Interneuron-specific dual-AAV SCN1A gene replacement corrects epileptic phenotypes in mouse models of Dravet syndrome

Sci Transl Med. 2025 Mar 19;17(790):eadn5603. doi: 10.1126/scitranslmed.adn5603. Epub 2025 Mar 19.

Abstract

Dravet syndrome (DS) is a severe developmental epileptic encephalopathy marked by treatment-resistant seizures, developmental delay, intellectual disability, motor deficits, and a 10 to 20% rate of premature death. Most patients with DS harbor loss-of-function mutations in one copy of SCN1A, which encodes the voltage-gated sodium channel (NaV)1.1 alpha subunit and has been associated with inhibitory neuron dysfunction. Here, we generated a split-intein form of SCN1A and used a dual-vector delivery approach to circumvent adeno-associated virus (AAV) packaging limitations. In addition, we applied previously developed enhancer technology to produce an interneuron-specific gene replacement therapy for DS, called DLX2.0-SCN1A. The split-intein SCN1A vectors produced full-length NaV1.1 protein, and functional sodium channels were recorded in HEK293 cells in vitro. Administration of dual DLX2.0-SCN1A AAVs to wild-type mice produced full-length, reconstituted human protein by Western blot and telencephalic interneuron-specific and dose-dependent NaV1.1 expression by immunohistochemistry. These vectors also conferred strong dose-dependent protection against postnatal mortality and seizures in Scn1afl/+;Meox2-Cre and Scn1a+/R613X DS mouse models. Injection of single or dual DLX2.0-SCN1A AAVs into wild-type mice did not result in increased mortality, weight loss, or gliosis as measured by immunohistochemistry. In contrast, expression of SCN1A in all neurons driven by the human SYNAPSIN I promoter caused an adverse effect marked by increased mortality in the preweaning period, before disease onset. These findings demonstrate proof of concept that interneuron-specific AAV-mediated SCN1A gene replacement can rescue DS phenotypes in mouse models and suggest that it could be a therapeutic approach for patients with DS.

MeSH terms

  • Animals
  • Dependovirus* / genetics
  • Disease Models, Animal
  • Epilepsies, Myoclonic* / genetics
  • Epilepsies, Myoclonic* / pathology
  • Epilepsies, Myoclonic* / therapy
  • Genetic Therapy* / methods
  • Genetic Vectors
  • HEK293 Cells
  • Humans
  • Interneurons* / metabolism
  • Mice
  • NAV1.1 Voltage-Gated Sodium Channel* / genetics
  • NAV1.1 Voltage-Gated Sodium Channel* / metabolism
  • Phenotype

Substances

  • NAV1.1 Voltage-Gated Sodium Channel
  • Scn1a protein, mouse
  • SCN1A protein, human