Canal KCNQ2 mal localizado como mecanismo patogénico en la encefalopatía del desarrollo y epiléptica KCNQ2

KCNQ2 potassium channel variants are linked to developmental and epileptic encephalopathy (DEE). However, the mechanisms by which pathogenic variants, especially those outside known hotspots, such as the S4-S5 linker, lead to disease remain unknown. Here, we examined the H228R variant, a pathogenic mutation in the S4-S5 linker associated with DEE. We tested whether H228R induces KCNQ2 channel mistargeting in addition to its biophysical effects, given recent evidence of impaired trafficking in KCNQ2 DEE variants. We confirmed the H228R variant as a loss-of-function (LOF) when expressed as a homomer and as a dominant-negative when co-expressed with wild-type (WT) KCNQ3. Surprisingly, it exhibited some gain-of-function effects when co-expressed with WT KCNQ2. To determine its cellular localization in vivo, we used male and female heterozygous Kcnq2^H228R knock-in mice, some of which die prematurely despite lack of increases in hippocampal excitatory neuron intrinsic excitability. We validated two different KCNQ2 antibodies in hippocampus via immunohistochemistry. These antibodies detected KCNQ2 in axons, with signal loss observed in Kcnq2 knockout mice. Using these antibodies, we found that the H228R variant caused KCNQ2 channels to concentrate in the soma, strongly reducing their presence in axons. In contrast, analysis of heterozygous mice expressing both a FLAG-tagged WT KCNQ2 and H228R revealed that the FLAG-WT KCNQ2 could still traffic to axons; indicating that some KCNQ2 channels are correctly targeted within neurons. In summary, our results demonstrate that the LOF H228R variant disrupts the localization of variant KCNQ2 channels, suggesting mislocalization as a general endophenotype of KCNQ2 encephalopathy.Significance statement Potassium Voltage-Gated Channel, Subfamily Q, Member 2 (KCNQ2) variants lead to neurodevelopmental disorders. However, the mechanisms by which pathogenic KCNQ2 variants contribute to these conditions remain unclear. In this study, we investigated the KCNQ2 loss-of-function variant H228R, previously identified in a patient with KCNQ2 developmental and epileptic encephalopathy. We found that Kcnq2^H228R heterozygous mice exhibited premature lethality and reduced KCNQ2 protein levels, yet no increases in neuronal intrinsic excitability of hippocampal excitatory neurons. Instead, we observed increased mislocalization of KCNQ2 channels in Kcnq2^H228R heterozygous mice. These results suggest that channel mislocalization may be a common mechanism underlying KCNQ2 loss-of-function variants.