MALDI imaging mass spectrometry: Discrimination of pathophysiological regions in traumatized skeletal muscle by characteristic peptide signatures

Authors

  • Oliver Klein,

    Corresponding author
    1. Berlin-Brandenburg Center for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
    2. Core Unit Proteomics, Berlin-Brandenburg Center for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
    • Correspondence: Oliver Klein, Berlin-Brandenburg Center for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Augustenburger Platz 1, 13353 Berlin, Germany

      E-mail: Oliver.klein@charite.de

      Fax: +49-30-450566904

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  • Kristin Strohschein,

    1. Julius Wolff Institute and Center for Musculoskeletal Surgery, Charité—Universitätsmedizin Berlin, Berlin, Germany
    2. Berlin-Brandenburg School for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
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  • Grit Nebrich,

    1. Berlin-Brandenburg Center for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
    2. Core Unit Proteomics, Berlin-Brandenburg Center for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
    3. Julius Wolff Institute and Center for Musculoskeletal Surgery, Charité—Universitätsmedizin Berlin, Berlin, Germany
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  • Janina Oetjen,

    1. Center for Industrial Mathematics, University of Bremen, Bremen, Germany
    2. MALDI Imaging Lab, Department of Biology and Chemistry, University of Bremen, Bremen, Germany
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  • Dennis Trede,

    1. Center for Industrial Mathematics, University of Bremen, Bremen, Germany
    2. SCiLS GmbH, Fahrenheitstr, Bremen, Germany
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  • Herbert Thiele,

    1. SCiLS GmbH, Fahrenheitstr, Bremen, Germany
    2. Fraunhofer, Institute for Medical Image Computing MEVIS, Lübeck, Germany
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  • Theodore Alexandrov,

    1. Center for Industrial Mathematics, University of Bremen, Bremen, Germany
    2. MALDI Imaging Lab, Department of Biology and Chemistry, University of Bremen, Bremen, Germany
    3. SCiLS GmbH, Fahrenheitstr, Bremen, Germany
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  • Patrick Giavalisco,

    1. Experimental Systems Biology, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany
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  • Georg N. Duda,

    1. Berlin-Brandenburg Center for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
    2. Julius Wolff Institute and Center for Musculoskeletal Surgery, Charité—Universitätsmedizin Berlin, Berlin, Germany
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  • Philipp von Roth,

    1. Julius Wolff Institute and Center for Musculoskeletal Surgery, Charité—Universitätsmedizin Berlin, Berlin, Germany
    2. Berlin-Brandenburg School for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
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  • Sven Geissler,

    1. Berlin-Brandenburg Center for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
    2. Julius Wolff Institute and Center for Musculoskeletal Surgery, Charité—Universitätsmedizin Berlin, Berlin, Germany
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  • Joachim Klose,

    1. Berlin-Brandenburg Center for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
    2. Core Unit Proteomics, Berlin-Brandenburg Center for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
    3. Institute of Human Genetics, Charité—Universitätsmedizin Berlin, Berlin, Germany
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  • Tobias Winkler

    1. Berlin-Brandenburg Center for Regenerative Therapies, Charité—Universitätsmedizin Berlin, Berlin, Germany
    2. Julius Wolff Institute and Center for Musculoskeletal Surgery, Charité—Universitätsmedizin Berlin, Berlin, Germany
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Abstract

Due to formation of fibrosis and the loss of contractile muscle tissue, severe muscle injuries often result in insufficient healing marked by a significant reduction of muscle force and motor activity. Our previous studies demonstrated that the local transplantation of mesenchymal stromal cells into an injured skeletal muscle of the rat improves the functional outcome of the healing process. Since, due to the lack of sufficient markers, the accurate discrimination of pathophysiological regions in injured skeletal muscle is inadequate, underlying mechanisms of the beneficial effects of mesenchymal stromal cell transplantation on primary trauma and trauma adjacent muscle area remain elusive. For discrimination of these pathophysiological regions, formalin-fixed injured skeletal muscle tissue was analyzed by MALDI imaging MS. By using two computational evaluation strategies, a supervised approach (ClinProTools) and unsupervised segmentation (SCiLS Lab), characteristic m/z species could be assigned to primary trauma and trauma adjacent muscle regions. Using “bottom-up” MS for protein identification and validation of results by immunohistochemistry, we could identify two proteins, skeletal muscle alpha actin and carbonic anhydrase III, which discriminate between the secondary damage on adjacent tissue and the primary traumatized muscle area. Our results underscore the high potential of MALDI imaging MS to describe the spatial characteristics of pathophysiological changes in muscle.

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