title: Generation of an HIV-1-resistant immune system with CD34(+) hematopoietic stem cells transduced with a triple-combination anti-HIV lentiviral vector.
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.
authors.affiliation: Department of Internal Medicine, University of California-Davis Medical Center, Sacramento, California, USA.
authors.forename: Jon E
authors.initials: JE
authors.lastname: Walker
authors.forename: Rachel X
authors.initials: RX
authors.lastname: Chen
authors.forename: Jeannine
authors.initials: J
authors.lastname: McGee
authors.forename: Catherine
authors.initials: C
authors.lastname: Nacey
authors.forename: Richard B
authors.initials: RB
authors.lastname: Pollard
authors.forename: Mehrdad
authors.initials: M
authors.lastname: Abedi
authors.forename: Gerhard
authors.initials: G
authors.lastname: Bauer
authors.forename: Jan A
authors.initials: JA
authors.lastname: Nolta
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Anderson
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000276:immunology; D064987:Cell- and Tissue-Based Therapy; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D015658:HIV Infections / Q000235:genetics / Q000276:immunology* / Q000628:therapy*; D015497:HIV-1 / Q000276:immunology* / Q000502:physiology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000276:immunology* / Q000821:virology; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D051379:Mice; D014161:Transduction, Genetic
chemical_list: D018952:Antigens, CD34