SiRNA Gene Therapy for HIV/AIDS

Summary

Principal Investigator: RAMESH K AKKINA
Abstract: [unreadable] DESCRIPTION (provided by applicant): 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. [unreadable] [unreadable]
Funding Period: 2004-04-01 - 2010-03-31
more information: NIH RePORT

Top Publications

  1. pmc Derivation of normal macrophages from human embryonic stem (hES) cells for applications in HIV gene therapy
    Joseph S Anderson
    Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado 80523, USA
    Retrovirology 3:24. 2006
  2. pmc Systemic administration of combinatorial dsiRNAs via nanoparticles efficiently suppresses HIV-1 infection in humanized mice
    Jiehua Zhou
    Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, City of Hope, Duarte, California 91010, USA
    Mol Ther 19:2228-38. 2011
  3. pmc Humanized Rag1-/- γc-/- mice support multilineage hematopoiesis and are susceptible to HIV-1 infection via systemic and vaginal routes
    Ramesh Akkina
    Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, United States of America
    PLoS ONE 6:e20169. 2011
  4. pmc A topical microbicide gel formulation of CCR5 antagonist maraviroc prevents HIV-1 vaginal transmission in humanized RAG-hu mice
    C Preston Neff
    Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America
    PLoS ONE 6:e20209. 2011
  5. pmc Thymic plasmacytoid dendritic cells are susceptible to productive HIV-1 infection and efficiently transfer R5 HIV-1 to thymocytes in vitro
    Vanessa A Evans
    Monash University, Department of Medicine, Central and Eastern Clinical School, Alfred Campus, Commercial Rd, Melbourne, Victoria 3004, Australia
    Retrovirology 8:43. 2011
  6. pmc An aptamer-siRNA chimera suppresses HIV-1 viral loads and protects from helper CD4(+) T cell decline in humanized mice
    Charles Preston Neff
    Department of Microbiology, Immunology and Pathology, Colorado State University, 1619 Campus Delivery, Fort Collins, CO 80523, USA
    Sci Transl Med 3:66ra6. 2011
  7. pmc Oral pre-exposure prophylaxis by anti-retrovirals raltegravir and maraviroc protects against HIV-1 vaginal transmission in a humanized mouse model
    C Preston Neff
    Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America
    PLoS ONE 5:e15257. 2010
  8. pmc Humanized Rag2(-/-)gammac(-/-) (RAG-hu) mice can sustain long-term chronic HIV-1 infection lasting more than a year
    Bradford K Berges
    Department of Microbiology, Immunology, and Pathology, Colorado State University, 329 Pathology, Fort Collins, CO 80523, USA
    Virology 397:100-3. 2010
  9. ncbi 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
    Joseph Anderson
    Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA
    Hum Gene Ther 19:217-28. 2008
  10. pmc Mucosal transmission of R5 and X4 tropic HIV-1 via vaginal and rectal routes in humanized Rag2-/- gammac -/- (RAG-hu) mice
    Bradford K Berges
    Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA
    Virology 373:342-51. 2008

Detail Information

Publications14

  1. pmc Derivation of normal macrophages from human embryonic stem (hES) cells for applications in HIV gene therapy
    Joseph S Anderson
    Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado 80523, USA
    Retrovirology 3:24. 2006
    ....
  2. pmc Systemic administration of combinatorial dsiRNAs via nanoparticles efficiently suppresses HIV-1 infection in humanized mice
    Jiehua Zhou
    Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, City of Hope, Duarte, California 91010, USA
    Mol Ther 19:2228-38. 2011
    ..The dendrimer delivery approach therefore represents a promising method for systemic delivery of combinations of siRNAs for treatment of HIV-1 infection...
  3. pmc Humanized Rag1-/- γc-/- mice support multilineage hematopoiesis and are susceptible to HIV-1 infection via systemic and vaginal routes
    Ramesh Akkina
    Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, United States of America
    PLoS ONE 6:e20169. 2011
    ..Thus these mice can be further exploited for studying human pathogens that infect the human hematopoietic system in an in vivo setting...
  4. pmc A topical microbicide gel formulation of CCR5 antagonist maraviroc prevents HIV-1 vaginal transmission in humanized RAG-hu mice
    C Preston Neff
    Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America
    PLoS ONE 6:e20209. 2011
    ....
  5. pmc Thymic plasmacytoid dendritic cells are susceptible to productive HIV-1 infection and efficiently transfer R5 HIV-1 to thymocytes in vitro
    Vanessa A Evans
    Monash University, Department of Medicine, Central and Eastern Clinical School, Alfred Campus, Commercial Rd, Melbourne, Victoria 3004, Australia
    Retrovirology 8:43. 2011
    ..Transfer of productive HIV-1 infection from thymic mDC and pDC was determined by culturing these DC subsets either alone or with sorted thymocytes...
  6. pmc An aptamer-siRNA chimera suppresses HIV-1 viral loads and protects from helper CD4(+) T cell decline in humanized mice
    Charles Preston Neff
    Department of Microbiology, Immunology and Pathology, Colorado State University, 1619 Campus Delivery, Fort Collins, CO 80523, USA
    Sci Transl Med 3:66ra6. 2011
    ..The aptamer thus acts as a broad-spectrum HIV-neutralizing agent and an siRNA delivery vehicle. The combined aptamer-siRNA agent provides an attractive, nontoxic therapeutic approach for treatment of HIV infection...
  7. pmc Oral pre-exposure prophylaxis by anti-retrovirals raltegravir and maraviroc protects against HIV-1 vaginal transmission in a humanized mouse model
    C Preston Neff
    Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America
    PLoS ONE 5:e15257. 2010
    ..Based on these results both these drugs show great promise for further development as orally administered PrEPs...
  8. pmc Humanized Rag2(-/-)gammac(-/-) (RAG-hu) mice can sustain long-term chronic HIV-1 infection lasting more than a year
    Bradford K Berges
    Department of Microbiology, Immunology, and Pathology, Colorado State University, 329 Pathology, Fort Collins, CO 80523, USA
    Virology 397:100-3. 2010
    ..These studies provide a chronic HIV-1 infection humanized mouse model that can be used to dissect viral latency, long-term drug evaluation and immune-based therapies...
  9. ncbi 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
    Joseph Anderson
    Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA
    Hum Gene Ther 19:217-28. 2008
    ..These results demonstrate the efficacy of TRIM5alpha-HRH in a stem cell setting and its further advancement for use in gene therapy applications...
  10. pmc Mucosal transmission of R5 and X4 tropic HIV-1 via vaginal and rectal routes in humanized Rag2-/- gammac -/- (RAG-hu) mice
    Bradford K Berges
    Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA
    Virology 373:342-51. 2008
    ....
  11. ncbi Complete knockdown of CCR5 by lentiviral vector-expressed siRNAs and protection of transgenic macrophages against HIV-1 infection
    J Anderson
    Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523, USA
    Gene Ther 14:1287-97. 2007
    ..Thus the above anti-CCR5 siRNAs are among the most effective demonstrated to date and are very promising candidates for clinical applications...
  12. ncbi Safety and efficacy of a lentiviral vector containing three anti-HIV genes--CCR5 ribozyme, tat-rev siRNA, and TAR decoy--in SCID-hu mouse-derived T cells
    Joseph Anderson
    Department of Microbiology, Immunology and Pathology, Colorado State University, Fort, Collins, Colorado 80523, USA
    Mol Ther 15:1182-8. 2007
    ..These important attributes of this combinatorial vector show its promise as an excellent candidate for use in human clinical trials...
  13. pmc HIV-1 infection and CD4 T cell depletion in the humanized Rag2-/-gamma c-/- (RAG-hu) mouse model
    Bradford K Berges
    Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA
    Retrovirology 3:76. 2006
    ..Thus, this model shows considerable promise to study long-term in vivo HIV infection and pathogenesis...
  14. pmc Functional in vivo delivery of multiplexed anti-HIV-1 siRNAs via a chemically synthesized aptamer with a sticky bridge
    Jiehua Zhou
    Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, Duarte, California 91010, USA
    Mol Ther 21:192-200. 2013
    ..Collectively, these data demonstrate a facile, targeted approach for combinatorial delivery of antiviral and host DsiRNAs for HIV-1 therapy in vivo...