The innate immune system senses non-self nucleic acids via germ-line encoded pattern recognition receptors. RNA is sensed via Toll-like receptor (TLR)-3, -7 and -8 or by the cytoplasmic RNA helicases RIG-I and MDA-5. Little is known about sensors for cytoplasmic DNA, which when delivered to the cytoplasm trigger type I IFN gene transcription and caspase-1-mediated processing of IL-12. We have identified PISA (PYHIN protein stimulating ASC), one of four human PYHIN proteins as a receptor for cytosolic DNA. The HIN200 domain of PISA binds to DNA, while the PYD domain (but not that of PYHIN1-3) associates with the adapter molecule ASC to activate NF-:B and caspase-1. Knockdown of PISA downregulates caspase-1-mediated IL-12 responses, with concomitant increases in type I interferon production. Our hypothesis is that PISA serves to recognize microbial cytosolic DNA (such as bacterial or viral DNA) and that PISA induces pro-inflammatory responses towards cytoplasmic DNA via the assembly of an inflammasome that engages ASC and caspase-1. In this study, we propose to further characterize the role of PISA in the recognition of viral and bacterial cytosolic DNA. We will also generate a PISA knock-out mouse by gene-trap technology and/or targeted gene deletion and propose to analyze the in vivo relevance of PISA for microbial infections. Finally, we will assess the role of PISA for the modulation of interferon responses towards cytoplasmic DNA. Elucidation of the molecular mechanisms of cytosolic DNA recognition could lead to novel therapeutic targets for microbial infections and thereby enable novel strategies that could prevent inflammatory reactions to gene therapy approaches. Public Health Relevance: Pathogens are detected by receptors expressed in different areas of cells of the immune system. This grant is focused on understanding how a cytosolic protein, which we have identified called PISA (PYHIN protein stimulating ASC) senses pathogens by virtue of their DNA. Elucidation of molecular details of PISA activation could lead to novel strategies of pharmacological interference for diseases based on intracellular DNA recognition.

Public Health Relevance

Pathogens are detected by receptors expressed in different areas of cells of the immune system. This grant is focused on understanding how a cytosolic protein, which we have identified called PISA (PYHIN protein stimulating ASC) senses pathogens by virtue of their DNA. Elucidation of molecular details of PISA activation could lead to novel strategies of pharmacological interference for diseases based on intracellular DNA recognition.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI083713-04
Application #
8291224
Study Section
Immunity and Host Defense Study Section (IHD)
Program Officer
Palker, Thomas J
Project Start
2009-07-01
Project End
2014-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
4
Fiscal Year
2012
Total Cost
$403,066
Indirect Cost
$158,041
Name
University of Massachusetts Medical School Worcester
Department
None
Type
Organized Research Units
DUNS #
603847393
City
Worcester
State
MA
Country
United States
Zip Code
01655
Weng, Dan; Marty-Roix, Robyn; Ganesan, Sandhya et al. (2014) Caspase-8 and RIP kinases regulate bacteria-induced innate immune responses and cell death. Proc Natl Acad Sci U S A 111:7391-6
Thompson, Mikayla R; Sharma, Shruti; Atianand, Maninjay et al. (2014) Interferon ?-inducible protein (IFI) 16 transcriptionally regulates type i interferons and other interferon-stimulated genes and controls the interferon response to both DNA and RNA viruses. J Biol Chem 289:23568-81
Franklin, Bernardo S; Bossaller, Lukas; De Nardo, Dominic et al. (2014) The adaptor ASC has extracellular and 'prionoid' activities that propagate inflammation. Nat Immunol 15:727-37
Kailasan Vanaja, Sivapriya; Rathinam, Vijay A K; Atianand, Maninjay K et al. (2014) Bacterial RNA:DNA hybrids are activators of the NLRP3 inflammasome. Proc Natl Acad Sci U S A 111:7765-70
Collins, James W; Keeney, Kristie M; Crepin, Valerie F et al. (2014) Citrobacter rodentium: infection, inflammation and the microbiota. Nat Rev Microbiol 12:612-23
Mangan, Matthew S J; Latz, Eicke (2014) NLRC3 puts the brakes on STING. Immunity 40:305-6
Ganesan, Sandhya; Rathinam, Vijay A K; Bossaller, Lukas et al. (2014) Caspase-8 modulates dectin-1 and complement receptor 3-driven IL-1? production in response to ?-glucans and the fungal pathogen, Candida albicans. J Immunol 193:2519-30
Kalantari, Parisa; DeOliveira, Rosane B; Chan, Jennie et al. (2014) Dual engagement of the NLRP3 and AIM2 inflammasomes by plasmodium-derived hemozoin and DNA during malaria. Cell Rep 6:196-210
Lu, Alvin; Magupalli, Venkat Giri; Ruan, Jianbin et al. (2014) Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. Cell 156:1193-206
Shah, Swati; Bohsali, Amro; Ahlbrand, Sarah E et al. (2013) Cutting edge: Mycobacterium tuberculosis but not nonvirulent mycobacteria inhibits IFN-* and AIM2 inflammasome-dependent IL-1* production via its ESX-1 secretion system. J Immunol 191:3514-8

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