Seizures, behavioral deficits and adverse drug responses in two new genetic mouse models of HCN1 epileptic encephalopathy

  1. Andrea Merseburg
  2. Jacquelin Kasemir
  3. Eric W Buss
  4. Felix Leroy
  5. Tobias Bock
  6. Alessandro Porro
  7. Anastasia Barnett
  8. Simon E Tröder
  9. Birgit Engeland
  10. Malte Stockebrand
  11. Anna Moroni
  12. Steve Siegelbaum
  13. Dirk Isbrandt
  14. Bina Santoro  Is a corresponding author
  1. German Center for Neurodegenerative Diseases, Germany
  2. Columbia University, United States
  3. University of Milan, Italy
  4. University of Cologne, Germany

Abstract

De novo mutations in voltage- and ligand-gated channels have been associated with an increasing number of cases of developmental and epileptic encephalopathies, which often fail to respond to classic antiseizure medications. Here, we examine two knock-in mouse models replicating de novo sequence variations in the HCN1 voltage-gated channel gene, p.G391D and p.M153I (Hcn1G380D/+ and Hcn1M142I/+ in mouse), associated with severe drug-resistant neonatal- and childhood-onset epilepsy, respectively. Heterozygous mice from both lines displayed spontaneous generalized tonic-clonic seizures. Animals replicating the p.G391D variant had an overall more severe phenotype, with pronounced alterations in the levels and distribution of HCN1 protein, including disrupted targeting to the axon terminals of basket cell interneurons. In line with clinical reports from patients with pathogenic HCN1 sequence variations, administration of the antiepileptic Na+ channel antagonists lamotrigine and phenytoin resulted in the paradoxical induction of seizures in both mouse lines, consistent with an effect to further impair inhibitory neuron function. We also show that these variants can render HCN1 channels unresponsive to classic antagonists, indicating the need to screen mutated channels to identify novel compounds with diverse mechanism of action. Our results underscore the necessity of tailoring effective therapies for specific channel gene variants, and how strongly validated animal models may provide an invaluable tool towards reaching this objective.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for Figures 2, 3, 5, 9 and 10. Microarray expression data have been deposited in GEO under accession code GSE209630.

The following data sets were generated

Article and author information

Author details

  1. Andrea Merseburg

    Experimental Neurophysiology, German Center for Neurodegenerative Diseases, Bonn, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0630-6564
  2. Jacquelin Kasemir

    Experimental Neurophysiology, German Center for Neurodegenerative Diseases, Bonn, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9176-5241
  3. Eric W Buss

    Department of Neuroscience, Columbia University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0473-4717
  4. Felix Leroy

    Department of Neuroscience, Columbia University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Tobias Bock

    Department of Neuroscience, Columbia University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Alessandro Porro

    Department of Biosciences, University of Milan, Milan, Italy
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4845-6165
  7. Anastasia Barnett

    Department of Neuroscience, Columbia University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Simon E Tröder

    Faculty of Medicine, University of Cologne, Cologne, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. Birgit Engeland

    Experimental Neurophysiology, German Center for Neurodegenerative Diseases, Bonn, Germany
    Competing interests
    The authors declare that no competing interests exist.
  10. Malte Stockebrand

    Experimental Neurophysiology, German Center for Neurodegenerative Diseases, Bonn, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9009-137X
  11. Anna Moroni

    Department of Biosciences, University of Milan, Milan, Italy
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1860-406X
  12. Steve Siegelbaum

    Department of Neuroscience, Columbia University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. Dirk Isbrandt

    Experimental Neurophysiology, German Center for Neurodegenerative Diseases, Cologne, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4720-1016
  14. Bina Santoro

    Department of Neuroscience, Columbia University, New York, United States
    For correspondence
    bs73@columbia.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4277-1992

Funding

National Institutes of Health (NS106983)

  • Steve Siegelbaum

National Institutes of Health (NS109366)

  • Steve Siegelbaum

National Institutes of Health (NS123648)

  • Steve Siegelbaum

Deutsche Forschungsgemeinschaft (FOR 2715 (IS63/10-1/2))

  • Dirk Isbrandt

Deutsche Forschungsgemeinschaft (CRC 1451 (project ID 431549029 - B01))

  • Dirk Isbrandt

Fondazione Telethon (GGP20021)

  • Anna Moroni

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

Reviewing Editor

  1. Dane Michael Chetkovich

Ethics

Animal experimentation: Mouse colonies were maintained both at the University of Cologne and at Columbia University (New York). For research conducted in Cologne, all experiments were in accordance with European, national and institutional guidelines and approved by the State Office of North Rhine-Westphalia, Department of Nature, Environment and Consumer Protection (LANUV NRW, Germany; reference number 81-02.04.2018.A085). For research conducted in New York, all animal experiments were conducted in accordance with policies of the NIH Guide for the Care and Use of Laboratory Animals and the Institutional Animal Care and Use Committee of Columbia University (IACUC protocols AABL5560, AABL5563, AABI2614 and AAAX6450).

Version history

  1. Received: May 31, 2021
  2. Preprint posted: August 17, 2021 (view preprint)
  3. Accepted: August 15, 2022
  4. Accepted Manuscript published: August 16, 2022 (version 1)
  5. Version of Record published: September 16, 2022 (version 2)

Copyright

© 2022, Merseburg 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

  • 1,443
    Page views
  • 338
    Downloads
  • 5
    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. Andrea Merseburg
  2. Jacquelin Kasemir
  3. Eric W Buss
  4. Felix Leroy
  5. Tobias Bock
  6. Alessandro Porro
  7. Anastasia Barnett
  8. Simon E Tröder
  9. Birgit Engeland
  10. Malte Stockebrand
  11. Anna Moroni
  12. Steve Siegelbaum
  13. Dirk Isbrandt
  14. Bina Santoro
(2022)
Seizures, behavioral deficits and adverse drug responses in two new genetic mouse models of HCN1 epileptic encephalopathy
eLife 11:e70826.
https://doi.org/10.7554/eLife.70826

Share this article

https://doi.org/10.7554/eLife.70826

Further reading

    1. Neuroscience
    Sydney Trask, Nicole C Ferrara
    Insight

    Gradually reducing a source of fear during extinction treatments may weaken negative memories in the long term.

    1. Cell Biology
    2. Neuroscience
    Haibin Yu, Dandan Liu ... Kai Yuan
    Research Article

    O-GlcNAcylation is a dynamic post-translational modification that diversifies the proteome. Its dysregulation is associated with neurological disorders that impair cognitive function, and yet identification of phenotype-relevant candidate substrates in a brain-region specific manner remains unfeasible. By combining an O-GlcNAc binding activity derived from Clostridium perfringens OGA (CpOGA) with TurboID proximity labeling in Drosophila, we developed an O-GlcNAcylation profiling tool that translates O-GlcNAc modification into biotin conjugation for tissue-specific candidate substrates enrichment. We mapped the O-GlcNAc interactome in major brain regions of Drosophila and found that components of the translational machinery, particularly ribosomal subunits, were abundantly O-GlcNAcylated in the mushroom body of Drosophila brain. Hypo-O-GlcNAcylation induced by ectopic expression of active CpOGA in the mushroom body decreased local translational activity, leading to olfactory learning deficits that could be rescued by dMyc overexpression-induced increase of protein synthesis. Our study provides a useful tool for future dissection of tissue-specific functions of O-GlcNAcylation in Drosophila, and suggests a possibility that O-GlcNAcylation impacts cognitive function via regulating regional translational activity in the brain.