Complex biophysical changes and reduced neuronal firing in an SCN8A variant associated with developmental delay and epilepsy

Shir Quinn, Nan Zhang, Timothy A. Fenton, Marina Brusel, Preethi Muruganandam, Yoav Peleg, Moshe Giladi, Yoni Haitin, Holger Lerche, Haim Bassan, Yuanyuan Liu*, Roy Ben-Shalom, Moran Rubinstein*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Mutations in the SCN8A gene, encoding the voltage-gated sodium channel NaV1.6, are associated with a range of neurodevelopmental syndromes. The p.(Gly1625Arg) (G1625R) mutation was identified in a patient diagnosed with developmental epileptic encephalopathy (DEE). While most of the characterized DEE-associated SCN8A mutations were shown to cause a gain-of-channel function, we show that the G1625R variant, positioned within the S4 segment of domain IV, results in complex effects. Voltage-clamp analyses of NaV1.6G1625R demonstrated a mixture of gain- and loss-of-function properties, including reduced current amplitudes, increased time constant of fast voltage-dependent inactivation, a depolarizing shift in the voltage dependence of activation and inactivation, and increased channel availability with high-frequency repeated depolarization. Current-clamp analyses in transfected cultured neurons revealed that these biophysical properties caused a marked reduction in the number of action potentials when firing was driven by the transfected mutant NaV1.6. Accordingly, computational modeling of mature cortical neurons demonstrated a mild decrease in neuronal firing when mimicking the patients' heterozygous SCN8A expression. Structural modeling of NaV1.6G1625R suggested the formation of a cation-π interaction between R1625 and F1588 within domain IV. Double-mutant cycle analysis revealed that this interaction affects the voltage dependence of inactivation in NaV1.6G1625R. Together, our studies demonstrate that the G1625R variant leads to a complex combination of gain and loss of function biophysical changes that result in an overall mild reduction in neuronal firing, related to the perturbed interaction network within the voltage sensor domain, necessitating personalized multi-tiered analysis for SCN8A mutations for optimal treatment selection.

Original languageEnglish
Article number167127
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Volume1870
Issue number5
DOIs
StatePublished - Jun 2024

Funding

FundersFunder number
Kahn Foundation
Tel Aviv Sourasky Medical Center
Hartwell Foundation
Claire and Amedee Maratier Institute for the Study of Blindness
Weizmann Institute of Science
Stolz Foundation Faculty of Medicine, Tel Aviv University
Tel Aviv University
University of TuebingenN15/18 M
Bundesministerium für Bildung und Forschung01GM2210A
Bundesministerium für Bildung und Forschung
Israel Science Foundation1653/21, 1454/17, 214/22
Israel Science Foundation
Israel Cancer Research Fund19202
Israel Cancer Research Fund
Deutsche ForschungsgemeinschaftLe1030/15-2
Deutsche Forschungsgemeinschaft
Israel Cancer Association20230029
Israel Cancer Association

    Keywords

    • Biophysical analysis
    • Developmental epileptic encephalopathy
    • G1625R
    • Neuronal and structural modeling
    • Patch-clamp
    • SCN8A

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