Abstract
Gene therapy offers a potential alternative to the surgical treatment of epilepsy, which affects millions of people and is pharmacoresistant in ~30% of cases. Aimed at reducing the excitability of principal neurons, the engineered expression of K+ channels has been proposed as a treatment due to the outstanding ability of K+ channels to hyperpolarize neurons. However, the effects of K+ channel overexpression on cell physiology remain to be investigated. Here we report an adeno-associated virus (AAV) vector designed to reduce epileptiform activity specifically in excitatory pyramidal neurons by expressing the human Ca2+-gated K+ channel KCNN4 (KCa3.1). Electrophysiological and pharmacological experiments in acute brain slices showed that KCNN4-transduced cells exhibited a Ca2+-dependent slow afterhyperpolarization that significantly decreased the ability of KCNN4-positive neurons to generate high-frequency spike trains without affecting their lower-frequency coding ability and action potential shapes. Antiepileptic activity tests showed potent suppression of pharmacologically induced seizures in vitro at both single cell and local field potential levels with decreased spiking during ictal discharges. Taken together, our findings strongly suggest that the AAV-based expression of the KCNN4 channel in excitatory neurons is a promising therapeutic intervention as gene therapy for epilepsy.
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All relevant data supporting the key findings of this study are available within the article or from corresponding authors on request.
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Acknowledgements
This research was funded by Russian Science Foundation; grant: RSF 20-15-00408. We would like to thank Dr. Alexander S. Chernyshev for his help in analyzing the field recordings.
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Conceptualization, ESN, VVB, PMB, and AVZ; Funding acquisition, PMB; ESN, MVR, TYP, VI, IK, AAB, EYP, MPS, and AVZ performed the experiments; Methodology, AAB, ESN, TYP, and AVZ; Project administration, ESN, PMB, and AVZ; Writing—original draft, ESN, TYP, EYP, and AVZ; Reviewing & editing, ESN, PMB, VVB, and AVZ. All authors have read and agreed to the published version of the manuscript.
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Nikitin, E.S., Postnikova, T.Y., Proskurina, E.Y. et al. Overexpression of KCNN4 channels in principal neurons produces an anti-seizure effect without reducing their coding ability. Gene Ther 31, 144–153 (2024). https://doi.org/10.1038/s41434-023-00427-9
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DOI: https://doi.org/10.1038/s41434-023-00427-9
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