Human immunodeficiency virus type 1 (HIV-1) emerged as a major human disease approximately 30 years ago, with roughly 70% of the current cases in sub-Saharan Africa and little hope for an efficacious vaccine in the near future. Alternative strategies to inhibit HIV-1 replication or purge viral resen/oirs are tremendously desirable. Several new and innovative strategies are proposed here and center on delivering small non-coding RNAs and an RNA targeted humanized protein complex capable of targeting the excision of integrated forms of HIV-1 or CCRS to specific cells. We hypothesize that it is possible to specifically target HIV-1 infected or relevant cell types in vivo and stably silence or excise HIV-1 or CCRS from these cells. The result of this targeted suppression/excision is a loss of HIV-1 fidelity and a functional cure. We will test our hypothesis using the recently developed and well-validated Rag2-/- yc-/- (RAG-hu) mouse model. We propose 3 aims which will test our hypothesis;
aim 1 contrast the various conditionally replicating vector systems for suppression and/or excision of HIV-1 and CCRS in the RAG-hu mouse model, aim 2 determine the ability of aptamer targeted minigenes or siRNAs to suppress and/or target the excision of HIV-1 or CCRS in vivo, and in aim 3 we will determine the ability of nanoparticle targeted minigenes and the Pddlp excision complex to suppress and target the excision of HlV-1 or CCRS in vivo. We envision one of these three different delivery platforms to prove efficacious at targeting HlV-1 or CCRS in target cells and significantly affecting viral replication and evolution, leading ultimately to functional cure (a state where viral replication and mutation is inhibited). Notably, data generated from the in vivo core can be translated efficiently into human based trials depending on the resultant findings.

Public Health Relevance

This project will determine ability of three different cell targeted delivery strategies to selectively inhibit HIV-1 or CCRS expression in vivo and specifically affect the pathogenicity and infectivity of HIV-1.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI099783-02
Application #
8451993
Study Section
Special Emphasis Panel (ZAI1-RB-A)
Project Start
Project End
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
2
Fiscal Year
2013
Total Cost
$302,501
Indirect Cost
$85,437
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Trakman, Laura; Hewson, Chris; Burdach, Jon et al. (2016) RNA Directed Modulation of Phenotypic Plasticity in Human Cells. PLoS One 11:e0152424
Lazar, Daniel C; Morris, Kevin V; Saayman, Sheena M (2016) The emerging role of long non-coding RNAs in HIV infection. Virus Res 212:114-26
Fortes, Puri; Morris, Kevin V (2016) Long noncoding RNAs in viral infections. Virus Res 212:1-11
Saayman, Sheena M; Lazar, Daniel C; Scott, Tristan A et al. (2016) Potent and Targeted Activation of Latent HIV-1 Using the CRISPR/dCas9 Activator Complex. Mol Ther 24:488-98
Hewson, Chris; Morris, Kevin V (2016) Form and Function of Exosome-Associated Long Non-coding RNAs in Cancer. Curr Top Microbiol Immunol 394:41-56
Saayman, Sheena; Ali, Stuart A; Morris, Kevin V et al. (2015) The therapeutic application of CRISPR/Cas9 technologies for HIV. Expert Opin Biol Ther 15:819-30
Roberts, Thomas C; Hart, Jonathan R; Kaikkonen, Minna U et al. (2015) Quantification of nascent transcription by bromouridine immunocapture nuclear run-on RT-qPCR. Nat Protoc 10:1198-211
Yu, Albert D; Wang, Zichen; Morris, Kevin V (2015) Long noncoding RNAs: a potent source of regulation in immunity and disease. Immunol Cell Biol 93:277-83
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
Vadie, Nadia; Saayman, Sheena; Lenox, Alexandra et al. (2015) MYCNOS functions as an antisense RNA regulating MYCN. RNA Biol 12:893-9

Showing the most recent 10 out of 33 publications