Prax330 reduces persistent and resurgent sodium channel currents and neuronal hyperexcitability of subiculum neurons in a mouse model of SCN8A epileptic encephalopathy

摘要

SCN8A epileptic encephalopathy is a severe genetic epilepsy syndrome caused by de novo gain-of-function mutations of SCN8A encoding the voltage-gated sodium (Na) channel (VGSC) Na(v)1.6. Therapeutic management is difficult in many patients, leading to uncontrolled seizures and risk of sudden unexpected death in epilepsy (SUDEP). There is a need to develop novel anticonvulsants that can specifically target aberrant VGSC activity associated with SCN8A gain-of-function mutations. In this study, we investigate the effects of Prax330, a novel VGSC inhibitor, on the biophysical properties of wild-type (WT) Na(v)1.6 and the patient mutation p.Asn1768Asp (N1768D) in ND7/23 cells. The effects of Prax330 on persistent (I-Nap) and resurgent (I-NaR) Na currents and neuronal excitability in subiculum neurons from a knock-in mouse model of the Scn8 alpha-N1768D mutation (Scn8 alpha(D/+)) were also examined. In ND7/23 cells, Prax330 reduced I-Nap currents recorded from cells expressing Scn8 alpha-N1768D and hyperpolarized steady-state inactivation curves. Recordings from brain slices demonstrated elevated I-Nap and I-NaR in subiculum neurons from Scn8 alpha(D/+) mutant mice and abnormally large action potential (AP) burst-firing events in a subset of neurons. Prax330 (1 mu M) reduced both I-Nap and I-NaR and suppressed AP bursts, with a smaller effect on AP waveforms that had similar morphology to WT neurons. Prax330 (1 mu M) also reduced synaptically-evoked APs in Scn8 alpha(D/+) subiculum neurons but not in WT neurons. Our results highlight the efficacy of targeting I-Nap and I-NaR and inactivation parameters in controlling subiculum excitability and suggest Prax330 as a promising novel therapy for SCN8A epileptic encephalopathy.