Educational Immersion Experience Title: Role of Inflammatory Airway Disease in Cystic Fibrosis
Mentors: Carolyn Cannon, Ph.D., M.D., and Thomas Ferkol, M.D.
Location: MPRB 6th floor laboratory
Overall Educational Goals: Despite recent advances, cystic fibrosis (CF) remains one of the most
common genetic killers in the United States. In CF, defective function of the cystic fibrosis
transmembrane conductance regulator (CFTR) in airway epithelial cells and submucosal glands results in
the chronic involvement of the respiratory tract, manifested by airway obstruction and recurrent infections
of the lung and paranasal sinuses that begins early in life. Although infection contributes to the morbidity
of patients with CF, the intense host inflammatory response accounts for the progressive, suppurative
pulmonary disease, which ultimately leads to death. Several lines of evidence suggest that the
inflammatory response is excessive to the threat posed by infection, and that relatively minor infections
induce a robust inflammatory reaction in the CF lung, which does not subside. Moreover, it has become
increasingly clear that CF exists as a continuum, and clinical manifestations of the disease are
dependent on the presence of mutant CFTR allele(s), altered by genetic modifiers and environmental
factors.
The primary goal of this rotation is to provide the trainee with experience and insights into the use of in
vitro respiratory epithelial cell models to understand inflammatory responses of airway epithelia to various
stimuli and to identify the genetic and molecular defects of CF that leads to an exaggerated inflammatory
response in the lung.
Educational goals by week:
Weeks 1-4: To gain familiarity with and experience in respiratory epithelial cell culture techniques,
including the collection and maintenance of murine airway epithelial cells, as well as the molecular and
immunoassys used to measure inflammatory responses.
Weeks 4-8: To learn how to create polarized, murine airway epithelial cell models, grown in primary
culture at an air-liquid interface, which permits evaluation of inflammatory gene regulation and expression
in mouse respiratory epithelia.
Weeks 8-12: To compare the inflammatory phenotypes and chemokine expression patterns of wildtype
and cystic fibrosis airway epithelial cells in response to stimulation.
Suggested readings:
Knowles MR, Boucher RC. Mucus clearance as a primary innate defense mechanism for mammalian
airways. J Clin Invest 2002;109:571-577.
van Heeckeren A, Walenga R, Konstan MW, Bonfield T, Davis PB, Ferkol T. Excessive inflammatory
response of cystic fibrosis mice to bronchopulmonary infection with Pseudomonas aeruginosa. J Clin Invest
1997;100:2810-2815.
Aldallal N, McNaughton EE, Manzel LJ, Rabe AM, Karp PH, Zabner J, Jacquot J, Ferkol TW, Look DC.
Inflammatory response in airway epithelial cells isolated from cystic fibrosis lungs. Am J Respir Crit Care
Med 2002;166:1248-56.
Joseph T, Look DC, Ferkol TW. NF-B activation and sustained IL-8 gene expression in primary cultures of
cystic fibrosis airway epithelial cells stimulated with Pseudomonas aeruginosa. Am J Physiol, Lung Cell Mol
Physiol. 2005;288:L471-9.