Host exposure to microbial pathogens such as viruses, bacteria and fungi trigger the activation of innate immune responses, including the production of type I interferon (IFN) that galvanize early host defense mechanisms as well as invigorate adaptive immune responses involving cytotoxic T cell activity and antibody production. The recognition of pathogenic microbes and the triggering of the innate immune cascade has become the subject of intense research over the past few years. Particular attention has recently focused on the role of the Toll-like receptors (TLRs), which have emerged as key molecules largely expressed by dendritic cells and macrophages, that are responsible for recognizing conserved components of pathogenic microorganisms (referred to as pathogen associated molecular patterns -PAMPs)- and which trigger the production of IFN. However, it has recently been discovered that critically important TLR-independent mechanisms also exist to thwart pathogen infection. Instrumental in these responses are a family of DExD/H helicases that appear critical for effective defense against virus infection. However, the mechanisms of TLR-independent signaling remain complex, with downstream molecular components responsible for facilitating the production of IFN remaining to be identified. Here, we have isolated a new molecule that appears to play a significant role in the regulation of innate immune signaling and which is critical for the production of IFN in response to pathogen infection. For this proposal we thus aim to study the importance of this molecule in innate immune helicase action. Essentially, we aim to: I: determine the importance of DExD/H helicases in innate immunity to pathogen infection, including mechanisms of action. II: To potentially determine the mechanisms of innate signaling mediated by the DExD/H helicases involved in pathogen recognition, we have effectively screened for novel molecules which activate the production of type I interferon (IFN), required for effective innate immunity.
We aim to further characterize the importance of such molecules in innate signaling and aim to determine their mechanisms of action. These objectives will shed significant insight into the mechanisms of innate immune signaling and generate concepts useful for the generation of new vaccine and immunotherapeutic strategies.

Public Health Relevance

Our grant involves attempting to understand how the body recognizes diseases caused by viruses and bacteria. The proposal also aims to understand how the body induces an appropriate immune response following infection. By understanding these processes, we will gain insight into causes of disease and anticipate being able to develop new vaccine and therapeutic strategies. The study of LGP2 and STING and their interaction with other components in the pathway will provide significant insight about the molecular mechanism of RIG-I/MDA5-mediated IFN production and anti-viral innate immunity and thus is of high significance.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI079336-05
Application #
8306174
Study Section
Innate Immunity and Inflammation Study Section (III)
Program Officer
Palker, Thomas J
Project Start
2008-08-01
Project End
2014-02-28
Budget Start
2012-08-01
Budget End
2014-02-28
Support Year
5
Fiscal Year
2012
Total Cost
$374,888
Indirect Cost
$129,863
Name
University of Miami School of Medicine
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
052780918
City
Coral Gables
State
FL
Country
United States
Zip Code
33146
Abe, Takayuki; Barber, Glen N (2014) Cytosolic-DNA-mediated, STING-dependent proinflammatory gene induction necessitates canonical NF-?B activation through TBK1. J Virol 88:5328-41
Konno, Hiroyasu; Konno, Keiko; Barber, Glen N (2013) Cyclic dinucleotides trigger ULK1 (ATG1) phosphorylation of STING to prevent sustained innate immune signaling. Cell 155:688-98
Abe, Takayuki; Harashima, Ai; Xia, Tianli et al. (2013) STING recognition of cytoplasmic DNA instigates cellular defense. Mol Cell 50:5-15
Barber, Glen N (2011) Innate immune DNA sensing pathways: STING, AIMII and the regulation of interferon production and inflammatory responses. Curr Opin Immunol 23:10-20
Martinez, Jennifer; Huang, Xiaopei; Yang, Yiping (2010) Direct TLR2 signaling is critical for NK cell activation and function in response to vaccinia viral infection. PLoS Pathog 6:e1000811
Barral, Paola M; Sarkar, Devanand; Su, Zao-zhong et al. (2009) Functions of the cytoplasmic RNA sensors RIG-I and MDA-5: key regulators of innate immunity. Pharmacol Ther 124:219-34
Ishikawa, Hiroki; Ma, Zhe; Barber, Glen N (2009) STING regulates intracellular DNA-mediated, type I interferon-dependent innate immunity. Nature 461:788-92
Ishikawa, Hiroki; Barber, Glen N (2008) STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling. Nature 455:674-8