Mechanical ventilation is frequently used to support critically ill patients even in the absence of overt lung injury. A recent study, however, has linked exposure to mechanical ventilation with subsequent development of the acute respiratory distress syndrome (ARDS). The mechanism linking mechanical ventilation with initiation of lung injury is unknown. We have shown that mechanical ventilation can alter the transcriptional response to low-dose bacterial product exposure leading to excessive inflammation and early development of lung injury. Our long-term goal is to identify how mechanical ventilation modulates the transcriptional response to microbial pathogens as a first step toward developing specific interventions to improve survival in mechanically ventilated patients. The specific hypothesis for this proposal is that mechanical ventilation activates the activator protein-1 (AP-1) transcription factor, which augments gene transcription in response to pathogen-associated molecular pattern (PAMP) recognition by toll-like receptors (TLRs). We base this hypothesis on the observations that: A) mechanical ventilation preferentially induces transcription of genes with a promoter region containing the AP-1 binding sequence, B) mechanical ventilation causes nuclear translocation of AP-1, C) mechanical ventilation synergistically augments mRNA levels of genes transcribed in response to lipopolysaccharide (LPS), a cell wall component of gram-negative bacteria, and D) inhibition of c-Jun amino-terminal kinase (JNK - a kinase activator of AP-1) is protective against other types of lung injury.
The specific aims of this proposal are to: 1) Determine whether ventilator augmentation of PAMP-induced inflammation is (a) specific for individual TLR- signaling pathways and/or (b) dependent on generation of endogenous TLR ligands.
This aim i s accomplished through the combination of highly specific toll-like receptor agonists and genetically modified mice, which lack either the MyD88 or TRIF adapter protein necessary for specific TLR signaling. 2) Determine the lung cell type primarily responsive to ventilator-induced immunomodulation by (a) measuring response to TLR ligands and ventilation in chimeric mice, which lack MyD88 in either myeloid or non- myeloid cells and (b) measuring response to TLR ligands and ventilation in transgenic mice in which distal lung epithelial cells have an isolated defect in downstream TLR signaling pathways. 3) Determine the role of AP-1 activation in ventilator-augmentation of LPS-induced inflammation by (a) using chromatin immunoprecipitation assay to identify which protein subunits of AP-1 are recruited to target gene promoters by mechanical ventilation, (b) inhibiting AP-1 activation during mechanical ventilation and LPS exposure, and (c) using RNA interference to determine the function of specific AP-1 subunits identified in (a). Relevance for public health - The incidence of acute lung injury (ALI) is 78.9 per 100,000 patient-years with a mortality rate of 38.5%. Based on these values, there are an estimated 190,600 cases of ALI in the U.S. each year with 74,500 associated deaths. Additionally, the economic impact of ALI is high secondary to prolonged ICU care and overall length of hospitalization and to chronic post-hospitalization disability. These data highlight the importance of further research to help understand the factors, which promote the development of ALI. This project will help identify mechanisms by which mechanical ventilation support causes lung injury;thereby, providing guidance for the development of future interventions to decrease the incidence and severity of acute lung injury.
|Gharib, Sina A; Mar, Daniel; Bomsztyk, Karol et al. (2016) SYSTEM-WIDE MAPPING OF ACTIVATED CIRCUITRY IN EXPERIMENTAL SYSTEMIC INFLAMMATORY RESPONSE SYNDROME. Shock 45:148-56|
|Grazioli, Serge; Gil, Sucheol; An, Dowon et al. (2015) CYR61 (CCN1) overexpression induces lung injury in mice. Am J Physiol Lung Cell Mol Physiol 308:L759-65|
|Bomsztyk, Karol; Mar, Daniel; An, Dowon et al. (2015) Experimental acute lung injury induces multi-organ epigenetic modifications in key angiogenic genes implicated in sepsis-associated endothelial dysfunction. Crit Care 19:225|
|Campanholle, Gabriela; Mittelsteadt, Kristen; Nakagawa, Shunsaku et al. (2013) TLR-2/TLR-4 TREM-1 signaling pathway is dispensable in inflammatory myeloid cells during sterile kidney injury. PLoS One 8:e68640|
|Gharib, Sina A; Altemeier, William A; Van Winkle, Laura S et al. (2013) Matrix metalloproteinase-7 coordinates airway epithelial injury response and differentiation of ciliated cells. Am J Respir Cell Mol Biol 48:390-6|
|Hung, Chi; Linn, Geoffrey; Chow, Yu-Hua et al. (2013) Role of lung pericytes and resident fibroblasts in the pathogenesis of pulmonary fibrosis. Am J Respir Crit Care Med 188:820-30|
|Altemeier, William A; Liles, W Conrad; Villagra-Garcia, Ana et al. (2013) Ischemia-reperfusion lung injury is attenuated in MyD88-deficient mice. PLoS One 8:e77123|
|Altemeier, William A; Schlesinger, Saundra Y; Buell, Catherine A et al. (2012) Transmembrane and extracellular domains of syndecan-1 have distinct functions in regulating lung epithelial migration and adhesion. J Biol Chem 287:34927-35|
|Bundesmann, Michael M; Wagner, Teresa E; Chow, Yu-Hua et al. (2012) Role of urokinase plasminogen activator receptor-associated protein in mouse lung. Am J Respir Cell Mol Biol 46:233-9|
|Klaff, Lindy S; Gill, Sean E; Wisse, Brent E et al. (2012) Lipopolysaccharide-induced lung injury is independent of serum vitamin D concentration. PLoS One 7:e49076|
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