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)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-AARR-E (02))
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
Pang, Ka Ming; Castanotto, Daniela; Li, Haitang et al. (2018) Incorporation of aptamers in the terminal loop of shRNAs yields an effective and novel combinatorial targeting strategy. Nucleic Acids Res 46:e6
Reebye, Vikash; Huang, Kai-Wen; Lin, Vivian et al. (2018) Gene activation of CEBPA using saRNA: preclinical studies of the first in human saRNA drug candidate for liver cancer. Oncogene 37:3216-3228
Zhou, Jiehua; Lazar, Daniel; Li, Haitang et al. (2018) Receptor-targeted aptamer-siRNA conjugate-directed transcriptional regulation of HIV-1. Theranostics 8:1575-1590
Yoon, Sorah; Wu, Xiwei; Armstrong, Brian et al. (2018) An RNA Aptamer Targeting the Receptor Tyrosine Kinase PDGFR? Induces Anti-tumor Effects through STAT3 and p53 in Glioblastoma. Mol Ther Nucleic Acids 14:131-141
Satheesan, Sangeetha; Li, Haitang; Burnett, John C et al. (2018) HIV Replication and Latency in a Humanized NSG Mouse Model during Suppressive Oral Combinational Antiretroviral Therapy. J Virol 92:
Hori, Shin-Ichiro; Herrera, Alberto; Rossi, John J et al. (2018) Current Advances in Aptamers for Cancer Diagnosis and Therapy. Cancers (Basel) 10:
Song, Min-Sun; Rossi, John J (2017) Molecular mechanisms of Dicer: endonuclease and enzymatic activity. Biochem J 474:1603-1618
Yoon, Sorah; Armstrong, Brian; Habib, Nagy et al. (2017) Blind SELEX Approach Identifies RNA Aptamers That Regulate EMT and Inhibit Metastasis. Mol Cancer Res 15:811-820
Zhou, Jiehua; Rossi, John (2017) Aptamers as targeted therapeutics: current potential and challenges. Nat Rev Drug Discov 16:181-202
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

Showing the most recent 10 out of 106 publications