title: Spatial and molecular resolution of diffuse malignant mesothelioma heterogeneity by integrating label-free FTIR imaging, laser capture microdissection and proteomics.
abstract: Diffuse malignant mesothelioma (DMM) is a heterogeneous malignant neoplasia manifesting with three subtypes: epithelioid, sarcomatoid and biphasic. DMM exhibit a high degree of spatial heterogeneity that complicates a thorough understanding of the underlying different molecular processes in each subtype. We present a novel approach to spatially resolve the heterogeneity of a tumour in a label-free manner by integrating FTIR imaging and laser capture microdissection (LCM). Subsequent proteome analysis of the dissected homogenous samples provides in addition molecular resolution. FTIR imaging resolves tumour subtypes within tissue thin-sections in an automated and label-free manner with accuracy of about 85% for DMM subtypes. Even in highly heterogeneous tissue structures, our label-free approach can identify small regions of interest, which can be dissected as homogeneous samples using LCM. Subsequent proteome analysis provides a location specific molecular characterization. Applied to DMM subtypes, we identify 142 differentially expressed proteins, including five protein biomarkers commonly used in DMM immunohistochemistry panels. Thus, FTIR imaging resolves not only morphological alteration within tissue but it resolves even alterations at the level of single proteins in tumour subtypes. Our fully automated workflow FTIR-guided LCM opens new avenues collecting homogeneous samples for precise and predictive biomarkers from omics studies.
authors.affiliation: Ruhr-University Bochum, Department of Biophysics, Bochum, Germany.
authors.forename: Frederik
authors.initials: F
authors.lastname: Großerueschkamp
authors.affiliation: Ruhr-University Bochum, Medizinisches Proteom- Center (MPC), Bochum, Germany.
authors.forename: Thilo
authors.initials: T
authors.lastname: Bracht
authors.affiliation: Ruhr-University Bochum, Medizinisches Proteom- Center (MPC), Bochum, Germany.
authors.forename: Hanna C
authors.initials: HC
authors.lastname: Diehl
authors.affiliation: Ruhr-University Bochum, Department of Biophysics, Bochum, Germany.
authors.forename: Claus
authors.initials: C
authors.lastname: Kuepper
authors.affiliation: Ruhr-University Bochum, Medizinisches Proteom- Center (MPC), Bochum, Germany.
authors.forename: Maike
authors.initials: M
authors.lastname: Ahrens
authors.affiliation: Ruhr-University Bochum, Department of Biophysics, Bochum, Germany.
authors.forename: Angela
authors.initials: A
authors.lastname: Kallenbach-Thieltges
authors.affiliation: Ruhr-University Bochum, Department of Biophysics, Bochum, Germany.
authors.forename: Axel
authors.initials: A
authors.lastname: Mosig
authors.affiliation: Ruhr-University Bochum, Medizinisches Proteom- Center (MPC), Bochum, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Eisenacher
authors.affiliation: Ruhr-University Bochum, Medizinisches Proteom- Center (MPC), Bochum, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Marcus
authors.affiliation: Institute for Prevention and Occupational Medicine of the German Social Accident Insurance, Institute of the Ruhr Universität Bochum (IPA), Bochum, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Behrens
authors.affiliation: Institute for Prevention and Occupational Medicine of the German Social Accident Insurance, Institute of the Ruhr Universität Bochum (IPA), Bochum, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Brüning
authors.affiliation: University Duisburg Essen, University Hospital Essen, Institute of Pathology, Essen, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Theegarten
authors.affiliation: Ruhr-University Bochum, Medizinisches Proteom- Center (MPC), Bochum, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Sitek
authors.affiliation: Ruhr-University Bochum, Department of Biophysics, Bochum, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Gerwert
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers; D006801:Humans; D007150:Immunohistochemistry; D061346:Laser Capture Microdissection; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000378:metabolism*; D008297:Male; D008654:Mesothelioma / Q000175:diagnosis* / Q000378:metabolism*; D000086002:Mesothelioma, Malignant; D008875:Middle Aged; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D017550:Spectroscopy, Fourier Transform Infrared; D057188:Workflow
chemical_list: D015415:Biomarkers; D020543:Proteome