NaV1.1 and NaV1.6 selective compounds reduce the behavior phenotype and epileptiform activity in a novel zebrafish model for Dravet Syndrome

PLoS ONE, vol. 15, pp. e0219106

Abstract

Dravet syndrome is caused by dominant loss-of-function mutations in SCN1A which cause reduced activity of Nav1.1 leading to lack of neuronal inhibition. On the other hand, gain-of-function mutations in SCN8A can lead to a severe epileptic encephalopathy subtype by over activating NaV1.6 channels. These observations suggest that Nav1.1 and Nav1.6 represent two opposing sides of the neuronal balance between inhibition and activation. Here, we hypothesize that Dravet syndrome may be treated by either enhancing Nav1.1 or reducing Nav1.6 activity. To test this hypothesis we generated and characterized a novel DS zebrafish model and tested new compounds that selectively activate or inhibit the human NaV1.1 or NaV1.6 channel respectively. We used CRISPR/Cas9 to generate two separate Scn1Lab knockout lines as an alternative to previous zebrafish models generated by random mutagenesis or morpholino oligomers. Using an optimized locomotor assay, spontaneous burst movements were detected that were unique to Scn1Lab knockouts and disappear when introducing human SCN1A mRNA. Besides the behavioral phenotype, Scn1Lab knockouts show sudden, electrical discharges in the brain that indicate epileptic seizures in zebrafish. Scn1Lab knockouts showed increased sensitivity to the GABA antagonist pentylenetetrazole and a reduction in whole organism GABA levels. Drug screenings further validated a Dravet syndrome phenotype. We tested the NaV1.1 activator AA43279 and two novel NaV1.6 inhibitors MV1369 and MV1312 in the Scn1Lab knockouts. Both type of compounds significantly reduced the number of spontaneous burst movements and seizure activity. Our results show that selective inhibition of NaV1.6 could be just as efficient as selective activation of NaV1.1 and these approaches could prove to be novel potential treatment strategies for Dravet syndrome and other (genetic) epilepsies. Compounds tested in zebrafish however, should always be further validated in other model systems for efficacy in mammals and to screen for potential side effects.

Authors 12

1. Wout J. Weuring

   Utrecht University · University Medical Center Utrecht

   Affiliation as printed

   Department of Genetics, Center for Molecular Medicine, UMC Utrecht Brain Center, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands

2. Sakshi Singh

   Utrecht University · University Medical Center Utrecht

   Affiliation as printed

   Department of Genetics, Center for Molecular Medicine, UMC Utrecht Brain Center, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands

3. Linda Volkers Aachen Department of Cardiology Laboratory of Experimental Cardiology

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Department of Cardiology, Laboratory of Experimental Cardiology, University Medical Centre Leiden, Leiden, the Netherlands

4. Martin B. Rook

   University Medical Center Utrecht

   Affiliation as printed

   Department of Medical Physiology, University Medical Centre Utrecht, Utrecht, the Netherlands

5. Ruben H. van ’t Slot

   Utrecht University · University Medical Center Utrecht

   Affiliation as printed

   Department of Genetics, Center for Molecular Medicine, UMC Utrecht Brain Center, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands

6. Marjolein Bosma

   Utrecht University · University Medical Center Utrecht

   Affiliation as printed

   Department of Genetics, Center for Molecular Medicine, UMC Utrecht Brain Center, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands

7. Marco C. Inserra

   The University of Queensland

   Affiliation as printed

   Centre for Pain Research & School of Pharmacy, University of Queensland, Brisbane, Australia

8. Irina Vetter

   The University of Queensland

   Affiliation as printed

   Centre for Pain Research & School of Pharmacy, University of Queensland, Brisbane, Australia

9. Nanda M. Verhoeven‐Duif

   Utrecht University · University Medical Center Utrecht

   Affiliation as printed

   Department of Genetics, Center for Molecular Medicine, UMC Utrecht Brain Center, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands

10. Kees P. J. Braun

    University Medical Center Utrecht

    Affiliation as printed

    Department of Neurology, University Medical Centre Utrecht, Utrecht, the Netherlands

11. Mirko Rivara

    University of Parma

    Affiliation as printed

    Food and Drug Department, University of Parma, Parma, Italy

12. Bobby P.C. Koeleman corresponding

    Utrecht University · University Medical Center Utrecht

    Affiliation as printed

    Department of Genetics, Center for Molecular Medicine, UMC Utrecht Brain Center, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands