Daniel Kazdal1,2, Rémi Longuespée1, Steffen Dietz2,3,4, Rita Casadonte5, Kristina Schwamborn6, Anna-Lena Volckmar1, Jörg Kriegsmann5,7, Katharina Kriegsmann8, Margaux Fresnais9,10, Albrecht Stenzinger1,4, Holger Sültmann2,3,4, Arne Warth1,2, Mark Kriegsmann1. 1. Institute of Pathology, University Hospital Heidelberg, 69120 Heidelberg, Germany. 2. Member of the Translational Lung Research Center (TLRC) Heidelberg, German Center for Lung Research (DZL), 69120 Heidelberg, Germany. 3. Cancer Genome Research Group, German Cancer Research Center (DKFZ) and, National Center for Tumor Diseases (NCT), 69120 Heidelberg, Germany. 4. Member of the German Cancer Consortium (DKTK), 69120 Heidelberg, Germany. 5. Proteopath, 54296 Trier, Germany. 6. Institute of Pathology, TU Munich, 81675 Munich, Germany. 7. MVZ for Histology, Cytology and Molecular Diagnostics Trier, 54296 Trier, Germany. 8. Department of Hematology, Oncology and Rheumatology, University of Heidelberg, 69120 Heidelberg, Germany. 9. Department of Clinical Pharmacology and Pharmacoepidemiology, University of Heidelberg, 69120 Heidelberg, Germany. 10. German Cancer Consortium (DKTK)-German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
Abstract
PURPOSE: Matrix assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-MSI) is a powerful tool to analyze the spatial distribution of peptides in tissues. Digital PCR (dPCR) is a method to reliably detect genetic mutations. Biopsy material is often limited due to minimally invasive techniques, but information on diagnosis, prognosis, and prediction is required for subsequent clinical decision making. Thus, saving tissue material during diagnostic workup is highly warranted for best patient care. The possibility to combine proteomic analysis by MALDI-MSI and mutational analysis by dPCR from the same tissue section is evaluated. EXPERIMENTAL DESIGN: Ten 0.5 × 0.5 cm formalin-fixed paraffin embedded tissue samples of pulmonary adenocarcinomas with known EGFR or KRAS mutations are analyzed by MALDI-MSI. Subsequently, DNA is extracted from the analyzed tissue material and tested for the respective driver mutation by dPCR. RESULTS: Detection of driver gene mutations after MALDI MSI analysis is successful in all analyzed samples. Determined mutant allele frequencies are in good agreement with values assessed from untreated serial tissue sections with a mean absolute deviation of 0.16. CONCLUSION AND CLINICAL RELEVANCE: It has been demonstrated that MALDI-MSI can be combined with genetic analysis, like dPCR. Workflows enabling the subsequent analysis of proteomic and genetic markers are particularly promising for the analysis of limited sample material such as biopsy specimen.
PURPOSE: Matrix assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-MSI) is a powerful tool to analyze the spatial distribution of peptides in tissues. Digital PCR (dPCR) is a method to reliably detect genetic mutations. Biopsy material is often limited due to minimally invasive techniques, but information on diagnosis, prognosis, and prediction is required for subsequent clinical decision making. Thus, saving tissue material during diagnostic workup is highly warranted for best patient care. The possibility to combine proteomic analysis by MALDI-MSI and mutational analysis by dPCR from the same tissue section is evaluated. EXPERIMENTAL DESIGN: Ten 0.5 × 0.5 cm formalin-fixed paraffin embedded tissue samples of pulmonary adenocarcinomas with known EGFR or KRAS mutations are analyzed by MALDI-MSI. Subsequently, DNA is extracted from the analyzed tissue material and tested for the respective driver mutation by dPCR. RESULTS: Detection of driver gene mutations after MALDI MSI analysis is successful in all analyzed samples. Determined mutant allele frequencies are in good agreement with values assessed from untreated serial tissue sections with a mean absolute deviation of 0.16. CONCLUSION AND CLINICAL RELEVANCE: It has been demonstrated that MALDI-MSI can be combined with genetic analysis, like dPCR. Workflows enabling the subsequent analysis of proteomic and genetic markers are particularly promising for the analysis of limited sample material such as biopsy specimen.
Authors: Julia Hess; Kristian Unger; Lisa Kreutzer; Peter Weber; Theresa Heider; Mathias Heikenwälder; Tobias Riedl; Philipp Baumeister; Frederick Klauschen; Claus Belka; Axel Walch; Horst Zitzelsberger Journal: Lab Invest Date: 2022-08-31 Impact factor: 5.502
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Authors: Linfeng Xu; Kai-Chun Chang; Emory M Payne; Cyrus Modavi; Leqian Liu; Claire M Palmer; Nannan Tao; Hal S Alper; Robert T Kennedy; Dale S Cornett; Adam R Abate Journal: Nat Commun Date: 2021-11-23 Impact factor: 14.919