Biomarkers for early detection of asbestosassociated
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1 Biomarkers for early detection of asbestosassociated cancers ( study) G. Johnen, B. Pesch, D. Weber, K. Gawrych, S. Casjens, I. Raiko, O. Bryk, D. Taeger, S. Meier, P. Rozynek, T. Wiethege, G. Aguilar-Madrid, T. Brüning 3 June 2015, MEDICHEM Minisymposium, 31 st ICOH 2015
2 Follow-up exams for asbestos-exposed workers in Germany Exams are offered to:» Workers with known exposure to asbestos» Workers with recognized occupational diseases, e.g., asbestosis, plaques etc. (BK 4103) 2
3 Molecular Markers () study goals» Development of new markers for the early detection of mesothelioma and lung cancer» Validation of the markers in a prospective study (utilizing existing follow-up structures of the statutory accident insurances)» Development of affordable, fast & simple assays» Application of validated markers in follow-up exams pixabay.com 3
4 part I: Identification and verification of new markers» Detection in blood (plasma, cells, whole blood)» Different molecular levels: Protein DNA (copy number variations) Epigenetic (DNA methylation) RNA (micrornas, lncrnas)» Panel of different new markers 4
5 part II prospective study for marker validation» >2.000 Subjects» Asbestosis, plaques, etc. (BK 4103)» Yearly exams» Multicentric, 1 central lab» Logistics» Automated sample preparation» Epidemiology 5
6 Phases of biomarker development Discovery Case-control study (cross-sectional) Identification of new markers Verification Case-control study (cross-sectional) Verification on different levels: Development of robust assays - mechanistic - independ. collective - independ. method Validation Prospective cohort study (longitudinal) Biobank 6
7 Current Status of, part I (May 2015) Cross-sectional design (discovery & verification):» > 350 Cases & 700 controls» Assays for calretinin, mesothelin & mir-103 verified as marker for mesothelioma» More assays in development: DNA methylation, micrornas, proteins 7
8 Verification of Calretinin in independent cohorts: Mexico & Germany 100 Cases Controls 10 Calretinin [ng/ml] N=61 N=80 N=109 N=273 GER MEX GER MEX 8
9 Verification of mir-103 in independent cohorts: Mexico & Germany p < p = mir-103a-3p (2 -dct ) mir-103a-3p (2 -dct ) Mesothelioma Mesotheliome (N=43) Kontrollen Controls (N=52) 1 Mesothelioma Mesotheliome (N=13) Kontrollen Controls (N=13) 9
10 Performance of biomarkers in case-control studies (males only, sarcomatoid mesotheliomas excluded, preset high specificity) Marker for Mesothelioma False- Positives Sensitivity Specificity Calretinin 3 of % 99% Mesothelin 3 of % 99% mir of 13 46% 100% à high specificity allows combination of markers to increase sensitivity 10
11 Combination of biomarkers in case-control studies (males only, sarcomatoid mesotheliomas excluded) Marker combinations for Mesothelioma Sensitivity Specificity Mesothelin 55% 99% Mesothelin + Calretinin ( * ) 66% 98% Mesothelin + mir-103 ( ** ) 74% 96% ( * ) Gawrych et al., DGAUM 2015 ( ** ) Weber et al. PLoS One 2014 à Combination of markers leads to increase of sensitivity 11
12 Potential biomarkers for lung cancer (DNA methylation) LINE1 DLMS001 DLMS003 12
13 Phases of biomarker development Discovery Case-control study (cross-sectional) Identification of new markers Verification Case-control study (cross-sectional) Verification on different levels: Development of robust assays - mechanistic - independ. collective - independ. method Validation Prospective cohort study (longitudinal) Biobank 13
14 Differences between cross-sectional and longitudinal design Event cross-sectional prospective/longitudinal Clinical symptoms Cancer diagnosis Exam Blood collection Exam Blood collection Exam Exam Blood collection Blood collection Exam Blood collection Increase of marker Time Critical window: months 14
15 Current Status of, part II (May 2015) Prospective design (validation):» Workers (rate of participation: 86%)» Medical exams» 180,000 Aliquots in biobank» 25 Medical centers» 13 Mesotheliomas, 25 lung cancers 15
16 Biobank Logistics Automated aliquoting Infrastructure Backup Sample storage Database 16
17 Future research network of biobanks & cohorts Wismut Natl. Cohort As, quartz, radiation Controls Wikipedia Wikipedia Asbestos Clinical collective Controls Aromat. amines HNRECALL Asbestos,... PURE UroScreen Wikipedia LD-HRCT 17
18 Conclusions» Marker assays established & verified» Combination of markers for panel» Tools for prospective cohort established» Biobank established» Recruitment goals met Next:» First evaluation of markers in longitudinal design Future:» Use of validated markers for early detection in Mexico, Greece & Germany 18
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