Highly active antiretroviral therapies (HAART) is incapable of fully eradicating virus within an adult patient, due to the persistence of latently infected resting CD4+ T lymphocytes and infected macrophages and monocytes, which may produce low levels of viremia without dying. Recently, there has been a growing interest in using gene therapy as an alternative to HAART, particularly with the recent breakthrough of the Berlin patient, who was "functionally cured" of HIV after receiving an allogeneic bone marrow transplant with CCR5-defective hematopoietic stem cells. Indeed, we have conducted the only clinical trial to date to test an anti-HIV RNAi therapeutic, which required the ex vivo delivery of a lentiviral vector in autologous CD34+ hematopoietic stem cells for patients with AIDS-related lymphoma. However, for RNAi-based therapeutics to become available for the vast majority of HIV patients, alternative systemic delivery methods must be developed. In this project, we will test three promising methods for delivering anti-HIV siRNAs: an anti-gp120 RNA aptamer-siRNA conjugate for targeted delivery into cells actively infected with HIV;an anti-CD4 RNA aptamer-siRNA conjugate for targeted delivery into CD4+ T cells and macrophages;and a cationic PAMAM dendrimer-siRNA nanoparticle for systemic delivery.
For Aim 1, we will elucidate the molecular interactions between the anti-gp120 aptamer and HIV gp120 protein using surface plasmon resonance, X-ray crystallography, and electron microscopy.
In Aim2 of this project, we will use positron emission tomography and near-infrared fluorescence imaging to measure the dose- dependent biodistribution and whole-body clearance of our RNA-based therapeutics in a humanized mouse model.
In Aim 3, we will conduct a series of in vivo experiments in HIV-infected humanized mice to test the efficacy of individual and combination RNA-based therapies. These experiments will test siRNAs that target endogenous genes required for HIV infection (prophylactic and entry inhibitor siRNAs), mechanisms required for the persistence of latency (anti-latency siRNAs), and genes required for cell survival (cytotoxic siRNAs). Hence, we will test a comprehensive, RNA-based "shock and kill" strategy that aims to purge the latent reservoir, eliminate the cells actively infected with HIV, and protect cells from new infection. In addition to standard assays for detecting HIV viremia, we will use droplet digital PCR to measure the levels of integrated provirus in treated and non-treated animals. In summary, this project represents an innovative approach for suppressing and potentially eradicating HIV infection. Moreover, the aims of this pre-clinical study will establish an important benchmark for evaluating RNAi- based therapy as a treatment for HIV.

Public Health Relevance

Recent advances in RNAi-based drugs have broadened the scope of therapeutic targets for a variety of human disease. We have designed a new systemic delivery approach for a cocktail of anti-HIV siRNAs using chemically synthesized, ligand-specific RNA aptamers or cationic PAMAM dendrimer nanoparticles. We will test these RNA therapies in a pre-clinical investigation in humanized mice by monitoring the biodistribution, whole-body clearance, and off-target toxicity. In addition, we will perform functional analyses in HIV-infecte mice to examine the potential eradication of latent reservoirs and prophylactic treatment of siRNAs pre- and post-HIV challenge.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
Project #
Application #
Study Section
AIDS Discovery and Development of Therapeutics Study Section (ADDT)
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
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
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; 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
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
Sakurai, Kumi; Amarzguioui, Mohammed; Kim, Dong-Ho et al. (2011) A role for human Dicer in pre-RISC loading of siRNAs. Nucleic Acids Res 39:1510-25
Zhou, Jiehua; Li, Haitang; Zhang, Jane et al. (2011) Development of cell-type specific anti-HIV gp120 aptamers for siRNA delivery. J Vis Exp :

Showing the most recent 10 out of 112 publications