Label-free single-cell phenotyping to determine tumor cell heterogeneity in pancreatic cancer in real time.

Journal: JCI insight
Published Date: Jul 8, 2025

Abstract

Resistance to chemotherapy of pancreatic ductal adenocarcinoma (PDAC) is largely driven by intratumoral heterogeneity (ITH) due to tumor cell plasticity and clonal diversity. To develop alternative strategies to overcome this defined mechanism of resistance, tools to monitor and quantify ITH in a rapid and scalable fashion are needed urgently. Here, we employed label-free digital holographic microscopy (DHM) to characterize ITH in PDAC. We established a robust experimental and machine learning analysis pipeline to perform single-cell phenotyping based on DHM-derived phase images of PDAC cells in suspension. Importantly, we were able to detect dynamic changes in tumor cell differentiation and heterogeneity of distinct PDAC subtypes upon induction of epithelial-mesenchymal transition and under treatment-imposed pressure in murine and patient-derived model systems. This platform allowed us to assess phenotypic ITH in PDAC on a single-cell level in real time. Implementing this technology into the clinical workflow has the potential to fundamentally increase our understanding of tumor heterogeneity during evolution and treatment response.

Authors

  • Katja Wittenzellner
    Translational Pancreatic Cancer Research Center, TUM School of Medicine and, University Medical Center, Technical University of Munich, Munich, Germany.
  • Manuel Lengl
    Chair for Data Processing, Technical University of Munich, Munich, Germany.
  • Stefan Röhrl
    Chair for Data Processing, Technical University of Munich, Munich, Germany.
  • Carlo Maurer
    Translational Pancreatic Cancer Research Center, TUM School of Medicine and, University Medical Center, Technical University of Munich, Munich, Germany.
  • Christian Klenk
    Heinz Nixdorf Chair of Biomedical Electronics, TranslaTUM, Technical University of Munich, Munich, Germany.
  • Aristeidis Papargyriou
    Translational Pancreatic Cancer Research Center, TUM School of Medicine and, University Medical Center, Technical University of Munich, Munich, Germany.
  • Laura Schmidleitner
    Translational Pancreatic Cancer Research Center, TUM School of Medicine and, University Medical Center, Technical University of Munich, Munich, Germany.
  • Nicole Kabella
    Chair of Proteomics and Bioanalytics, Technical University of Munich, Freising, Germany.
  • Akul Shastri
    Translational Pancreatic Cancer Research Center, TUM School of Medicine and, University Medical Center, Technical University of Munich, Munich, Germany.
  • David E Fresacher
    Chair for Data Processing, Technical University of Munich, Munich, Germany.
  • Farid Harb
    Chair for Data Processing, Technical University of Munich, Munich, Germany.
  • Nawal Hafez
    Chair for Data Processing, Technical University of Munich, Munich, Germany.
  • Stefanie Bärthel
    Institute for Translational Cancer Research and Experimental Cancer Therapy, TranslaTUM, Technical University of Munich, Munich, Germany.
  • Daniele Lucarelli
    Institute for Translational Cancer Research and Experimental Cancer Therapy, TranslaTUM, Technical University of Munich, Munich, Germany.
  • Carmen Escorial-Iriarte
    Translational Pancreatic Cancer Research Center, TUM School of Medicine and, University Medical Center, Technical University of Munich, Munich, Germany.
  • Felix Orben
    Translational Pancreatic Cancer Research Center, TUM School of Medicine and, University Medical Center, Technical University of Munich, Munich, Germany.
  • Rupert Öllinger
    Institute of Molecular Oncology and Functional Genomics, TranslaTUM, TUM School of Medicine, Technical University of Munich, Munich, Germany.
  • Ellen Emken
    Heinz Nixdorf Chair of Biomedical Electronics, TranslaTUM, Technical University of Munich, Munich, Germany.
  • Lisa Fricke
    Translational Pancreatic Cancer Research Center, TUM School of Medicine and, University Medical Center, Technical University of Munich, Munich, Germany.
  • Joanna Madej
    Translational Pancreatic Cancer Research Center, TUM School of Medicine and, University Medical Center, Technical University of Munich, Munich, Germany.
  • Patrick Wustrow
    Translational Pancreatic Cancer Research Center, TUM School of Medicine and, University Medical Center, Technical University of Munich, Munich, Germany.
  • I Ekin Demir
    Department of Surgery, TUM Universitätsklinikum, Klinikum rechts der Isar, Technical University of Munich, Munich, Germany.
  • Helmut Friess
    Department of Surgery, School of Medicine, Technical University of Munich, Munich, Germany.
  • Tobias Lahmer
    Klinik Und Poliklinik Für Innere Medizin II, Klinikum Rechts Der Isar der, Technischen Universität München, Munich, Germany.
  • Roland M Schmid
    II. Medizinische Klinik und Poliklinik. Klinikum rechts der Isar der Technischen Universität München, 81675 München, Germany.
  • Roland Rad
    Center for Translational Cancer Research (TranslaTUM), TUM School of Medicine, Technical University of Munich (TUM), Munich, Germany.
  • Günter Schneider
    Department of General, Visceral and Pediatric Surgery, University Medical Center Göttingen, Göttingen, Germany.
  • Bernhard Kuster
    Chair for Proteomics and Bioanalytics, TU Muenchen, Freising 85354, Germany; German Cancer Consortium (DKTK), Munich, Germany; German Cancer Research Center (DKFZ), Heidelberg, Germany; Center for Integrated Protein Science Munich, Munich, Germany; Bavarian Biomolecular Mass Spectrometry Center, Technische Universität München, Freising, Germany.
  • Dieter Saur
    Center for Translational Cancer Research (TranslaTUM), Klinikum rechts der Isar, Technical University of Munich, 81675 Munich, Germany; Division of Translational Cancer Research, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; German Cancer Consortium (DKTK), Partner Site Munich, Klinikum rechts der Isar, Technische Universität München, 81675 Munich, Germany.
  • Oliver Hayden
    Heinz Nixdorf Chair of Biomedical Electronics, TranslaTUM, Technical University of Munich, Munich, Germany.
  • Klaus Diepold
    Department of Electrical and Computer Engineering, Technische Universität München, Munich, Bavaria, Germany.
  • Maximilian Reichert

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