Primary Ciliary Dyskinesia Patient-Specific hiPSC-Derived Airway Epithelium in Air-Liquid Interface Culture Recapitulates Disease Specific Phenotypes In Vitro

Laura von Schledorn; David Puertollano Martín; Nicole Cleve; Janina Zöllner; Doris Roth; Ben Ole Staar; Jan Hegermann; Felix C Ringshausen; Janna Nawroth; Ulrich Martin; Ruth Olmer

doi:10.3390/cells12111467

Primary Ciliary Dyskinesia Patient-Specific hiPSC-Derived Airway Epithelium in Air-Liquid Interface Culture Recapitulates Disease Specific Phenotypes In Vitro

Cells. 2023 May 24;12(11):1467. doi: 10.3390/cells12111467.

Authors

Laura von Schledorn^{1

2

3}, David Puertollano Martín^{1

2

3}, Nicole Cleve^{1

2

3}, Janina Zöllner^{1

2

3}, Doris Roth⁴, Ben Ole Staar^{2

5}, Jan Hegermann^{2

6}, Felix C Ringshausen^{2

5

7}, Janna Nawroth^{4

8}, Ulrich Martin^{1

2

3}, Ruth Olmer^{1

2

3}

Affiliations

¹ Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery (HTTG), Hannover Medical School, 30625 Hannover, Germany.
² Biomedical Research in Endstage and Obstructive Lung Disease (BREATH), Member of the German Center for Lung Research (DZL), Hannover Medical School, 30625 Hannover, Germany.
³ REBIRTH-Research Center for Translational and Regenerative Medicine, Hannover Medical School, 30625 Hannover, Germany.
⁴ Helmholtz Pioneer Campus and Institute of Biological and Medical Imaging, Helmholtz Zentrum München, 85764 Neuherberg, Germany.
⁵ Department of Respiratory Medicine and Infectious Diseases, Hannover Medical School, 30625 Hannover, Germany.
⁶ Research Core Unit Electron Microscopy, Institute of Functional and Applied Anatomy, Hannover Medical School, 30625 Hannover, Germany.
⁷ European Reference Network on Rare and Complex Respiratory Diseases (ERN-LUNG), 60590 Frankfurt, Germany.
⁸ Chair of Biological Imaging at the Central Institute for Translational Cancer Research (TranslaTUM), School of Medicine, Technical University of Munich, 81675 Munich, Germany.

Abstract

Primary ciliary dyskinesia (PCD) is a rare heterogenic genetic disorder associated with perturbed biogenesis or function of motile cilia. Motile cilia dysfunction results in diminished mucociliary clearance (MCC) of pathogens in the respiratory tract and chronic airway inflammation and infections successively causing progressive lung damage. Current approaches to treat PCD are symptomatic, only, indicating an urgent need for curative therapeutic options. Here, we developed an in vitro model for PCD based on human induced pluripotent stem cell (hiPSC)-derived airway epithelium in Air-Liquid-Interface cultures. Applying transmission electron microscopy, immunofluorescence staining, ciliary beat frequency, and mucociliary transport measurements, we could demonstrate that ciliated respiratory epithelia cells derived from two PCD patient-specific hiPSC lines carrying mutations in DNAH5 and NME5, respectively, recapitulate the respective diseased phenotype on a molecular, structural and functional level.

Keywords: PCD; air-liquid interface culture; human induced pluripotent stem cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Ciliary Motility Disorders* / genetics
Epithelium
Humans
Induced Pluripotent Stem Cells*
NM23 Nucleoside Diphosphate Kinases
Phenotype
Respiratory System

Substances

NME5 protein, human
NM23 Nucleoside Diphosphate Kinases

Grants and funding

82DZL002A1/German Center for Lung Research