HIV/AIDS continues to be a major threat to human health. The use of combinations of small molecule drugs in highly active anti-retroviral therapy (HAART) to stop or thwart HIV propagation ahs had a major impact on delaying the progression from HIV-1 infection to the development of AIDS. Despite this progress, there are problems associated with a lifelong use of anti-viral drug therapy. These include toxicity, the emergence of virus resistant to multiple drugs, and the cost of a daily medication. The proposed studies take advantage of recent advances in gene therapy for the treatment of HIV-1 infection. Gene therapy of human blood progenitor cells can be considered as a potential replacement or supplement to the current anti-HIV-1 therapies. An outcome of the previous funding period was the development of a three pronged genetic approach for inhibiting viral entry and replication. This triple small RNA gene therapy has been tested for feasibility and safety in a first in human blood stem cell gene therapy trial in AIDS/lymphoma patients. Based upon the results of this four patient trial we believe it is imperative to progress to new and different small RNA anti-viral combinations. The proposed studies take advantage of a set of novel inhibitors of HIV-1 infection which use different mechanisms of action. We hypothesize that such combinations will prevent the emergence of viral resistant mutants and at the same time be non-toxic.
Specific Aim 1 will select for in vitro evolved "RNA aptamers that selectively bind HIV-1 integrase and block integrase function". The aptamers will be expressed in hematopoietic cells as part of a combinatorial approach designed to block viral infection.
Specific Aim 2 will "combine nucleolar localizing anti-HIV RNAs, microRNA mimics, tRNA-anti CCR5 shRNA, U5 tRNA shRNA fusions and the anti-integrase aptamers in a lentiviral vector for transduction of hematopoietic progenitor cells". The results from this research will provide novel combinations of anti-HIV RNAs for functional testing in vivo.
Specific Aim 3 addresses the "in vivo analyses of combinatorial constructs in the humanized Rag2-/- 3c-/- (RAG-hu) mouse model". This humanized mouse model represents a novel and facile system for testing the efficacy and safety of anti-HIV gene therapy strategies since gene modified human hematopoietic stem and progenitor cells can engraft and differentiate into hematopoietic lineages that are infectible by HIV-1 in vivo. Overall, this program is designed to be a path towards bringing new small RNA based gene therapy combinations to the clinic for use in future human gene therapy applications.

Public Health Relevance

Combinatorial anti-HIV small drug therapies have significantly prolonged the time to progression to AIDS for HIV-1 infected individuals. Nevertheless there are toxicity, viral resistance and compliance problems associated with a lifetime of daily use of these drug combinations. The proposed studies capitalize upon the safety and feasibility of a study exploring combinatorial small RNA gene therapy for the treatment of HIV-1 infection. Combinations of novel anti-HIV-1 small RNAs will be tested for their ability to provide long- term inhibition of HIV-1 infection in cultured human blood cells and in a humanized mouse model. The long term goal of these studies is the establishment of potent, non-toxic combinations of anti-HIV small RNAs for use in gene therapy treatment of HIV-1 infection.

National Institute of Health (NIH)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Voulgaropoulou, Frosso
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
City of Hope/Beckman Research Institute
United States
Zip Code
Chung, Janet; Scherer, Lisa J; Gu, Angel et al. (2014) Optimized lentiviral vectors for HIV gene therapy: multiplexed expression of small RNAs and inclusion of MGMT(P140K) drug resistance gene. Mol Ther 22:952-63
Zhou, Jiehua; Rossi, John (2014) Cell-type-specific aptamer and aptamer-small interfering RNA conjugates for targeted human immunodeficiency virus type 1 therapy. J Investig Med 62:914-9
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
Zhou, Jiehua; Shum, Ka-To; Burnett, John C et al. (2013) Nanoparticle-Based Delivery of RNAi Therapeutics: Progress and Challenges. Pharmaceuticals (Basel) 6:85-107
Ehsani, Ali; Alluin, Jessica V; Rossi, John J (2013) Cell cycle abnormalities associated with differential perturbations of the human U5 snRNP associated U5-200kD RNA helicase. PLoS One 8:e62125
Chung, Janet; DiGiusto, David L; Rossi, John J (2013) Combinatorial RNA-based gene therapy for the treatment of HIV/AIDS. Expert Opin Biol Ther 13:437-45
Cillo, Anthony R; Krishnan, Amrita; Mitsuyasu, Ronald T et al. (2013) Plasma viremia and cellular HIV-1 DNA persist despite autologous hematopoietic stem cell transplantation for HIV-related lymphoma. J Acquir Immune Defic Syndr 63:438-41
Burnett, John C; Rossi, John J (2012) RNA-based therapeutics: current progress and future prospects. Chem Biol 19:60-71
Sun, Guihua; Li, Haitang; Wu, Xiwei et al. (2012) Interplay between HIV-1 infection and host microRNAs. Nucleic Acids Res 40:2181-96
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

Showing the most recent 10 out of 65 publications