Developing antisense oligonucleotide therapeutics to target gain-of-function mutations in childhood neurology (360G-Wellcome-215181_Z_19_Z)

The aim of this project is to develop an antisense oligonucleotide (AON) therapeutic approach to target selected gain-of-function mutations that cause childhood neurological disorders. We have selected KCNT1-related infant epilepsy as the primary target disease in this proposal due to the following reasons: 1) this is one of the most frequent genetic causes of infant epilepsies; 2) currently there is no effective treatment; 3) mutations in KCNT1 lead to gain-of-function with a marked, measurable increase in neuronal potassium channel current amplitude; 4) cell lines (fibroblast and induced pluripotent stem cells-iPSCs) are being established from patients and readily quantified functional outcome measures are available. In this study, we will design AONs to selectively target a common KCNT1 mutation for degradation, optimising the most effective through trials of different AON gene-silencing approaches. The effect of AONs on mutant-allele specific silencing will be evaluated in vitro in human cellular models. The phenotypic rescue of lead AONs on potassium channel function will be evaluated in patient’s iPSCs-derived cortical neurons. We expect the success of this project will provide an experimental system and preclinical proof-of-concept of AON therapy applicable for not only KCNT1-related infant epilepsy, but also other childhood neurological disorders caused by gain-of-function mutations.