Abstract

Deafness non-syndromic autosomal dominant 2 (DFNA2) is characterized by symmetric, predominantly high-frequency sensorineural hearing loss that is progressive across all frequencies. The disease is associated with variants of a potassium voltage-gated channel subfamily Q member 4 gene, KCNQ4 (Kv7.4). Here, we studied nine recently identified Kv7.4 variants in DFNA2 pedigrees, including V230E, E260K, D262V, Y270H, W275R, G287R, P291L, P291S and S680F. We proved that the variant S680F did not alter the channel function while the other eight variants resulted in function deficiencies. We further proved that the two variants E260K and P291S showed reduced cell membrane expressions while the other seven variants showed moderate cell surface expressions. Thus, trafficking deficiency is not a common mechanism underlying channel dysfunction. Next, we studied two variants, V230E and G287R, using molecular dynamics simulation. We showed that V230E stabilized Kv7.4 channel in the closed state by forming an additional hydrogen bond with a basic residue K325, while G287R distorted the selectivity filter and blocked the pore region of Kv7.4 channel. Moreover, by co-expressing wild-type (WT) and variant proteins in vitro , we demonstrated that the heterogeneous Kv7.4 channel currents were reduced compared to the WT channel currents and the reduction could be rescued by a Kv7.4 opener retigabine . Our study provided the underlying mechanisms and suggested a potential alternative therapeutic approach for DFNA2.