The specificity and potency of targeted gene knockdown through RNA interference (RNAi) has generated tremendous excitement for utilizing this ubiquitous pathway in the clinic. Our studies investigate RNAi as a potential microbicide to target sexually transmitted diseases including HSV-2 and HIV-1. HSV-2 infects ~ 20% of adults in the US and up to 90% in parts of sub-Saharan Africa. It is associated with high morbidity and there is no cure or prophylactic treatment. HSV-2 is also a major co-factor for HIV-1. Therefore, a microbicide to HSV-2 could also have a significant impact on slowing HIV-1 transmission. Using a mouse model of vaginal HSV-2 infection, we were the first group to show that vaginal application of siRNAs targeting HSV-2 viral genes or host-encoded viral entry receptor genes could protect mice from HSV-2 infection. These findings were encouraging, but we encountered issues that would need to be addressed for siRNAs to be developed for clinical use. Problems included toxicity of a cationic lipid used to complex the siRNAs, transient protection conferred by virus-specific siRNAs, and siRNA instability in vaginal washes. Each of these problems was addressed in a follow-up study. In this proposal we will undertake a comprehensive analysis of HSV-2 gene targeted siRNAs to maximize their specific silencing ability, and minimize any off-target effects. Using the murine HSV-2 infection model we will determine whether: (i) immune responses, induced either by the siRNA or through viral challenge affect siRNA-mediated protection;(ii) enhanced incorporation of siRNA into RISC can result in more potent or durable protection;and (iii) efficient siRNA uptake and intracellular localization in the vaginal mucosa requires the expression of a specific receptor.

Public Health Relevance

Sexually transmitted diseases, such as HSV-2 and HIV-1 represent a major public health burden. We have been using RNA interference (RNAi) to knockdown the expression of HSV-2 viral genes and the host receptors they use to access cellular targets. RNAi-treated mice are protected from HSV-2 transmission. The goal of this proposal is to optimize RNAi reagents for clinical use against HSV-2 infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI099567-01
Application #
8276523
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Deal, Carolyn D
Project Start
2012-03-19
Project End
2017-02-28
Budget Start
2012-03-19
Budget End
2013-02-28
Support Year
1
Fiscal Year
2012
Total Cost
$416,667
Indirect Cost
$166,667
Name
Albert Einstein College of Medicine
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
110521739
City
Bronx
State
NY
Country
United States
Zip Code
10461
Katakowski, Joseph A; Mukherjee, Gayatri; Wilner, Samantha E et al. (2016) Delivery of siRNAs to Dendritic Cells Using DEC205-Targeted Lipid Nanoparticles to Inhibit Immune Responses. Mol Ther 24:146-55
Lauvau, Grégoire; Boutet, Marie; Williams, Tere M et al. (2016) Memory CD8(+) T Cells: Innate-Like Sensors and Orchestrators of Protection. Trends Immunol 37:375-85
Brojatsch, Jürgen; Lima Jr, Heriberto; Palliser, Deborah et al. (2015) Distinct cathepsins control necrotic cell death mediated by pyroptosis inducers and lysosome-destabilizing agents. Cell Cycle 14:964-72
Tosti, Elena; Katakowski, Joseph A; Schaetzlein, Sonja et al. (2014) Evolutionarily conserved genetic interactions with budding and fission yeast MutS identify orthologous relationships in mismatch repair-deficient cancer cells. Genome Med 6:68
Soudja, Saïdi M'Homa; Chandrabos, Ceena; Yakob, Ernest et al. (2014) Memory-T-cell-derived interferon-γ instructs potent innate cell activation for protective immunity. Immunity 40:974-88
Wengerter, Brian C; Katakowski, Joseph A; Rosenberg, Jacob M et al. (2014) Aptamer-targeted antigen delivery. Mol Ther 22:1375-87
Magalhães, Maria L B; Byrom, Michelle; Yan, Amy et al. (2012) A general RNA motif for cellular transfection. Mol Ther 20:616-24