De novo CLPTM1 variants with reduced GABAAR current response in patients with epilepsy

Abstract Objective To investigate the clinical features and potential pathogenesis mechanism of de novo CLPTM1 variants associated with epilepsy. Methods Identify de novo genetic variants associated with epilepsy by reanalyzing trio‐based whole‐exome sequencing data. We analyzed the clinical characteristics of patients with these variants and performed functional in vitro studies in cells expressing mutant complementary DNA for these variants using whole‐cell voltage‐clamp current recordings and outside‐out patch‐clamp recordings from transiently transfected human embryonic kidney (HEK) cells. Results Two de novo missense variants related to epilepsy were identified in the CLPTM1 gene. Functional studies indicated that CLPTM1 ‐p.R454H and CLPTM1 ‐p.R568Q variants reduced the γ‐aminobutyric acid A receptor (GABA A R) current response amplitude recorded under voltage clamp compared to the wild‐type receptors. These variants also reduced the charge transfer and altered the time course of desensitization and deactivation following rapid removal of GABA. The surface expression of the GABA A R γ2 subunit from the CLPTM1 ‐p.R568Q group was significantly reduced compared to CLPTM1 ‐WT. Significance This is the first report of functionally relevant variants within the CLPTM1 gene. Patch‐clamp recordings showed that these de novo CLPTM1 variants reduce GABA A R currents and charge transfer, which should promote excitation and hypersynchronous activity. This study may provide insights into the molecular mechanisms of the CLPTM1 variants underlying the patients’ phenotypes, as well as for exploring potential therapeutic targets for epilepsy.