Human CFTR deficient iPSC-macrophages reveal impaired functional and transcriptomic response upon Pseudomonas aeruginosa infection

Claudio Rodriguez Gonzalez; Débora Basílio-Queirós; Anna-Lena Neehus; Sylvia Merkert; David Tschritter; Sinem Ünal; Jan Hegermann; Matthias Mörgelin; Jacinta Bustamante; Manuel Manfred Nietert; Ulrich Martin; Burkhard Tümmler; Antje Munder; Nico Lachmann

doi:10.3389/fimmu.2024.1397886

Human CFTR deficient iPSC-macrophages reveal impaired functional and transcriptomic response upon Pseudomonas aeruginosa infection

Front Immunol. 2024 Nov 13:15:1397886. doi: 10.3389/fimmu.2024.1397886. eCollection 2024.

Authors

Claudio Rodriguez Gonzalez¹, Débora Basílio-Queirós¹, Anna-Lena Neehus^{2

3

4

5

6}, Sylvia Merkert^{7

8

9}, David Tschritter¹⁰, Sinem Ünal^{2

3}, Jan Hegermann^{8

11}, Matthias Mörgelin¹², Jacinta Bustamante^{2

3

13

14}, Manuel Manfred Nietert¹⁰, Ulrich Martin^{7

8

9}, Burkhard Tümmler^{1

8}, Antje Munder^{1

8}, Nico Lachmann^{1

8

9

15

16}

Affiliations

¹ Department of Pediatric Pneumology, Allergology and Neonatology, Hannover Medical School, Hannover, Germany.
² Laboratory of Human Genetics of Infectious Diseases, Necker Branch, INSERM U1163, Necker Hospital for Sick Children, Paris, France.
³ Paris Cité University, Imagine Institute, Paris, France.
⁴ Division of Hematology/Oncology, Boston Children's Hospital, Harvard Medical School, Boston, MA, United States.
⁵ Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, United States.
⁶ Broad Institute of MIT and Harvard, Cambridge, MA, United States.
⁷ Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery (HTTG), Hannover Medical School, Hannover, Germany.
⁸ Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), Hannover Medical School, Hannover, Germany.
⁹ REBIRTH, Research Center for Translational and Regenerative Medicine, Hannover Medical School, Hannover, Germany.
¹⁰ Department of Medical Bioinformatics, University Medical Center Göttingen, Göttingen, Germany.
¹¹ Research Core Unit Electron Microscopy, Institute of Functional and Applied Anatomy, Hannover Medical School, Hannover, Germany.
¹² Colzyx AB, Lund, Sweden.
¹³ Study Center for Primary Immunodeficiencies, Necker Hospital for Sick Children, Assistance Publique-Hôpitaux de Paris AP-HP, Paris, France.
¹⁴ St. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, NY, United States.
¹⁵ Cluster of Excellence RESIST (EXC 2155), Hannover Medical School, Hannover, Germany.
¹⁶ Fraunhofer Institute for Toxicology and Experimental Medicine, Hannover, Germany.

Abstract

Introduction: Cystic fibrosis (CF) is a hereditary autosomal recessive disease driven by deleterious variants of the CFTR gene, leading, among other symptoms, to increased lung infection susceptibility. Mucus accumulation in the CF lung is, as of yet, considered as one important factor contributing to its colonization by opportunistic pathogens such as Pseudomonas aeruginosa. However, in recent years evidence was provided that alveolar macrophages, which form the first line of defense against airborne pathogens, seem to be intrinsically defective with regard to bactericidal functionality in the CF lung. To assess the impact of CFTR deficiency in human macrophages only insufficient systems are available.

Methods: To address this problem and to evaluate the role of CFTR in human macrophages, we successfully differentiated human induced pluripotent stem cells (iPSC) from a CF p.Phe508del homozygous individual and a healthy donor into primitive macrophages (iMac^ΔF508 and iMac^WT), respectively, and compared the bactericidal functionality in the relevant cell type.

Results: iMac^ΔF508 showed impaired P. aeruginosa clearance and intracellular killing capacity in comparison to iMac^WT. Furthermore, iMac^ΔF508 exhibited a less acidic lysosomal pH, and upon P. aeruginosa infection, there were signs of mitochondrial fragmentation and autophagosome formation together with a hyperinflammatory phenotype and deficient type I interferon response.

Conclusion: In summary, we present a defective phenotype in iMac^ΔF508 upon P. aeruginosa infection, which will constitute an ideal platform to further study the role of macrophages in the context of CF.

Keywords: Pseudomonas aeruginosa; cystic fibrosis; iPSC; infection; lung immunity; macrophages.

MeSH terms

Cell Differentiation / genetics
Cells, Cultured
Cystic Fibrosis Transmembrane Conductance Regulator* / deficiency
Cystic Fibrosis Transmembrane Conductance Regulator* / genetics
Cystic Fibrosis* / genetics
Cystic Fibrosis* / immunology
Cystic Fibrosis* / microbiology
Humans
Induced Pluripotent Stem Cells* / metabolism
Macrophages / immunology
Macrophages / metabolism
Macrophages / microbiology
Pseudomonas Infections* / genetics
Pseudomonas Infections* / immunology
Pseudomonas aeruginosa* / immunology
Pseudomonas aeruginosa* / physiology
Transcriptome*

Substances

Cystic Fibrosis Transmembrane Conductance Regulator
CFTR protein, human

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study was made possible by a non-clinical research grant from the Mukoviszidose e.V. to AM and NL (project number 1905). Additional funding was provided by the German Center of Lung Research (DZL) to AM. This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (grant agreement No. 852178) (grant agreement No. 101100859 “iPYRO” and No. 101158172 “iMAClung). The work is also funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy -EXC 2155 -project number 390874280 and REBIRTH Research Center for Translational Regenerative Medicine “Förderung aus Mitteln des Niedersächsischen Vorab” (grant: ZN3340). The work is also funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) LA 3680/9-1. A-LN was supported by the international PhD program of the Imagine Institute, the Bettencourt-Schueller Foundation, the fin de thèse program of the FRM (FDT202204015102) and the EMBO Postdoctoral Fellowship (ALTF 209-2024). MN was supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) project number 315063128.