Gain-of-function variants in the ion channel gene TRPM3 underlie a spectrum of neurodevelopmental disorders

  1. Lydie Burglen
  2. Evelien Van Hoeymissen
  3. Leila Qebibo
  4. Magalie Barth
  5. Newell Belnap
  6. Felix Boschann
  7. Christel Depienne
  8. Katrien De Clercq
  9. Andrew GL Douglas
  10. Mark P Fitzgerald
  11. Nicola Foulds
  12. Catherine Garel
  13. Ingo Helbig
  14. Katharina Held
  15. Denise Horn
  16. Annelies Janssen
  17. Angela M Kaindl
  18. Vinodh Narayanan
  19. Christine Prager
  20. Mailys Rupin
  21. Alexandra Afenjar
  22. Siyuan Zhao
  23. Vincent Th Ramaekers
  24. Sarah M Ruggiero
  25. Simon Thomas
  26. Stéphanie Valence
  27. Lionel Van Maldergem
  28. Tibor Rohacs
  29. Diana Rodriguez
  30. David Dyment
  31. Thomas Voets  Is a corresponding author
  32. Joris Vriens  Is a corresponding author
  1. INSERM UMR 1163, France
  2. KU Leuven, Belgium
  3. Hôpitaux Universitaires Paris-Ouest, France
  4. Centre Hospitalier Universitaire d'Angers, France
  5. Translational Genomics Research Institute, United States
  6. Charité - Universitäts medizin Berlin, Germany
  7. Essen University Hospital, United States
  8. University Hospital Southampton NHS Foundation Trust, United Kingdom
  9. Children's Hospital of Philadelphia, United States
  10. Charité - Universitätsmedizin Berlin, Germany
  11. Rutgers, The State University of New Jersey, United States
  12. University of Liège, Belgium
  13. Salisbury District Hospital, United Kingdom
  14. Centre Hospitalier Universitaire de Besançon, France
  15. University of Ottawa, Canada
  16. VIB-KU Leuven Center for Brain & Disease Research, Belgium

Abstract

TRPM3 is a temperature- and neurosteroid-sensitive plasma membrane cation channel expressed in a variety of neuronal and non-neuronal cells. Recently, rare de novo variants in TRPM3 were identified in individuals with developmental and epileptic encephalopathy (DEE), but the link between TRPM3 activity and neuronal disease remains poorly understood. We previously reported that two disease-associated variants in TRPM3 lead to a gain of channel function (Van Hoeymissen et al., 2020; Zhao et al., 2020). Here, we report a further ten patients carrying one of seven additional heterozygous TRPM3 missense variants. These patients present with a broad spectrum of neurodevelopmental symptoms, including global developmental delay, intellectual disability, epilepsy, musculo-skeletal anomalies, and altered pain perception. We describe a cerebellar phenotype with ataxia or severe hypotonia, nystagmus, and cerebellar atrophy in more than half of the patients. All disease-associated variants exhibited a robust gain-of-function phenotype, characterized by increased basal activity leading to cellular calcium overload and by enhanced responses to the neurosteroid ligand pregnenolone sulphate, when co-expressed with wild-type TRPM3 in mammalian cells. The antiseizure medication primidone, a known TRPM3 antagonist, reduced the increased basal activity of all mutant channels. These findings establish gain-of-function of TRPM3 as the cause of a spectrum of autosomal dominant neurodevelopmental disorders with frequent cerebellar involvement in humans, and provide support for the evaluation of TRPM3 antagonists as a potential therapy.

Data availability

Raw data for the following figures are made available via figshare (https://doi.org/10.6084/m9.figshare.21799604): Figure 1 - Figure Supplement 1; Figure 3; Figure 3 - Figure Supplement 1, 2, 3 and 4; Figure 4; Figure 4 - Figure Supplement 1, 2 and 3.

The following data sets were generated

Article and author information

Author details

  1. Lydie Burglen

    Developmental Brain Disorders Laboratory, INSERM UMR 1163, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  2. Evelien Van Hoeymissen

    Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3897-8998
  3. Leila Qebibo

    Département de Génétique, Hôpitaux Universitaires Paris-Ouest, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  4. Magalie Barth

    Department of Genetics, Centre Hospitalier Universitaire d'Angers, Angers, France
    Competing interests
    The authors declare that no competing interests exist.
  5. Newell Belnap

    Neurogenomics Division, Translational Genomics Research Institute, Phoenix, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Felix Boschann

    Institute of Medical Genetics and Human Genetics, Charité - Universitäts medizin Berlin, Berlin, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Christel Depienne

    Institute of Human Genetics, Essen University Hospital, Essen, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Katrien De Clercq

    Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium
    Competing interests
    The authors declare that no competing interests exist.
  9. Andrew GL Douglas

    University Hospital Southampton NHS Foundation Trust, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  10. Mark P Fitzgerald

    Children's Hospital of Philadelphia, Philadelphia, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Nicola Foulds

    Wessex Clinical Genetics Service, University Hospital Southampton NHS Foundation Trust, Wessex, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  12. Catherine Garel

    Département de Génétique, Hôpitaux Universitaires Paris-Ouest, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  13. Ingo Helbig

    Children's Hospital of Philadelphia, Philadelphia, United States
    Competing interests
    The authors declare that no competing interests exist.
  14. Katharina Held

    Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium
    Competing interests
    The authors declare that no competing interests exist.
  15. Denise Horn

    Institute of Medical Genetics and Human Genetics, Charité - Universitäts medizin Berlin, Berlin, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0870-8911
  16. Annelies Janssen

    Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6735-8248
  17. Angela M Kaindl

    Institute of Cell Biology and Neurobiology, Charité - Universitätsmedizin Berlin, Berlin, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9454-206X
  18. Vinodh Narayanan

    Neurogenomics Division, Translational Genomics Research Institute, Phoenix, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0658-3847
  19. Christine Prager

    Institute of Medical Genetics and Human Genetics, Charité - Universitäts medizin Berlin, Berlin, Germany
    Competing interests
    The authors declare that no competing interests exist.
  20. Mailys Rupin

    Department of Neuropediatrics, Centre Hospitalier Universitaire d'Angers, Angers, France
    Competing interests
    The authors declare that no competing interests exist.
  21. Alexandra Afenjar

    Developmental Brain Disorders Laboratory, INSERM UMR 1163, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  22. Siyuan Zhao

    Department of Pharmacology, Physiology and Neuroscience, Rutgers, The State University of New Jersey, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2005-9440
  23. Vincent Th Ramaekers

    Division Neuropediatrics, University of Liège, Liège, Belgium
    Competing interests
    The authors declare that no competing interests exist.
  24. Sarah M Ruggiero

    Children's Hospital of Philadelphia, Philadelphia, United States
    Competing interests
    The authors declare that no competing interests exist.
  25. Simon Thomas

    Wessex Regional Genetics Laboratory, Salisbury District Hospital, Wessex, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  26. Stéphanie Valence

    Département de Génétique, Hôpitaux Universitaires Paris-Ouest, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  27. Lionel Van Maldergem

    Centre de Génétique Humaine, Centre Hospitalier Universitaire de Besançon, Besancon, France
    Competing interests
    The authors declare that no competing interests exist.
  28. Tibor Rohacs

    Department of Pharmacology, Physiology and Neuroscience, Rutgers, The State University of New Jersey, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3580-2575
  29. Diana Rodriguez

    Département de Génétique, Hôpitaux Universitaires Paris-Ouest, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  30. David Dyment

    Children's Hospital of Eastern Ontario Research Institute, University of Ottawa, Ottawa, Canada
    Competing interests
    The authors declare that no competing interests exist.
  31. Thomas Voets

    VIB-KU Leuven Center for Brain & Disease Research, Leuven, Belgium
    For correspondence
    thomas.voets@kuleuven.be
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5526-5821
  32. Joris Vriens

    Department of Development and Regeneration, KU Leuven, Leuven, Belgium
    For correspondence
    Joris.Vriens@kuleuven.be
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2502-0409

Funding

Flanders' FOOD (G.0D1417N)

  • Joris Vriens

Flanders' FOOD (G.084515N)

  • Joris Vriens

Flanders' FOOD (G.0A6719N)

  • Joris Vriens

Flanders' FOOD (11E782)

  • Evelien Van Hoeymissen

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Human subjects: The study was performed in accordance with the guidelines specified by the institutional review boards and ethics committees at each institution. Information of institutional protocols are provided in the section of Material & Methods. All parents agreed on sharing and publicing the patients' information.Patients information:patient 1, 3, 4, 7: Written informed consent was obtained from the parents of the probands for molecular genetic analysis and possible publication of the anonymized clinical data. The study was done in accordance with local research and ethics requirements.patient 2: Parents signed an informed consent, received a genetic counselling before and after the analysis, and the genetic study was performed in accordance with German and French ethical requirements and laws.patient 5: UK ethical approval by the Cambridge South Research Ethics Committee (10/H0305/83)patient 6: outine clinical care within the UK National Health Service, and so no specific institutional ethical approval was requiredpatient 8: Declaration of Helsinki with local approval by the Children's Hospital of Philadelphia (CHOP) Institutional Review Board (IRB 15-12226).patient 9: The participating family signed the IRB research protocol of the University of Pennsylvania division of Neurologypatient 10: The study protocol and consent documents were approved by the Western Institutional Review Board (WIRB # 20120789). The retrospective analysis of epilepsy patient data was approved by the local ethics committees of the Charité (approval no. EA2/084/18)

Reviewing Editor

  1. Andres Jara-Oseguera, The University of Texas at Austin, United States

Publication history

  1. Received: June 13, 2022
  2. Accepted: December 7, 2022
  3. Accepted Manuscript published: January 17, 2023 (version 1)
  4. Accepted Manuscript updated: January 19, 2023 (version 2)

Copyright

© 2023, Burglen et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 533
    Page views
  • 198
    Downloads
  • 0
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Lydie Burglen
  2. Evelien Van Hoeymissen
  3. Leila Qebibo
  4. Magalie Barth
  5. Newell Belnap
  6. Felix Boschann
  7. Christel Depienne
  8. Katrien De Clercq
  9. Andrew GL Douglas
  10. Mark P Fitzgerald
  11. Nicola Foulds
  12. Catherine Garel
  13. Ingo Helbig
  14. Katharina Held
  15. Denise Horn
  16. Annelies Janssen
  17. Angela M Kaindl
  18. Vinodh Narayanan
  19. Christine Prager
  20. Mailys Rupin
  21. Alexandra Afenjar
  22. Siyuan Zhao
  23. Vincent Th Ramaekers
  24. Sarah M Ruggiero
  25. Simon Thomas
  26. Stéphanie Valence
  27. Lionel Van Maldergem
  28. Tibor Rohacs
  29. Diana Rodriguez
  30. David Dyment
  31. Thomas Voets
  32. Joris Vriens
(2023)
Gain-of-function variants in the ion channel gene TRPM3 underlie a spectrum of neurodevelopmental disorders
eLife 12:e81032.
https://doi.org/10.7554/eLife.81032

Further reading

    1. Cell Biology
    2. Chromosomes and Gene Expression
    Liangyu Zhang, Weston T Stauffer ... Abby F Dernburg
    Research Article

    Meiotic chromosome segregation relies on synapsis and crossover recombination between homologous chromosomes. These processes require multiple steps that are coordinated by the meiotic cell cycle and monitored by surveillance mechanisms. In diverse species, failures in chromosome synapsis can trigger a cell cycle delay and/or lead to apoptosis. How this key step in 'homolog engagement' is sensed and transduced by meiotic cells is unknown. Here we report that in C. elegans, recruitment of the Polo-like kinase PLK-2 to the synaptonemal complex triggers phosphorylation and inactivation of CHK-2, an early meiotic kinase required for pairing, synapsis, and double-strand break induction. Inactivation of CHK-2 terminates double-strand break formation and enables crossover designation and cell cycle progression. These findings illuminate how meiotic cells ensure crossover formation and accurate chromosome segregation.

    1. Cell Biology
    2. Physics of Living Systems
    Christa Ringers, Stephan Bialonski ... Nathalie Jurisch-Yaksi
    Research Article

    Motile cilia are hair-like cell extensions that beat periodically to generate fluid flow along various epithelial tissues within the body. In dense multiciliated carpets, cilia were shown to exhibit a remarkable coordination of their beat in the form of traveling metachronal waves, a phenomenon which supposedly enhances fluid transport. Yet, how cilia coordinate their regular beat in multiciliated epithelia to move fluids remains insufficiently understood, particularly due to lack of rigorous quantification. We combine experiments, novel analysis tools, and theory to address this knowledge gap. To investigate collective dynamics of cilia, we studied zebrafish multiciliated epithelia in the nose and the brain. We focused mainly on the zebrafish nose, due to its conserved properties with other ciliated tissues and its superior accessibility for non-invasive imaging. We revealed that cilia are synchronized only locally and that the size of local synchronization domains increases with the viscosity of the surrounding medium. Even though synchronization is local only, we observed global patterns of traveling metachronal waves across the zebrafish multiciliated epithelium. Intriguingly, these global wave direction patterns are conserved across individual fish, but different for left and right nose, unveiling a chiral asymmetry of metachronal coordination. To understand the implications of synchronization for fluid pumping, we used a computational model of a regular array of cilia. We found that local metachronal synchronization prevents steric collisions, cilia colliding with each other, and improves fluid pumping in dense cilia carpets, but hardly affects the direction of fluid flow. In conclusion, we show that local synchronization together with tissue-scale cilia alignment coincide and generate metachronal wave patterns in multiciliated epithelia, which enhance their physiological function of fluid pumping.