Investigation of novel de novo KCNC2 variants causing severe developmental and early-onset epileptic encephalopathy

Purpose The voltage-gated potassium channel K(v)3.2, encoded by KCNC2, facilitates fast-spiking GABAergic interneurons to fire action potentials at high frequencies. It is pivotal to maintaining excitation/inhibition balance in mammalian brains. This study identified two novel de novo KCNC2 variants, p.Pro470Ser (P470S) and p. Phe382Leu (F382L), in patients with early onset developmental and epileptic encephalopathy (DEE). Methods To examine the molecular basis of DEE, we studied the functional characteristics of variant channels using patch-clamp techniques and computational modeling. Results Whole-cell patch clamp recordings from infected HEK293 cells revealed that channel activation and deactivation kinetics strongly decreased in both K(v)3.2 P470S and F382L variant channels. This decrease also occurred in K(v)3.2 p.Val471Leu (V471L) channels, known to be associated with DEE. In addition, K(v)3.2 F382L and V471L variants exhibited a significant increase in channel conductance and a similar to 20 mV negative shift in the threshold for voltage-dependent activation. Simulations of model GABAergic interneurons revealed that all variants decreased neuronal firing frequency. Thus, the variants' net loss-of-function effects disinhibited neural networks. Conclusion Our findings provide compelling evidence supporting the role of KCNC2 as a disease-causing gene in human neurodevelopmental delay and epilepsy.

Automated summary

This study identified two novel KCNC2 variants in patients with early onset developmental and epileptic encephalopathy, providing evidence supporting the role of KCNC2 as a disease-causing gene in human neurodevelopmental delay and epilepsy.