Cystic fibrosis pulmonary disease is characterized by neutrophil-dominated airway inflammation, which eventually leads to fibrosis, bronchiectasis, and pulmonary failure. Pseudomonas aeruginosa infection is an important cause of this inflammatory process. In the experiments proposed, exactly how this organism stimulates inflammation in both normal and epithelial cells with mutant CFTR alleles will be delineated. Adherent P. aeruginosa stimulate epithelial cells to express large amounts of the neutrophil chemokine IL-8. This is accomplished through a complex signaling cascade. One of the earliest responses of the epithelial cell to P. aeruginosa is a brisk rise in intracellular calcium, which is followed by activation of the transcription factor of IL-9 and other pro-inflammatory cytokines. We will establish which bacterial gene products trigger the epithelial flux in calcium which is required to evoke this inflammatory response. The nature of the epithelial receptor, the kinases and phosphatases activated by the bacteria, and the regulation of this signaling pathway will be established. Endogenous stimuli such as the accumulation of mutant CFTR within the endoplasmic reticulum (ER overload) may also trigger a rise in intracellular calcium which similarly initiates activation of NF-kappaB By comparing the response of cells with specific types of CFTR mutations, it should be possible to determine how CFTR dysfunction affects the immune function of epithelial cells, as well as the electrophysiological properties. Analysis of the activation of NF-kapapB in cells with defined CFTR mutations from both transgenic mice, as well as from patients with CF may suggest therapeutic strategies to modulate the inflammatory response. Immunosuppressant drugs such as FK506 and cyclosporine A block the calcium dependent phosphatase calcineurin. It may be possible to use such drugs to diminish the inflammation provoke by P. aeruginosa as well as the endogenous stimulation of NF-kappaB produced in response to the cell stress initiated by the accumulation of mutant, mistrafficked CFTR in the endoplasmic reticulum.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Specialized Center (P50)
Project #
5P50HL060293-03
Application #
6354743
Study Section
Project Start
2000-09-01
Project End
2001-08-31
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
3
Fiscal Year
2000
Total Cost
$167,271
Indirect Cost
Name
Case Western Reserve University
Department
Type
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
van Heeckeren, Anna M; Schluchter, Mark D; Xue, Wei et al. (2006) Response to acute lung infection with mucoid Pseudomonas aeruginosa in cystic fibrosis mice. Am J Respir Crit Care Med 173:288-96
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Waters, Valerie; Sokol, Sach; Reddy, Bharat et al. (2005) The effect of cyclosporin A on airway cell proinflammatory signaling and pneumonia. Am J Respir Cell Mol Biol 33:138-44
Van Heeckeren, Anna M; Scaria, Abraham; Schluchter, Mark D et al. (2004) Delivery of CFTR by adenoviral vector to cystic fibrosis mouse lung in a model of chronic Pseudomonas aeruginosa lung infection. Am J Physiol Lung Cell Mol Physiol 286:L717-26
Muir, Amanda; Soong, Grace; Sokol, Sach et al. (2004) Toll-like receptors in normal and cystic fibrosis airway epithelial cells. Am J Respir Cell Mol Biol 30:777-83
Gupta, Sanhita; Xie, Junxia; Ma, Jianjie et al. (2004) Intermolecular interaction between R domains of cystic fibrosis transmembrane conductance regulator. Am J Respir Cell Mol Biol 30:242-8
Adamo, Robert; Sokol, Sach; Soong, Grace et al. (2004) Pseudomonas aeruginosa flagella activate airway epithelial cells through asialoGM1 and toll-like receptor 2 as well as toll-like receptor 5. Am J Respir Cell Mol Biol 30:627-34
van Heeckeren, Anna M; Schluchter, Mark; Xue, Lintong et al. (2004) Nutritional effects on host response to lung infections with mucoid Pseudomonas aeruginosa in mice. Infect Immun 72:1479-86
van Heeckeren, Anna M; Schluchter, Mark D; Drumm, Mitchell L et al. (2004) Role of Cftr genotype in the response to chronic Pseudomonas aeruginosa lung infection in mice. Am J Physiol Lung Cell Mol Physiol 287:L944-52
Ferkol, Thomas; Cohn, Leah A; Phillips, Thomas E et al. (2003) Targeted delivery of antiprotease to the epithelial surface of human tracheal xenografts. Am J Respir Crit Care Med 167:1374-9

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