The recently described RNA interference (RNAi) phenomenon mediated by small interfering RNAs (siRNAs) is highly sequence specific gene silencing mechanism. Recent studies established the remarkable potency of siRNAs in suppressing HIV-1 replication in vitro and highlighted its potential as 3owerful tool for gene therapy. Translation of this promising new technology into the clinic requires stable introduction of the siRNA genes into hematopoietic stem cells and derivation of viral resistant T cells and macrophages. Using in vitro assays and a unique SCID-hu mouse in vivo model that harbors transplanted human tissue, we recently showed that CD34 hematopoietic progenitor cells transduced with anti-HIV siRNAs, ribozymes and RNA decoys could be differentiated into virus resistant T cells and macrophages. These proofs of concept studies paved the way to evaluate novel approaches in an in vivo setting. Several new developments have also occurred recently in the areas of stem cell biology, lentiviral vectors, and in vivo modeling with direct relevance to HIV gene therapy. In the current proposal our goal is to build upon our recent progress. Our specific objectives are: 1. Evaluate the synergistic efficacy of novel combinatorial siRNA constructs targeted to different stages of HIV-1 life cycle in vitro and analyze their mechanism of action. 2. Transduce siRNAs via lentiviral vectors into CD34 hematopoietic progenitor cells to derive HIV-1 resistant macrophages and investigate the mechanism of their action in differentiated cells. 3. Determine the in vivo protective effects of different anti-HIV-1 siRNAs, either individually or in combination, in SCID-hu mice thy/liv grafts against HIV-1 challenge and also assess the functional competence of transgenic T cells. 4. Transduce siRNA constructs into the newly described CD34+/KDR+ primitive hematopoietic progenitor cells and derive macrophages in vitro and thymocytes in vivo, and evaluate their HIV-1 resistance. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI057066-03
Application #
7034643
Study Section
Special Emphasis Panel (ZRG1-AARR-E (03))
Program Officer
Black, Paul L
Project Start
2004-04-01
Project End
2009-03-31
Budget Start
2006-04-01
Budget End
2007-03-31
Support Year
3
Fiscal Year
2006
Total Cost
$318,584
Indirect Cost
Name
Colorado State University-Fort Collins
Department
Microbiology/Immun/Virology
Type
Schools of Veterinary Medicine
DUNS #
785979618
City
Fort Collins
State
CO
Country
United States
Zip Code
80523
Zhou, Jiehua; Neff, C Preston; Swiderski, Piotr et al. (2013) Functional in vivo delivery of multiplexed anti-HIV-1 siRNAs via a chemically synthesized aptamer with a sticky bridge. Mol Ther 21:192-200
Neff, Charles Preston; Zhou, Jiehua; Remling, Leila et al. (2011) An aptamer-siRNA chimera suppresses HIV-1 viral loads and protects from helper CD4(+) T cell decline in humanized mice. Sci Transl Med 3:66ra6
Evans, Vanessa A; Lal, Luxshimi; Akkina, Ramesh et al. (2011) Thymic plasmacytoid dendritic cells are susceptible to productive HIV-1 infection and efficiently transfer R5 HIV-1 to thymocytes in vitro. Retrovirology 8:43
Neff, C Preston; Kurisu, Theresa; Ndolo, Thomas et al. (2011) A topical microbicide gel formulation of CCR5 antagonist maraviroc prevents HIV-1 vaginal transmission in humanized RAG-hu mice. PLoS One 6:e20209
Zhou, Jiehua; Neff, C Preston; Liu, Xiaoxuan et al. (2011) Systemic administration of combinatorial dsiRNAs via nanoparticles efficiently suppresses HIV-1 infection in humanized mice. Mol Ther 19:2228-38
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Berges, Bradford K; Akkina, Sarah R; Remling, Leila et al. (2010) Humanized Rag2(-/-)gammac(-/-) (RAG-hu) mice can sustain long-term chronic HIV-1 infection lasting more than a year. Virology 397:100-3
Neff, C Preston; Ndolo, Thomas; Tandon, Apurva et al. (2010) Oral pre-exposure prophylaxis by anti-retrovirals raltegravir and maraviroc protects against HIV-1 vaginal transmission in a humanized mouse model. PLoS One 5:e15257
Anderson, Joseph; Akkina, Ramesh (2008) Human immunodeficiency virus type 1 restriction by human-rhesus chimeric tripartite motif 5alpha (TRIM 5alpha) in CD34(+) cell-derived macrophages in vitro and in T cells in vivo in severe combined immunodeficient (SCID-hu) mice transplanted with human fe Hum Gene Ther 19:217-28
Berges, Bradford K; Akkina, Sarah R; Folkvord, Joy M et al. (2008) Mucosal transmission of R5 and X4 tropic HIV-1 via vaginal and rectal routes in humanized Rag2-/- gammac -/- (RAG-hu) mice. Virology 373:342-51

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