector-transduced CD34+ cells from HIV+ donors. J Med Virol 70:183-6
Cheng, Lingyun; Chaidhawangul, Sunan; Wong-Staal, Flossie et al. (2002) Human immunodeficiency virus type 2 (HIV-2) vector-mediated in vivo gene transfer into adult rabbit retina. Curr Eye Res 24:196-201
Gruber, A; Wheat, J C; Kuhen, K L et al. (2001) Differential effects of HIV-1 protease inhibitors on dendritic cell immunophenotype and function. J Biol Chem 276:47840-3
Gruber, A; Looney, D J; Ibanez, M et al. (2001) Altered immunophenotype of dendritic cells generated from HIV infected subjects. Immunol Lett 78:209-11
Sauter, S L; Gasmi, M (2001) FIV vector systems. Somat Cell Mol Genet 26:99-129
Gruber, A; Kan-Mitchell, J; Kuhen, K L et al. (2000) Dendritic cells transduced by multiply deleted HIV-1 vectors exhibit normal phenotypes and functions and elicit an HIV-specific cytotoxic T-lymphocyte response in vitro. Blood 96:1327-33