Viral vector-mediated SLC9A6 gene replacement reduces cerebellar dysfunction in the shaker rat model of Christianson syndrome

Abstract

BackgroundChristianson syndrome (CS) is an x-linked recessive neurodevelopmental and neurodegenerative condition characterized by severe intellectual disability, cerebellar degeneration, ataxia, and epilepsy. Mutations to the SLC9A6 gene encoding NHE6 are responsible for CS, and we recently demonstrated that a mutation to the rat Slc9a6 gene causes a similar phenotype in the spontaneous shaker rat model, which exhibits cerebellar degeneration with motor dysfunction. In previous work, we used the PhP.eB-L7-Slc9a6-GFP adeno-associated viral (AAV) vector to demonstrate that gene replacement in Purkinje cells reduced the shaker motor and molecular phenotype. MethodsWe carried out a 20-week longitudinal study evaluating the impact of Purkinje cell-specific gene replacement on ataxia and tremor. Taking advantage of the high homology between human SLC9A6 and rat Slc9a6, we tested a more clinically relevant construct, AAV9-CAG-hSLC9A6 AAV vector in the shaker rat. In both experimental cohorts, we performed molecular studies to evaluate expression of NHE6 and key cerebellar markers. We then characterized the relationship between molecular markers and motor function, as well between tremor and ataxia. ResultsAdministration of either of PhP.eB-L7-Slc9a6-GFP or AAV9-CAG-hSLC9A6 AAV vectors led to significant improvement in the molecular and motor phenotypes. The abundance of each disease-relevant cerebellar proteins was significantly correlated to motor ataxia. Further, we found that the relationship between cerebellar ataxia and tremor devolved over time, with disease modifying therapy disrupting their temporal relationship. ConclusionsThese findings impact future SLC9A6-targeted gene therapy efforts for CS and strongly support gene replacement as a viable therapeutic strategy. Furthermore, tremor and ataxia phenotypes may arise from dissociable cerebellar mechanisms. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=69 SRC="FIGDIR/small/621435v2_ufig1.gif" ALT="Figure 1"> View larger version (22K): org.highwire.dtl.DTLVardef@368165org.highwire.dtl.DTLVardef@691b54org.highwire.dtl.DTLVardef@816174org.highwire.dtl.DTLVardef@2508fb_HPS_FORMAT_FIGEXP M_FIG C_FIG