LAUSR

OBJECTIVE Pathogenic variants in KCNT2 are rare causes of developmental epileptic encephalopathy (DEE). We herein describe the phenotypic and genetic features of patients with KCNT2-related DEE, and the in vitro functional and pharmacological properties of KCNT2 channels carrying fourteen novel or previously untested variants. METHODS 25 patients harbouring KCNT2 variants were investigated: 12 were identified through an international collaborative network, 13 were retrieved from the literature. Clinical data were collected and included in a standardized phenotyping sheet. Novel variants were detected using exome sequencing and classified using ACMG criteria. Functional and pharmacological studies were performed by whole-cell electrophysiology in HEK-293 cells. RESULTS The phenotypic spectrum encompassed: a) Intellectual disability/Developmental delay (21/22 individuals with available information), ranging from mild to severe/profound; b) epilepsy (15/25); c) neurological impairment, with altered muscle tone (14/22); d) dysmorphisms (13/20). Nineteen pathogenic KCNT2 variants were found (nine new, ten reported previously): 16 missense, one in-frame deletion of a single amino acid, one nonsense, and one frameshift. Among tested variants, eight showed gain-of-function (GoF), and six loss-of-function (LoF) features when expressed heterologously in vitro. Quinidine and fluoxetine blocked all GoF variants, whereas loxapine and riluzole activated some LoF variants while blocking others. INTERPRETATION We expanded the phenotypic and genotypic spectrum of KCNT2-related disorders, highlighting novel genotype-phenotype associations. Pathogenic KCNT2 variants cause GoF or LoF in vitro phenotypes, and each shows a unique pharmacological profile, suggesting the need for in vitro functional and pharmacological investigation to enable targeted therapies based on the molecular phenotype. This article is protected by copyright. All rights reserved.