Abstract

Primary ciliary dyskinesia (PCD) is an inherited disorder characterized by recurrent infections of the upper and lower respiratory tract, reduced fertility in males and situs inversus in about 50% of affected individuals (Kartagener syndrome). It is caused by motility defects in the respiratory cilia that are responsible for airway clearance, the flagella that propel sperm cells and the nodal monocilia that determine left-right asymmetry1. Recessive mutations that cause PCD have been identified in genes encoding components of the outer dynein arms, radial spokes and cytoplasmic pre-assembly factors of axonemal dyneins, but these mutations account for only about 50% of cases of PCD. We exploited the unique properties of dog populations to positionally clone a new PCD gene, CCDC39. We found that loss-of-function mutations in the human ortholog underlie a substantial fraction of PCD cases with axonemal disorganization and abnormal ciliary beating. Functional analyses indicated that CCDC39 localizes to ciliary axonemes and is essential for assembly of inner dynein arms and the dynein regulatory complex.

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Acknowledgements

This work was supported by grants from the European Union (LUPA IP) and from the Police Scientifique Fédérale de Belgique (GENFUNC PAI) (to M.G.), from the Legs Poix from the Chancellerie des Universités, the Assistance Publique-Hôpitaux de Paris (PHRC AOM06053, P060245) and the Agence Nationale pour la Recherche (ANR-05-MRAR-022-01) (to S.A.), the US National Institutes of Health (HD04260, DK072301 and DK075972 (to N.K.) and DK079541 (to E.E.D.)) and by grants from the “Deutsche Forschungsgemeinschaft” DFG Om 6/4, GRK1104, SFB592, and the European Community (EU-CILIA; SYS-CILIA) (to H.O.). A.C.M. is a fellow from the FRIA. N.K. is the Jean and George W. Brumley Professor. Y.M. benefits from a postdoctoral fellowship to study abroad from the Japanese Society for the Promotion of Science (JSPS). We thank the Bobtail breeders for assistance; patients and their family members whose cooperation made this study possible; all referring physicians; the German patient support group “Kartagener Syndrom und Primaere Ciliaere Dyskinesie e.V.”; K. Nakamura and the GIGA-R genomics platform for their contribution to sequencing; E. Ostrander for samples from healthy Old English Sheepdogs; the Unité de Recherche Clinique (URC) Est (AP-HP, Hôpital Saint-Antoine, Paris, France) for support; and A. Heer, C. Reinhard, C. Kopp, K. Sutter, M. Petry, C. Tessmer, A.-M. Vojtek and S. Franz for technical assistance.

Author information

Author notes

    • Anne-Christine Merveille
    • , Erica E Davis
    • , Anita Becker-Heck
    •  & Marie Legendre

    These authors contributed equally to this work.

    • Anne-Sophie Lequarré
    • , Nicholas Katsanis
    • , Heymut Omran
    •  & Serge Amselem

    These authors jointly supervised this work.

Affiliations

  1. Unit of Animal Genomics, Groupe Interdisciplinaire de Genomique Appliquee-Recherche (GIGA-R) and Faculty of Veterinary Medicine, University of Liège (B34), Liège, Belgium.

    • Anne-Christine Merveille
    • , Géraldine Bataille
    • , Yukihide Momozawa
    • , Michel Georges
    •  & Anne-Sophie Lequarré
  2. Center for Human Disease Modeling, Department of Cell Biology, Duke University Medical Center, Durham, North Carolina, USA.

    • Erica E Davis
    •  & Nicholas Katsanis
  3. Department of Pediatrics and Adolescent Medicine, University Hospital Freiburg, Freiburg, Germany.

    • Anita Becker-Heck
    • , Manfred Fliegauf
    • , Judith Horvath
    • , Niki Tomas Loges
    • , Heike Olbrich
    • , Miriam Schmidts
    •  & Heymut Omran
  4. Faculty of Biology, Albert-Ludwigs-University, Freiburg, Germany.

    • Anita Becker-Heck
  5. Department of General Pediatrics, University Children's Hospital Münster, Münster, Germany.

    • Anita Becker-Heck
    • , Niki Tomas Loges
    •  & Heymut Omran
  6. Institut National de la Santé et de la Recherche Médicale (INSERM) U.933, Université Pierre et Marie Curie-Paris 6 and Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Armand-Trousseau, Paris, France.

    • Marie Legendre
    • , Philippe Duquesnoy
    • , Denise Escalier
    • , Estelle Escudier
    • , Guy Montantin
    • , Heike Olbrich
    • , Jean-François Papon
    • , Henrique Tenreiro
    •  & Serge Amselem
  7. Department of Pediatrics, Ziv Medical Center, Safed, Israel.

    • Israel Amirav
  8. Rapaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.

    • Israel Amirav
  9. Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas, USA.

    • John Belmont
    •  & Jeffrey A Towbin
  10. AP-HP, Hôpital Armand-Trousseau, Service d′explorations fonctionnelles respiratoires, Paris, France.

    • Nicole Beydon
  11. Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Liège, Belgium.

    • Frédéric Billen
    • , Cécile Clercx
    •  & Stephane Deleuze
  12. AP-HP, Hôpital Armand-Trousseau, Unité de pneumologie pédiatrique, Centre de Référence des Maladies Respiratoires Rares, Paris, France.

    • Annick Clément
  13. AP-HP, Hôpital Intercommunal et Groupe Hospitalier Henri Mondor-Albert Chenevier, Service d'ORL et de chirurgie cervico-faciale, Créteil, France.

    • André Coste
    •  & Jean-François Papon
  14. Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.

    • Rachelle Crosbie
    •  & Kent Hill
  15. AP-HP, Groupe Hospitalier Necker-Enfants Malades, Service de pneumologie et d'allergologie pédiatriques, Paris, France.

    • Jacques de Blic
  16. Department of Otorhinolaryngology, Head and Neck Surgery, University Hospitals Leuven, Leuven, Belgium.

    • Mark Jorissen
  17. AP-HP, Hôpital Armand-Trousseau, Centre d'investigation de l'asthme et des allergies, Paris, France.

    • Jocelyne Just
  18. Institut für Molekularbiologie, Medizinische Hochschule Hannover, Hannover, Germany.

    • Andreas Kispert
  19. Centre National de Génotypage, Evry, France.

    • Mark Lathrop
    •  & Diana Zelenika
  20. Hôpital Nord, Service de réanimation pédiatrique, Centre Hospitalier Universitaire de Saint-Etienne, Saint-Etienne, France.

    • June K Marthin
    •  & Isabelle Rayet
  21. Copenhagen University Hospital, Rigshospitalet, Danish Primary Ciliary Dyskinesia Center, Pediatric Pulmonary Service, Copenhagen, Denmark.

    • Kim G Nielsen
  22. AP-HP, Hôpital Armand-Trousseau, Service d'ORL et de chirurgie cervico-faciale pédiatrique, Paris, France.

    • Gilles Roger
  23. Department of Tumor Virology, German Cancer Research Center, Heidelberg, Germany.

    • Hanswalter Zentgraf

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Contributions

The positional cloning of CCDC39 in the dog was performed by A.-C.M. and G.B. Genome-wide SNP genotyping was conducted at CNG under supervision of M. Lathrop and D.Z. Mining the ciliome databases was conducted by E.E.D. Experiments in the zebrafish were conducted by E.E.D. In situ hybridization in the mouse was conducted by A.K. and H.O. qRT-PCR on human samples was conducted by M. Legendre, P.D., G.M. and H.T. Sequencing of CCDC39 in human subjects, including high-throughput sequencing, was conducted by A.-C.M., G.B., Y.M., A.B.-H., M. Legendre, E.E., P.D., G.M. and H.T. Identifying the p.Glu390SerfsX6 mutation was realized by M. Legendre, P.D., G.M. and H.T. Haplotype analysis to determine founder status of CCDC39 mutations was conducted by M. Legendre and P.D. TEM analysis was conducted by M.J. (dog), E.E. and D.E. (French cohort), and by K.G.N., J.K.M., H.O. and routine laboratories (German cohort). High-resolution immunofluorescence analyses were done by A.B.-H., M.F., J.H. and N.T.L. Immunoblotting analyses were done by A.B.-H. High-speed video analyses were conducted by H.O., N.T.L. and A.B.-H. Monoclonal anti-GAS11 antibody was produced by A.B.-H. and H.Z. Polyclonal anti-GAS11 antibodies were provided by K.H. and R.C. Clinical examination and collection of the canine PCD cases was conducted by F.B., C.C. and S.D. M.G., A.-S.L., N.K., H.O. and S.A. designed experiments, analyzed data and wrote the manuscript. All remaining authors as well as H.O., K.G.N. and J.K.M. examined and contributed samples from individuals with PCD or heterotaxia.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Michel Georges.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Note, Supplementary Figures 1–9 and Supplementary Table 1

Videos

  1. 1.

    Supplemental Video 1

    Ciliary beating pattern of PCD patient OP122 with CCDC39 mutations assessed by high-speed videomicroscopy analyses of respiratory cells obtained by nasal brushing biopsy

  2. 2.

    Supplemental Video 2

    Ciliary beating pattern of a healthy control individual assessed by high-speed videomicroscopy analyses of respiratory cells obtained by nasal brushing biopsy

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DOI

https://doi.org/10.1038/ng.726

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