Lipid Analysis of Airway Epithelial Cells for Studying Respiratory Diseases.
Airway epithelial cells play an important role in the pathogenesis of inflammatory lung diseases such as asthma, cystic fibrosis and COPD. Studies concerning the function of the lipid metabolism of the airway epithelium are so far based only on the detection of lipids by immunohistochemistry but quantitative analyses have not been performed. Although recent advances in mass spectrometry have allowed to identify a variety of lipid classes simultaneously in isolated tissue samples, up until now, these methods were not suitable to analyze lipids in the airway epithelium. To determine all major lipid classes in airway epithelial cells, we used an LC-MS-based approach that can easily be combined with the specific isolation procedure to obtain epithelial cells. We tested the suitability of this method with a mouse model of experimental asthma. In response to allergen challenge, perturbations in the sphingolipids were detected, which led to increased levels of ceramides. We expanded the scope of this approach analysing human bronchus samples without pathological findings of adenocarcinoma patients. For the human lung epithelium an unusual lipid class distribution was found in which ceramide was the predominant sphingolipid. In summary, we show that disease progression and lipid metabolism perturbation can be monitored in animal models and that the method can be used for the analysis of clinical samples.
Division of Bioanalytical Chemistry, Research Center Borstel, Parkallee 1-40, 23845 Borstel, Germany ; Division of Cellular Microbiology, Research Center Borstel, Parkallee 1-40, 23845 Borstel, Germany ; German Center for Infection Research, TTU-Tb, Location Borstel, Parkallee 1, 23845 Borstel, Germany.
Nicole
N
Zehethofer
Institute for Anatomy, University of Lübeck, Ratzeburger Allee 160, 23562 Lübeck, Germany.
Saskia
S
Bermbach
Institute of Laboratory Medicine and Pathochemistry, Molecular Diagnostics, Philipps University of Marburg, ZTI, Hans-Meerwein-Str. 3, 35043 Marburg, Germany ; Universities of Gießen and Marburg Lung School (UGMLC), German Center for Lung Research (DZL), Gießen, Germany.
Stefanie
S
Hagner
Institute of Laboratory Medicine and Pathochemistry, Molecular Diagnostics, Philipps University of Marburg, ZTI, Hans-Meerwein-Str. 3, 35043 Marburg, Germany ; Universities of Gießen and Marburg Lung School (UGMLC), German Center for Lung Research (DZL), Gießen, Germany.
Holger
H
Garn
Division of Clinical and Experimental Pathology, Research Center Borstel, Parkallee 1-40, 23845 Borstel, Germany.
Julia
J
Müller
Division of Clinical and Experimental Pathology, Research Center Borstel, Parkallee 1-40, 23845 Borstel, Germany ; Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL), 22927 Grosshansdorf, Germany.
Torsten
T
Goldmann
Division of Bioanalytical Chemistry, Research Center Borstel, Parkallee 1-40, 23845 Borstel, Germany.
Buko
B
Lindner
Division of Bioanalytical Chemistry, Research Center Borstel, Parkallee 1-40, 23845 Borstel, Germany ; Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL), 22927 Grosshansdorf, Germany ; German Center for Infection Research, TTU-Tb, Location Borstel, Parkallee 1, 23845 Borstel, Germany.
Dominik
D
Schwudke
Institute for Anatomy, University of Lübeck, Ratzeburger Allee 160, 23562 Lübeck, Germany ; Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL), 22927 Grosshansdorf, Germany.
Peter
P
König
Ceramides; Lipidomics; Lung airway epithelium; Murine model of experimental asthma; Sphingolipids