Generation of an HIV-1-Resistant Immune System with CD34 + Hematopoietic Stem Cells Transduced with a Triple-Combination Anti-HIV Lentiviral Vector

• DOI: 10.1128/jvi.06300-11
• Publication Date: 2012-03-08T02:57:44Z
• Authors: Jon E. Walker; Rachel X. Chen; Jeannine McGee; Catherine Nacey; Richard B. Pollard; Mehrdad Abedi; Gerhard Bauer; Jan A. Nolta; Joseph S. Anderson
• Keywords: 0301 basic medicine; Genetic Vectors; Lentivirus; Cell- and Tissue-Based Therapy; Hematopoietic Stem Cell Transplantation; Antigens, CD34; HIV Infections; Genetic Therapy; Hematopoietic Stem Cells; 3. Good health; Mice; 03 medical and health sciences; Transduction, Genetic; HIV-1; Animals; Humans

ABSTRACT

HIV gene therapy has the potential to offer an alternative to the use of current small-molecule antiretroviral drugs as a treatment strategy for HIV-infected individuals. Therapies designed to administer HIV-resistant stem cells to an infected patient may also provide a functional cure, as observed in a bone marrow transplant performed with hematopoietic stem cells (HSCs) homozygous for the CCR5-Δ32-bp allele. In our current studies, preclinical evaluation of a combination anti-HIV lentiviral vector was performed, in vivo , in humanized NOD-RAG1 −/− IL2rγ −/− knockout mice. This combination vector, which displays strong preintegration inhibition of HIV-1 infection in vitro , contains a human/rhesus macaque TRIM5α isoform, a CCR5 short hairpin RNA (shRNA), and a TAR decoy. Multilineage hematopoiesis from anti-HIV lentiviral vector-transduced human CD34 + HSCs was observed in the peripheral blood and in various lymphoid organs, including the thymus, spleen, and bone marrow, of engrafted mice. Anti-HIV vector-transduced CD34 + cells displayed normal development of immune cells, including T cells, B cells, and macrophages. The anti-HIV vector-transduced cells also displayed knockdown of cell surface CCR5 due to the expression of the CCR5 shRNA. After in vivo challenge with either an R5-tropic BaL-1 or X4-tropic NL4-3 strain of HIV-1, maintenance of human CD4 + cell levels and a selective survival advantage of anti-HIV gene-modified cells were observed in engrafted mice. The data provided from our study confirm the safety and efficacy of this combination anti-HIV lentiviral vector in a hematopoietic stem cell gene therapy setting for HIV and validates its potential application in future clinical trials.