Discovery and characterization of a specific inhibitor of serine-threonine kinase cyclin dependent kinase-like 5 (CDKL5) demonstrates role in hippocampal CA1 physiology

  1. Anna Castano
  2. Margaux Silvestre
  3. Carrow I Wells
  4. Jennifer L Sanderson
  5. Carla A Ferrer
  6. Han Wee Ong
  7. Yi Lang
  8. William Richardson
  9. Josie A Silvaroli
  10. Frances M Bashore
  11. Jeffery L Smith
  12. Isabelle M Genereux
  13. Kelvin Dempster
  14. David H Drewry
  15. Navlot S Pabla
  16. Alex N Bullock
  17. Tim A Benke  Is a corresponding author
  18. Sila Ultanir  Is a corresponding author
  19. Alison D Axtman  Is a corresponding author
  1. University of Colorado Anschutz Medical Campus, United States
  2. The Francis Crick Institute, United Kingdom
  3. University of North Carolina at Chapel Hill, United States
  4. University of Oxford, United Kingdom
  5. The Ohio State University, United States

Abstract

Pathological loss-of-function mutations in cyclin-dependent kinase-like 5 (CDKL5) cause CDKL5 deficiency disorder (CDD), a rare and severe neurodevelopmental disorder associated with severe and medically refractory early-life epilepsy, motor, cognitive, visual and autonomic disturbances in the absence of any structural brain pathology. Analysis of genetic variants in CDD have indicated that CDKL5 kinase function is central to disease pathology. CDKL5 encodes a serine-threonine kinase with significant homology to GSK3b, which has also been linked to synaptic function. Further, Cdkl5 knock-out rodents have increased GSK3b activity and often increased long-term potentiation (LTP). Thus, development of a specific CDKL5 inhibitor must be careful to exclude cross-talk with GSK3b activity. We synthesized and characterized specific, high-affinity inhibitors of CDKL5 that do not have detectable activity for GSK3b. These compounds are very soluble in water but blood-brain barrier penetration is low. In rat hippocampal brain slices, acute inhibition of CDKL5 selectively reduces post-synaptic function of AMPA-type glutamate receptors in a dose-dependent manner. Acute inhibition of CDKL5 reduces hippocampal LTP. These studies provide new tools and insights into the role of CDKL5 as a newly appreciated, key kinase necessary for synaptic plasticity. Comparisons to rodent knock-out studies suggest that compensatory changes have limited the understanding of the roles of CDKL5 in synaptic physiology, plasticity and human neuropathology.

Data availability

Materials and data availability statement has been provided in the text: Data are available via Dryad (https://doi.org/10.5061/dryad.sn02v6x88). New reagents are available from the senior authors.

The following data sets were generated

Article and author information

Author details

  1. Anna Castano

    Department of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, United States
    Competing interests
    No competing interests declared.
  2. Margaux Silvestre

    Kinases and Brain Development Laboratory, The Francis Crick Institute, London, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1377-477X
  3. Carrow I Wells

    Structural Genomics Consortium, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4799-6792
  4. Jennifer L Sanderson

    Department of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, United States
    Competing interests
    No competing interests declared.
  5. Carla A Ferrer

    Structural Genomics Consortium, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    No competing interests declared.
  6. Han Wee Ong

    Structural Genomics Consortium, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    No competing interests declared.
  7. Yi Lang

    Structural Genomics Consortium, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    No competing interests declared.
  8. William Richardson

    Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  9. Josie A Silvaroli

    Division of Pharmaceutics and Pharmacology, The Ohio State University, Columbus, United States
    Competing interests
    No competing interests declared.
  10. Frances M Bashore

    Structural Genomics Consortium, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4241-9873
  11. Jeffery L Smith

    Structural Genomics Consortium, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2189-0420
  12. Isabelle M Genereux

    Structural Genomics Consortium, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    No competing interests declared.
  13. Kelvin Dempster

    Kinases and Brain Development Laboratory, The Francis Crick Institute, London, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0009-0009-0750-0175
  14. David H Drewry

    Structural Genomics Consortium, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    No competing interests declared.
  15. Navlot S Pabla

    Division of Pharmaceutics and Pharmacolog, The Ohio State University, Columbu, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9408-0539
  16. Alex N Bullock

    Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  17. Tim A Benke

    Department of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, United States
    For correspondence
    tim.benke@cuanschutz.edu
    Competing interests
    Tim A Benke, Consultancy for AveXis, Ovid, GW Pharmaceuticals, International Rett Syndrome Foundation, Takeda, Taysha, CureGRIN, GRIN Therapeutics, Alcyone, Neurogene, and Marinus; Clinical Trials with Acadia, Ovid, GW Pharmaceuticals, Marinus and RSRT; all remuneration has been made to his department..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6969-5061
  18. Sila Ultanir

    Kinases and Brain Development Laboratory, The Francis Crick Institute, London, United Kingdom
    For correspondence
    sila.ultanir@crick.ac.uk
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5745-3957
  19. Alison D Axtman

    Structural Genomics Consortium, University of North Carolina at Chapel Hill, Chapel Hill, United States
    For correspondence
    Alison.Axtman@unc.edu
    Competing interests
    Alison D Axtman, Advisor for Proteic Bioscience Inc..

Funding

National Institute of Neurological Disorders and Stroke (NS112770)

  • Anna Castano
  • Margaux Silvestre
  • Carrow I Wells
  • Jennifer L Sanderson
  • Carla A Ferrer
  • Han Wee Ong
  • Yi Lang
  • William Richardson
  • Josie A Silvaroli
  • Frances M Bashore
  • Jeffery L Smith
  • Isabelle M Genereux
  • Kelvin Dempster
  • David H Drewry
  • Navlot S Pabla
  • Alex N Bullock
  • Tim A Benke
  • Sila Ultanir
  • Alison D Axtman

LouLou Foundation (11015)

  • Margaux Silvestre

Children's Hospital Colorado Foundation (Ponzio Family Chair in Neuroscience Research)

  • Anna Castano
  • Jennifer L Sanderson
  • Tim A Benke

Structural Genomics Consortium

  • Carrow I Wells
  • Carla A Ferrer
  • Han Wee Ong
  • Yi Lang
  • Frances M Bashore
  • Jeffery L Smith
  • Isabelle M Genereux
  • David H Drewry
  • Alison D Axtman

NC Biotechnology Center Institutional Support Grant (2018-IDG-1030)

  • David H Drewry
  • Alison D Axtman

National Institutes of Health (U24DK116204)

  • David H Drewry
  • Alison D Axtman

National Institutes of Health (R44TR001916)

  • David H Drewry
  • Alison D Axtman

Cancer Research UK (CC2037)

  • Margaux Silvestre
  • Sila Ultanir

Medical Research Council (CC2037)

  • Margaux Silvestre
  • Sila Ultanir

Wellcome Trust (CC2037)

  • Margaux Silvestre
  • Sila Ultanir

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

Reviewing Editor

  1. Alan Talevi, National University of La Plata, Argentina

Ethics

Animal experimentation: Animals-The Francis Crick Institute (TFCI). Rat handling and housing was performed according to the regulations of the Animal (Scientific Procedures) Act 1986. Animal studies were approved by the Francis Crick Institute ethical committee and performed under U.K. Home Office project license (PPL P5E6B5A4B).Animals-University of Colorado School of Medicine (UC-SOM). All studies conformed to the requirements of the National Institutes of Health Guide for the Care and Use of Laboratory Rats and were approved by the Institutional Animal Care and Use subcommittee of the University of Colorado Anschutz Medical Campus (protocol 00411). Timed-pregnant Sprague Dawley rats (Charles Rivers Labs, Wilmington, MA, USA) gave birth in-house. All rodents were housed in micro-isolator cages with water and chow available ad libitum.

Version history

  1. Received: March 29, 2023
  2. Preprint posted: April 24, 2023 (view preprint)
  3. Accepted: July 24, 2023
  4. Accepted Manuscript published: July 25, 2023 (version 1)
  5. Version of Record published: August 7, 2023 (version 2)

Copyright

© 2023, Castano 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.

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  1. Anna Castano
  2. Margaux Silvestre
  3. Carrow I Wells
  4. Jennifer L Sanderson
  5. Carla A Ferrer
  6. Han Wee Ong
  7. Yi Lang
  8. William Richardson
  9. Josie A Silvaroli
  10. Frances M Bashore
  11. Jeffery L Smith
  12. Isabelle M Genereux
  13. Kelvin Dempster
  14. David H Drewry
  15. Navlot S Pabla
  16. Alex N Bullock
  17. Tim A Benke
  18. Sila Ultanir
  19. Alison D Axtman
(2023)
Discovery and characterization of a specific inhibitor of serine-threonine kinase cyclin dependent kinase-like 5 (CDKL5) demonstrates role in hippocampal CA1 physiology
eLife 12:e88206.
https://doi.org/10.7554/eLife.88206

Share this article

https://doi.org/10.7554/eLife.88206

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