High-resolution multimodal imaging reveals spatial and temporal heterogeneity of airway mucus plugging in mice with muco-obstructive lung disease

Scientific Reports, vol. 15, pp. 41760

Abstract

Mucus plugging is a hallmark of muco-obstructive lung diseases; however, the spatiotemporal dynamics of plug formation remain poorly understood. We used a multimodal high-resolution imaging approach to study mucus plugging in newborn (0-1 days), juvenile (13-16 days), and adult (49-59 days) βENaC-transgenic mice (βENaC-tg, n = 24), a model of muco-obstructive lung disease, in comparison to age-matched wild-type mice (n = 28). Micro-computed tomography (µCT) enabled whole lung mucus scoring on a scale from 0 (no obstruction) to 2 (> 50% obstruction) using an extended murine airway tree nomenclature. A subset of βENaC-tg lungs (5 juvenile, 5 adult) underwent localised synchrotron radiation-based computed tomography (SRCT) for segmentation-based calculation of mucus area and contact ratios as indicators of luminal obstruction and airway wall adherence. µCT-guided scanning electron microscopy (SEM) provided nanoscale visualisation of mucus plugs on selected βENaC-tg lung samples (3 juvenile). µCT revealed age-dependent, heterogeneous mucus plug formation, with significantly increased mucus scores in juvenile (0.45 ± 0.26) and adult (0.30 ± 0.17) βENaC-tg compared to wild-type mice (0.01 ± 0.02 with P < 0.01, and 0.01 ± 0.02 with P < 0.05, respectively). SRCT analysis showed a strong positive correlation between mucus area and contact ratios (r ≥ 0.80), while SEM revealed the mucus plug ultrastructure. This multimodal imaging approach highlights the spatiotemporal heterogeneity of mucus plugging, forming a basis for future research on targeted therapeutic strategies for muco-obstructive lung diseases.

Authors 13

1. Claudia Victoria Benke corresponding

   Heidelberg University · University Hospital Heidelberg · German Center for Lung Research

   Affiliation as printed

   Department of Diagnostic and Interventional Radiology (DIR), Heidelberg University Hospital, Heidelberg, Germany. Claudia.Benke@med.uni-heidelberg.de

   Translational Lung Research Center (TLRC), German Center for Lung Research (DZL), University Heidelberg, Heidelberg, Germany. Claudia.Benke@med.uni-heidelberg.de

   Department of Diagnostic and Interventional Radiology (DIR), Heidelberg University Hospital, Heidelberg, Germany

   Translational Lung Research Center (TLRC), German Center for Lung Research (DZL), University Heidelberg, Heidelberg, Germany

2. Julia Duerr

   Humboldt-Universität zu Berlin · German Center for Lung Research · Freie Universität Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Pediatric Respiratory Medicine, Immunology and Critical Care Medicine, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität, Berlin, Germany

   German Center for Child and Adolescent Health (DZKJ), Partner Site Berlin, Berlin, Germany

   German Center for Lung Research (DZL), Associated Partner Site Berlin, Berlin, Germany

3. Annika Engel

   University of Kaiserslautern · Universität Hamburg · Rheinland-Pfälzische Technische Universität Kaiserslautern-Landau

   Affiliation as printed

   Department of Mathematics, Technical University of Kaiserslautern (now part of RPTU University Kaiserslautern-Landau), Kaiserslautern, Germany

   Hamburg University, Hamburg, Germany

4. Christian Dullin

   Heidelberg University · University Hospital Heidelberg · Universitätsmedizin Göttingen · German Center for Lung Research · Max Planck Institute for Multidisciplinary Sciences

   Affiliation as printed

   Department of Diagnostic and Interventional Radiology (DIR), Heidelberg University Hospital, Heidelberg, Germany

   Department of Diagnostic and Interventional Radiology, University Medical Center Goettingen, Goettingen, Germany

   Translational Lung Research Center (TLRC), German Center for Lung Research (DZL), University Heidelberg, Heidelberg, Germany

   Translational Molecular Imaging, Max-Plank-Institute for Multidisciplinary Sciences, Goettingen, Germany

5. Wolfram Stiller

   Heidelberg University · University Hospital Heidelberg · German Center for Lung Research

   Affiliation as printed

   Department of Diagnostic and Interventional Radiology (DIR), Heidelberg University Hospital, Heidelberg, Germany

   Translational Lung Research Center (TLRC), German Center for Lung Research (DZL), University Heidelberg, Heidelberg, Germany

6. Heinz Horstmann

   Heidelberg University

   Affiliation as printed

   Department of Functional Neuroanatomy, Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany

7. Claudia Redenbach

   Rheinland-Pfälzische Technische Universität Kaiserslautern-Landau

   Affiliation as printed

   Department of Mathematics, RPTU University Kaiserslautern-Landau, Kaiserslautern, Germany

8. Maximilian Ackermann Aachen Institute of Pathology, University Clinics

   Witten/Herdecke University · Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz · Helios Universitätsklinikum Wuppertal · Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Anatomy, University Medical Center of the Johannes Gutenberg-University, Mainz, Germany

   Institute of Pathology and Molecular Pathology, Helios University Clinic Wuppertal, University of Witten/Herdecke, Witten, Germany

   Institute of Pathology, University Clinics of RWTH University, Aachen, Germany

9. Hans‐Ulrich Kauczor

   Heidelberg University · University Hospital Heidelberg · German Center for Lung Research

   Affiliation as printed

   Department of Diagnostic and Interventional Radiology (DIR), Heidelberg University Hospital, Heidelberg, Germany

   Translational Lung Research Center (TLRC), German Center for Lung Research (DZL), University Heidelberg, Heidelberg, Germany

10. Thomas Kuner

    Heidelberg University · German Center for Lung Research

    Affiliation as printed

    Department of Functional Neuroanatomy, Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany

    Translational Lung Research Center (TLRC), German Center for Lung Research (DZL), University Heidelberg, Heidelberg, Germany

11. Mark Oliver Wielpütz

    Heidelberg University · Universitätsmedizin Greifswald · University Hospital Heidelberg · Universität Greifswald · German Center for Lung Research

    Affiliation as printed

    Department of Diagnostic and Interventional Radiology (DIR), Heidelberg University Hospital, Heidelberg, Germany

    Diagnostic Radiology and Neuroradiology, University Medicine Greifswald, Greifswald, Germany

    Translational Lung Research Center (TLRC), German Center for Lung Research (DZL), University Heidelberg, Heidelberg, Germany

12. Marcus Alexander Mall

    Humboldt-Universität zu Berlin · German Center for Lung Research · Freie Universität Berlin · Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Pediatric Respiratory Medicine, Immunology and Critical Care Medicine, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität, Berlin, Germany

    German Center for Child and Adolescent Health (DZKJ), Partner Site Berlin, Berlin, Germany

    German Center for Lung Research (DZL), Associated Partner Site Berlin, Berlin, Germany

13. Willi Linus Wagner

    Heidelberg University · Universitätsmedizin Greifswald · University Hospital Heidelberg · Universität Greifswald · German Center for Lung Research

    Affiliation as printed

    Department of Diagnostic and Interventional Radiology (DIR), Heidelberg University Hospital, Heidelberg, Germany

    Diagnostic Radiology and Neuroradiology, University Medicine Greifswald, Greifswald, Germany

    Translational Lung Research Center (TLRC), German Center for Lung Research (DZL), University Heidelberg, Heidelberg, Germany