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CCDC103 mutations cause primary ciliary dyskinesia by disrupting assembly of ciliary dynein arms

Nature Genetics volume 44, pages 714–719 (2012)Cite this article

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Abstract

Cilia are essential for fertilization, respiratory clearance, cerebrospinal fluid circulation and establishing laterality1. Cilia motility defects cause primary ciliary dyskinesia (PCD, MIM244400), a disorder affecting 1:15,000–30,000 births. Cilia motility requires the assembly of multisubunit dynein arms that drive ciliary bending2. Despite progress in understanding the genetic basis of PCD, mutations remain to be identified for several PCD-linked loci3. Here we show that the zebrafish cilia paralysis mutant schmalhans (smhtn222) encodes the coiled-coil domain containing 103 protein (Ccdc103), a foxj1a-regulated gene product. Screening 146 unrelated PCD families identified individuals in six families with reduced outer dynein arms who carried mutations in CCDC103. Dynein arm assembly in smh mutant zebrafish was rescued by wild-type but not mutant human CCDC103. Chlamydomonas Ccdc103/Pr46b functions as a tightly bound, axoneme-associated protein. These results identify Ccdc103 as a dynein arm attachment factor that causes primary ciliary dyskinesia when mutated.

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Figure 1: The smh mutation causes cilia paralysis and absent dynein arms.
Figure 2: Pedigrees of families carrying CCDC103 loss-of-function mutations.
Figure 3: Functional assay of human CCDC103 mutant alleles in smh embryos.
Figure 4: Localization of DNAH5, DNAI2 and DNAH9 in respiratory epithelial cells from a patient with PCD carrying the CCDC103 loss-of-function mutation.
Figure 5: Ccdc103 homodimers assemble with dynein light chain 2 in the cytoplasm and bind tightly to cilia axonemes.

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Article 06 February 2020

Stylianos E. Antonarakis, Brian G. Skotko, … Roger H. Reeves

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Acknowledgements

We thank the patients and their families for their participation and acknowledge support from the PCD Family Support Group (UK) and the German patient support group 'Kartagener Syndrom und Primaere Ciliaere Dyskinesie e.V.' . We thank R. Mark Gardiner and E. Moya for their help and involvement in the study. We thank R. Hirst and A. Rutman for electron microscopy. We thank A. Heer, C. Warmt, D. Nergenau, R.S. Patel-King and L. Luo for excellent technical assistance. This work was funded by US National Institutes of Health grants DK053093 (I.A.D.) and GM051293 (S.M.K.), National Research Service Award fellowship grant F32DK083868 (J.R.P.), an American Heart Association Established Investigator Award (A.F.S.), Deutsche Forschungsgemeinschaft (DFG) grants (DFG Om 6/4, GRK1109 and SFB592), EU grant SYSCILIA, IZKF-project (Om2/009/12), and Kindness for Kids grant (H. Omran), the Medical Research Council UK, the Wellcome Trust and the Fondation Milena Carvajal–ProKartagener.

Author information

Author notes

  1. Jennifer R Panizzi and Anita Becker-Heck: These authors contributed equally to this work.

  2. Stephen M King, Heymut Omran and Iain A Drummond: These authors jointly directed this work.

Authors and Affiliations

  1. Nephrology Division, Massachusetts General Hospital, Charlestown, Massachusetts, USA

    Jennifer R Panizzi, Yan Liu, Narendra Pathak, Nathan Hellman, Amar Gupta & Iain A Drummond

  2. Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA

    Jennifer R Panizzi, Narendra Pathak, Nathan Hellman & Iain A Drummond

  3. Department of Pediatrics, University Hospital Freiburg, Freiburg, Germany

    Anita Becker-Heck & Karsten Häffner

  4. Klinik und Poliklinik fuer Kinder und Jugendmedizin Allgemeine Paediatrie, Universitätsklinikum Münster, Münster, Germany

    Anita Becker-Heck, Niki T Loges, Heike Olbrich, Claudius Werner & Heymut Omran

  5. Molecular Medicine Unit, University College London, Institute of Child Health, London, UK

    Victoria H Castleman, Miriam Schmidts & Hannah M Mitchison

  6. Leeds Institute of Molecular Medicine, Wellcome Trust Brenner Building, St James's University Hospital, Leeds, UK

    Dalal A Al-Mutairi & Eamonn Sheridan

  7. Department of Pathology, Kuwait University, Faculty of Medicine, Kuwait University, Kuwait

    Dalal A Al-Mutairi

  8. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA

    Christina Austin-Tse & Iain A Drummond

  9. General and Adolescent Paediatrics Unit, University College London, Institute of Child Health, London, UK

    Rahul Chodhari & Eddie M K Chung

  10. Renal Division, University Hospital Freiburg, Freiburg, Germany

    Albrecht Kramer-Zucker

  11. Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, New York, USA

    Felix Olale

  12. Department of Molecular Biology, Princeton University, Princeton, New Jersey, USA

    Rebecca D Burdine

  13. Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA

    Alexander F Schier

  14. Department of Infection, Immunity and Inflammation, University of Leicester, Leicester, UK

    Christopher O'Callaghan

  15. Genome Centre Cologne at Max Planck Institute for Plant Breeding Research, Köln, Germany

    Richard Reinhardt

  16. Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, Connecticut, USA

    Stephen M King

Authors
  1. Jennifer R Panizzi
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Contributions

J.R.P., Y.L., C.A.-T., A.G., N.H., A.K.-Z., R.D.B., F.O., A.F.S., N.P. and I.A.D. performed genetic mapping and Ccdc103 characterization in zebrafish. A.B.-H., V.H.C., D.A.A.-M., N.T.L., E.S., M.S., H. Olbrich, C.W., K.H., R.C., C.O., E.M.K.C., R.R., H.M.M. and H. Omran conducted studies with human patient samples. S.M.K. performed Chlamydomonas studies. J.R.P., A.B.-H., S.M.K., H. Omran and I.A.D. prepared the manuscript.

Corresponding authors

Correspondence to Stephen M King, Heymut Omran or Iain A Drummond.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Tables 1–7 and Supplementary Figures 1–16. (PDF 1406 kb)

Supplementary Video 1

Pronephric cilia motility in wildtype 2.5 dpf proximal tubules. Movies were acquired at 250 frames per second (fps) and slowed to 15 fps for viewing. Anterior is to the left. (MOV 3690 kb)

Supplementary Video 2

Pronephric cilia paralysis in schmalhans 2.5 dpf mutant proximal tubules. Movies were acquired at 250 frames per second (fps) and slowed to 15 fps for viewing. Anterior is to the left. (MOV 1799 kb)

Supplementary Video 3

Olfactory placode cilia motility in wildtype 3 dpf embryos. Movies were acquired at 250 frames per second (fps) and slowed to 15 fps for viewing. (MOV 786 kb)

Supplementary Video 4

Olfactory placode cilia paralysis in schmalhans mutant 3 dpf embryos. Movies were acquired at 250 frames per second (fps) and slowed to 15 fps for viewing. (MOV 1461 kb)

Supplementary Movie 5

Spinal canal cilia motility in wildtype 3 dpf embryos. Movies were acquired at 250 frames per second (fps) and slowed to 15 fps for viewing. Anterior is to the left. (MOV 3088 kb)

Supplementary Video 6

Spinal canal cilia paralysis in schmalhans mutant 3 dpf embryos. Movies were acquired at 250 frames per second (fps) and slowed to 15 fps for viewing. Anterior is to the left. (MOV 2619 kb)

Supplementary Video 7

Restoration of olfactory placode cilia motility in genotypically mutant schmalhans embryo by ccdc103 mRNA injection. Movies were acquired at 240 frames per second (fps) and slowed to 15 fps for viewing. (MOV 1971 kb)

Supplementary Video 8

Pronephric cilia paralysis in ccdc103 morpholino knockdown 2.5 dpf proximal tubules. Movies were acquired at 250 frames per second (fps) and slowed to 15 fps for viewing. Anterior is to the upper right corner. (MOV 2272 kb)

Supplementary Video 9

Normal human ciliated nasal epithelium show robust cilia motility and wave form. Video was acquired at 256 frames per second and the ciliary beat pattern was evaluated by slow motion playback at 30 fps. (MOV 532 kb)

Supplementary Video 10

Human ciliated nasal epithelium cells from patient OP-1193 II1 carrying the homozygous p.Gly128fs25* mutation. Cilia are completely paralysed. Video was acquired at 125 frames per second and the ciliary beat pattern was evaluated by slow motion playback at 30 fps. (MOV 760 kb)

Supplementary Video 11

Human ciliated nasal epithelium cells from patient OP-32II1 carrying the p.His154Pro CCDC103 missense mutation. Cilia show reduced beat amplitude. Video was acquired at 125 frames per second and the ciliary beat pattern was evaluated by slow motion playback at 30 fps. (MOV 290 kb)

Supplementary Video 12

Human ciliated nasal epithelium cells from patient OP-32II2 carrying the p.His154Pro CCDC103 missense mutation. Cilia are paralysed or show reduced beat frequency, reduced beat amplitude and lack of beat coordination. Video was acquired at 125 frames per second and the ciliary beat pattern was evaluated by slow motion playback at 30 fps. (MOV 653 kb)

Supplementary Video 13

Normal human ciliated nasal epithelium cells show robust cilia motility and wave form. Video was acquired at 125 frames per second and the ciliary beat pattern was evaluated by slow motion playback at 30 fps. (MOV 183 kb)

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Panizzi, J., Becker-Heck, A., Castleman, V. et al. CCDC103 mutations cause primary ciliary dyskinesia by disrupting assembly of ciliary dynein arms. Nat Genet 44, 714–719 (2012). https://doi.org/10.1038/ng.2277

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