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Abstract No.: MoP-215
Session: Proteomics: Clinical
Presentation date: Mon, Aug 28, 2006
Presentation time: 09:50 – 11:20

A Proteomic Analysis to Find Biomarkers for Early Detection of Asbestos-Related Lung Diseases

Anne Puustinen1, Niina Ahonen1, Sisko Anttila1, Matti S. Huuskonen1, Tuula Toljamo2, Katri Vuopala2, Tuula A. Nyman1

1 Finnish Institute of Occupational Health, Helsinki, Finland
2 Central Hospital of Lapland, Rovaniemi, Finland

Correspondence address: Anne Puustinen, Finnish Institute of Occupational Health, Centre of Expertise for Health and Work Ability, Topeliuksenkatu 41aA, Helsinki, 00250 Finland.

Keywords: Biomarkers; MS/MS, Liquid Chromatography; Plasma, Human; Proteomic.

Novel aspect: Discovery of possible biomarkers for early detection and treatment asbestos-related lung diseases.

 

The inhalation of asbestos fibers has been linked to the development of malignant and fibrotic diseases of the lung and pleura. Although exposure to asbestos fibers may have occurred dozens of years earlier, the long latency period of the diseases implies that many workers are still at risk of developing asbestosis and pulmonary cancer in the upcoming years. Thus, these diseases are currently and will remain a significant health problem, especially in developing countries, where the use of asbestos continues to increase.
To develop tools for early detection of the initiation of the disease process caused by inhalation of asbestos fibers, we have used a comparative plasma proteome analysis. Plasma samples were from earlier health survey studies of Finnish tobacco smoking and fiber exposed populations, and were divided into four groups of 5-6 individuals on the basis of sex, age and smoking history: former smokers, former smokers with lung cancer, former smokers with lung cancer and occupational exposure to asbestos, and former smokers with pulmonary fibrosis caused by asbestos fibers.
Because the major plasma protein components, e.g. albumin and immunoglobins, interfere with the detection of less abundant proteins, EDTA-anticoagulated plasma was depleted of the two most abundant proteins (PROTIA, Sigma) to facilitate a higher sample load for improved visualization of low copy number proteins on two-dimensional electrophoresis gels. After depletion, the remaining plasma proteins were resolved by 2-DE using three different pH interval IPG strips (pH 3-6, 5-8 and 7-10) in the first dimension, and 12% SDS-PAGE gels in the second dimension. After 2-DE, the proteins were visualised by silver-staining. To find differentially expressed protein spots, gels were analysed with Image Master Platinum software (GE Healthcare). For identification, the protein spots of interest were cut out from the gel, in-gel digested, and the resulting peptides were analysed with nanoLC-MS/MS (CapLC and Q-TOF Global, Waters Micromass), followed by database searching using Mascot (Matrix Science). Validation of the found differences is in progress.
Protein markers indentified by this approach may help in minimizing morbidity related to asbestos fiber exposure. The found alterations in protein expression could also enhance our understanding of largely unknown cellular responses to asbestos exposure.