Influenza A virus is a highly contagious virus that causes upper and lower respiratory tract infections resulting in 200,000 hospitalizations and 36,000 deaths in the United States annually, and new influenza strains generate recurring epidemics and pandemics with significant attributable morbidity and mortality. Most of the mortality associated with influenza A infection is attributable to development of the acute respiratory distress syndrome (ARDS). Acute lung injury (ALI) and ARDS are defined by damage to the alveolar epithelium and endothelium, which allows the exudation of protein-rich fluid into the alveolar space. In our preliminary data, we show that vimentin, a type III intermediate filament protein, is required for the activation of the NLRP3 inflammasome. We provide preliminary data that vimentin-/- mice are protected from lung viral pneumonia following infection with influenza A virus (IAV). Increasing evidence from our group and others suggests that these filamentous cytoskeleton structures play key roles in signal transduction pathways and provide a scaffold for the formation and activation of protein complexes, such as the NRLP3 inflammasome. We show that NLRP3 interacts with vimentin and that this protein-protein interaction is required for the processing and maturation of pro-IL- 1? into biologically active IL 1?. Additionally, we provide preliminary data showing that vimentin is required for the interaction and translocation of NOD2 to the outer mitochondrial membrane, which results in the NOD2-mediated activation of IRF3 and release of interferon- from the IAV- infected cells. Based on these preliminary data, we hypothesize that vimentin acts as scaffold for the assembly and activation of the NLRP3 inflammasome and that NOD2 protein interaction with vimentin is required for the activation of IRF3 signaling. We have formulated three interrelated specific aims to study the regulation of vimentin intermediate filaments in both in vivo and in vitro models of influenza A-induced lung injury:
Specific Aim 1 : To determine the mechanism by which vimentin contributes to activation of NLR proteins during influenza A virus-induced acute lung injury.
Specific Aim 2 : To define the protein domain(s) in vimentin required for interaction with and activation of the NLRP3 inflammasome.
Specific Aim 3 : To determine whether the interaction between vimentin and NOD2 is required for the activation of IRF3 and the release of interferon from the IAV-infected cells.
Acute lung injury (ALI) and ARDS manifest as diffuse alveolar damage, capillary injury, and exudation of protein-rich fluid into the alveolar space, and infectio with an IAV is an important cause of ARDS. We observed that vimentin is required for the assembly and activation of the NLRP3 inflammasome and NOD2 signaling pathway. The importance of this finding for the innate immune response to IAV as well as the molecular interactions that underlie the role played by vimentin in inflammasome activation will be explored in this application.
| Misharin, Alexander V; Morales-Nebreda, Luisa; Reyfman, Paul A et al. (2017) Monocyte-derived alveolar macrophages drive lung fibrosis and persist in the lung over the life span. J Exp Med 214:2387-2404 |
| Bharat, Ankit; Bhorade, Sangeeta M; Morales-Nebreda, Luisa et al. (2016) Flow Cytometry Reveals Similarities Between Lung Macrophages in Humans and Mice. Am J Respir Cell Mol Biol 54:147-9 |
| Ridge, Karen M; Shumaker, Dale; Robert, Amélie et al. (2016) Methods for Determining the Cellular Functions of Vimentin Intermediate Filaments. Methods Enzymol 568:389-426 |
| Herold, Susanne; Becker, Christin; Ridge, Karen M et al. (2015) Influenza virus-induced lung injury: pathogenesis and implications for treatment. Eur Respir J 45:1463-78 |
| Coates, Bria M; Staricha, Kelly L; Wiese, Kristin M et al. (2015) Influenza A Virus Infection, Innate Immunity, and Childhood. JAMA Pediatr 169:956-63 |
| dos Santos, Gimena; Rogel, Micah R; Baker, Margaret A et al. (2015) Vimentin regulates activation of the NLRP3 inflammasome. Nat Commun 6:6574 |
| Morales-Nebreda, Luisa; Chi, Monica; Lecuona, Emilia et al. (2014) Intratracheal administration of influenza virus is superior to intranasal administration as a model of acute lung injury. J Virol Methods 209:116-20 |
| dos Santos, Gimena; Kutuzov, Mikhail A; Ridge, Karen M (2012) The inflammasome in lung diseases. Am J Physiol Lung Cell Mol Physiol 303:L627-33 |
