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.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI042552-17
Application #
8683060
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Voulgaropoulou, Frosso
Project Start
1998-01-01
Project End
2016-06-30
Budget Start
2014-07-01
Budget End
2015-06-30
Support Year
17
Fiscal Year
2014
Total Cost
Indirect Cost
Name
City of Hope/Beckman Research Institute
Department
Type
DUNS #
City
Duarte
State
CA
Country
United States
Zip Code
91010
Takahashi, Mayumi; Wu, Xiwei; Ho, Michelle et al. (2016) High throughput sequencing analysis of RNA libraries reveals the influences of initial library and PCR methods on SELEX efficiency. Sci Rep 6:33697
Bobbin, Maggie L; Rossi, John J (2016) RNA Interference (RNAi)-Based Therapeutics: Delivering on the Promise? Annu Rev Pharmacol Toxicol 56:103-22
Zhou, Jiehua; Rossi, John J (2016) Evolution of Cell-Type-Specific RNA Aptamers Via Live Cell-Based SELEX. Methods Mol Biol 1421:191-214
Daniels, Sylvanne M; Sinck, Lucile; Ward, Natalie J et al. (2015) HIV-1 RRE RNA acts as an RNA silencing suppressor by competing with TRBP-bound siRNAs. RNA Biol 12:123-35
Zhou, Jiehua; Satheesan, Sangeetha; Li, Haitang et al. (2015) Cell-specific RNA aptamer against human CCR5 specifically targets HIV-1 susceptible cells and inhibits HIV-1 infectivity. Chem Biol 22:379-90
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
Chadalavada, Deepti; Adamson, Trinka W; Burnett, John C et al. (2014) Irradiated compared with nonirradiated NSG mice for the development of a human B-cell lymphoma model. Comp Med 64:179-85
Zhang, Xizhe; Li, Haitang; Burnett, John C et al. (2014) The role of antisense long noncoding RNA in small RNA-triggered gene activation. RNA 20:1916-28
Zhou, Jiehua; Rossi, John J (2014) Cell-type-specific, Aptamer-functionalized Agents for Targeted Disease Therapy. Mol Ther Nucleic Acids 3:e169
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

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