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Characterization of vestibular schwannoma tissues using liquid chromatography-tandem mass spectrometry analysis of specific peptide fragments separated by in-sample tryptic protein digestion followed by mathematical analysis
Authors:Michaela Tesařová  Tereza Boušková  Pavel Cejnar  Jiří Šantrůček  Lenka Peterková  Zdeněk Fík  Petra Sázelová  Václav Kašička  Radovan Hynek
Institution:1. Department of Otorhinolaryngology and Head and Neck Surgery, First Faculty of Medicine Charles University and Motol University Hospital, Prague 5, Czech Republic

Institute of Anatomy, First Faculty of Medicine, Charles University, Prague 2, Czech Republic;2. Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague 6, Czech Republic;3. Department of Mathematics, Informatics and Cybernetics, University of Chemistry and Technology, Prague 6, Czech Republic;4. Department of Otorhinolaryngology and Head and Neck Surgery, First Faculty of Medicine Charles University and Motol University Hospital, Prague 5, Czech Republic;5. Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague 6, Czech Republic

Abstract:Vestibular schwannoma is the most common benign neoplasm of the cerebellopontine angle. Its first symptoms include hearing loss, tinnitus, and vestibular symptoms, followed by cerebellar and brainstem symptoms, along with palsy of the adjacent cranial nerves. However, the clinical picture has unpredictable dynamics and currently, there are no reliable predictors of tumor behavior. Hence, it is desirable to have a fast routine method for analysis of vestibular schwannoma tissues at the molecular level. The major objective of this study was to verify whether a technique using in-sample specific protein digestion with trypsin would have the potential to provide a proteomic characterization of these pathological tissues. The achieved results showed that the use of this approach with subsequent liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of released peptides allowed a fast identification of a considerable number of proteins in two differential parts of vestibular schwannoma tissue as well as in tissues of control healthy samples. Furthermore, mathematical analysis of MS data was able to discriminate between pathological vestibular schwannoma tissues and healthy tissues. Thus, in-sample protein digestion combined with LC-MS/MS separation and identification of released specific peptides followed by mathematical analysis appears to have the potential for routine characterization of vestibular schwannomas at the molecular level. Data are available via ProteomeXchange with identifier PXD045261.
Keywords:In-sample protein digestion  liquid chromatography  principal component analysis  tandem mass spectrometry  vestibular schwannomas
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