In the current proposal we are characterizing how DNA recognition by cytosolic DNA sensors coordinates activation of both the inflammasome and type I Interferon (IRF3) activated antiviral cascades. We are proposing the cytosolic DNA sensor protein AIM2 is pivotal to activation of both cascades. Using recombinant adenovirus vector infection of macrophage cell lines or primary cells, we are determining how triggering the DNA sensing machinery signals activation of the IRF3 type I interferon response and influences maturation of these antigen presenting cells. Based on preliminary data, we show that one of the essential downstream targets of the DNA sensor response is the adaptor protein STING. The scaffolds that support DNA sensor co-activation of inflammasome/STING targets will be examined. The current proposal presents the hypothesis that AIM2 is functioning to balance the antigen-presenting cell antiviral response. We have found that AIM2 which is an interferon inducible gene is also involved in regulating type I interferon sensitivity through STAT1. We are proposing an AIM2/STAT1 regulatory loop functions to control the inflammatory/IFN sensitivity of target immune sentinel cells. The studies presented in this proposal are targeting the immune recognition response pathways that are at the heart of the innate and adaptive immune response to recombinant adenovirus vectors. We are proposing these pathways are operating in a specific manner in immune cells. The knowledge gained from these studies will contribute to enhancing vaccine, gene therapy, and anticancer therapeutic applications and they will contribute to our ability to develop new treatments for viral infections and host DNA dependent autoimmune diseases.

Public Health Relevance

Immune stimulating (is) DNA directly impacts immune function in DNA vaccines, DNA dependent autoimmune diseases and antiviral Immune responses. isDNA is a new and poorly understood area of the host (antiviral) immune response. In the current proposal we are investigating the DNA sensing networks triggered by rAdV infection of murine macrophages. Our research in this exciting and important area will contribute to immune modification strategies able to enhance the potency of DNA vaccines, enhance DNA vector gene transfer, or suppress DNA induced inflammatory diseases.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01AI094050-04
Application #
8686730
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Palker, Thomas J
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Weill Medical College of Cornell University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
City
New York
State
NY
Country
United States
Zip Code
10065
Lam, Eric; Stein, Saskia; Falck-Pedersen, Erik (2014) Adenovirus detection by the cGAS/STING/TBK1 DNA sensing cascade. J Virol 88:974-81
Janovitz, Tyler; Oliveira, Thiago; Sadelain, Michel et al. (2014) Highly divergent integration profile of adeno-associated virus serotype 5 revealed by high-throughput sequencing. J Virol 88:2481-8
Janovitz, Tyler; Sadelain, Michel; Falck-Pedersen, Erik (2014) Adeno-associated virus type 2 preferentially integrates single genome copies with defined breakpoints. Virol J 11:15
Janovitz, Tyler; Klein, Isaac A; Oliveira, Thiago et al. (2013) High-throughput sequencing reveals principles of adeno-associated virus serotype 2 integration. J Virol 87:8559-68
Stein, Saskia C; Falck-Pedersen, Erik (2012) Sensing adenovirus infection: activation of interferon regulatory factor 3 in RAW 264.7 cells. J Virol 86:4527-37