Viral vector-mediated SLC9A6 gene replacement reduces cerebellar motor and molecular abnormalities in the shaker rat model of Christianson syndrome

Abstract Background Christianson syndrome is an x-linked recessive neurodevelopmental and neurodegenerative condition caused by mutations to the SLC9A6 gene encoding NHE6, a sodium-hydrogen exchanger critical for regulating endosomal pH. Using an adeno-associated viral (AAV) vector targeting Purkinje cells (PHP.eB-L7-Slc9a6-GFP), we recently performed functional complementation studies to demonstrate that mutation to the rat Slc9a6 gene causes a Christianson syndrome-relevant cerebellar phenotype in the shaker rat. Methods We carried out a longitudinal study evaluating the impact of gene replacement targeting Purkinje cells on ataxia and tremor in the shaker rat. Further, in a smaller follow-up study, we tested administration of AAV9-CAG-hSLC9A6 to determine whether key molecular and motor findings could be replicated with a more clinically relevant viral construct. In both experimental cohorts, we performed molecular studies to evaluate expression of NHE6 and key cerebellar markers. Results Administration of either of PHP.eB-L7-Slc9a6-GFP or AAV9-CAG-hSLC9A6 AAV vectors led to significant improvement in both the molecular and motor phenotypes. The abundance of each disease-relevant cerebellar proteins was strongly correlated to motor ataxia, but less so to tremor. Further, while ataxia and tremor were initially strongly correlated early in the disease progression, this relationship weakened over time. Conclusions These findings impact future SLC9A6-targeted gene therapy efforts for Christianson syndrome and support gene replacement as a potentially viable therapeutic strategy. Common markers associated with cerebellar degeneration are much more strongly tied to ataxia than to tremor, indicating that ataxia may be more tied related to degenerative processes than tremor.