Title: Proteomics in the search for (early) biomarkers of asthma
Authors: Haenen, Steven ×
Vanoirbeek, Jeroen
Hoet, Peter
Schoofs, Liliane
Clynen, Elke #
Issue Date: 2008
Conference: DIGE Users day GE Healthcare location:Eindhoven, The Netherlands date:15 October 2008
Abstract: Proteomics in the search for (early) biomarkers of asthma

Steven Haenen1,2, Jeroen A. Vanoirbeek1, Peter H. Hoet1, Liliane Schoofs2, Elke Clynen2

1Laboratory of Pneumology (Lung Toxicology), K.U. Leuven, O&N1 Herestraat 49, 3000 Leuven, Belgium
2Research Group Functional Genomics and Proteomics, K.U. Leuven, Naamsestraat 59, 3000 Leuven, Belgium

Asthma is one of the most prevalent chronic airway diseases, characterized by reversible airway obstruction, airway inflammation, and non-specific airway hyperreactivity. Up to 15% of all asthma cases in adults are due to chemical exposures at the workplace. Initially, the exposed workers do not show asthmatic symptoms but are getting sensitized. The identification of biomarkers of sensitization could help to move diagnosis to an earlier stage. Using a mouse model of chemical-induced asthma we will explore the process of sensitization towards chemicals at the protein level.

On day 1 and 8, mice are treated on both ears with 0,3% toluene diisocyanate (TDI), a known respiratory sensitizer, 3% TDI or a mixture of acetone – olive oil (vehicle-control). On day 15 mice are sacrificed and auricular lymph nodes are collected and prepared for proteome analysis.
In a second study, mice are sensitized with 0,3% TDI or a mixture of acetone – olive oil on day 1 and 8. On day 15, mice are oropharyngeally challenged with 0,01% TDI or acetone – olive oil. One day later, on day 16, lung function measurements are carried out to select responders from non-responders. The auricular lymph nodes of selected mice are subjected to proteome analysis .
In both studies two-dimensional differential gel electrophoresis (2D-DIGE) was used to analyze the differential expression of proteins in the auricular lymph nodes of TDI (0,3% or 3%) and vehicle-treated (control) mice.

In the first study, three conditions were compared in one 2D-DIGE analysis. In total, 35, 36 and 12 proteins were found differentially expressed between 0,3% TDI and control, 3% TDI and control and 0,3% TDI and 3% TDI respectively. Most proteins were significantly (p<0,05) downregulated in TDI treated mice.
In the second study, (non-)responding mice were pre-selected based on lung function measurements. 36 proteins were found to be significantly (p < 0,01) upregulated in 0,3% TDI treated mice (clear responders) and 18 proteins were downregulated. 21 proteins could already be identified by peptide mass fingerprinting. Further studies will have to point out whether (some of) these proteins could act as valuable biomarkers. Therefore, more samples (bronchoalveolar lavage fluid, serum,…) will be analysed.
Publication status: accepted
KU Leuven publication type: AMa
Appears in Collections:Occupational, Environmental and Insurance Medicine (-)
Environment and Health - miscellaneous
Animal Physiology and Neurobiology Section - miscellaneous
× corresponding author
# (joint) last author

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