Antisense oligonucleotides restore excitability, GABA signalling and sodium current density in a Dravet syndrome model

Brain. 2024 Apr 4;147(4):1231-1246. doi: 10.1093/brain/awad349.

Abstract

Dravet syndrome is an intractable developmental and epileptic encephalopathy caused by de novo variants in SCN1A resulting in haploinsufficiency of the voltage-gated sodium channel Nav1.1. We showed previously that administration of the antisense oligonucleotide STK-001, also called ASO-22, generated using targeted augmentation of nuclear gene output technology to prevent inclusion of the nonsense-mediated decay, or poison, exon 20N in human SCN1A, increased productive Scn1a transcript and Nav1.1 expression and reduced the incidence of electrographic seizures and sudden unexpected death in epilepsy in a mouse model of Dravet syndrome. Here, we investigated the mechanism of action of ASO-84, a surrogate for ASO-22 that also targets splicing of SCN1A exon 20N, in Scn1a+/- Dravet syndrome mouse brain. Scn1a +/- Dravet syndrome and wild-type mice received a single intracerebroventricular injection of antisense oligonucleotide or vehicle at postnatal Day 2. We examined the electrophysiological properties of cortical pyramidal neurons and parvalbumin-positive fast-spiking interneurons in brain slices at postnatal Days 21-25 and measured sodium currents in parvalbumin-positive interneurons acutely dissociated from postnatal Day 21-25 brain slices. We show that, in untreated Dravet syndrome mice, intrinsic cortical pyramidal neuron excitability was unchanged while cortical parvalbumin-positive interneurons showed biphasic excitability with initial hyperexcitability followed by hypoexcitability and depolarization block. Dravet syndrome parvalbumin-positive interneuron sodium current density was decreased compared to wild-type. GABAergic signalling to cortical pyramidal neurons was reduced in Dravet syndrome mice, suggesting decreased GABA release from interneurons. ASO-84 treatment restored action potential firing, sodium current density and GABAergic signalling in Dravet syndrome parvalbumin-positive interneurons. Our work suggests that interneuron excitability is selectively affected by ASO-84. This new work provides critical insights into the mechanism of action of this antisense oligonucleotide and supports the potential of antisense oligonucleotide-mediated upregulation of Nav1.1 as a successful strategy to treat Dravet syndrome.

Keywords: antisense oligonucleotide; epilepsy; sodium channel; therapeutics.

MeSH terms

  • Animals
  • Disease Models, Animal
  • Epilepsies, Myoclonic* / genetics
  • Humans
  • Interneurons / metabolism
  • Mice
  • NAV1.1 Voltage-Gated Sodium Channel / genetics
  • Oligonucleotides, Antisense* / pharmacology
  • Parvalbumins / metabolism
  • gamma-Aminobutyric Acid

Substances

  • Oligonucleotides, Antisense
  • Parvalbumins
  • NAV1.1 Voltage-Gated Sodium Channel
  • gamma-Aminobutyric Acid
  • Scn1a protein, mouse