Antisense oligonucleotide treatment rescues UBE3A expression and multiple phenotypes of an Angelman syndrome mouse model
Claudia Milazzo, Edwin J. Mientjes, Ilse Wallaard, Søren Vestergaard Rasmussen, Kamille Dumong Erichsen, Tejaswini Kakunuri, A.S. Elise van der Sman, Thomas Kremer, Meghan T. Miller, Marius C. Hoener, Ype Elgersma
Claudia Milazzo, Edwin J. Mientjes, Ilse Wallaard, Søren Vestergaard Rasmussen, Kamille Dumong Erichsen, Tejaswini Kakunuri, A.S. Elise van der Sman, Thomas Kremer, Meghan T. Miller, Marius C. Hoener, Ype Elgersma
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Research Article Development Neuroscience

Antisense oligonucleotide treatment rescues UBE3A expression and multiple phenotypes of an Angelman syndrome mouse model

Abstract

Angelman syndrome (AS) is a severe neurodevelopmental disorder for which only symptomatic treatment with limited benefits is available. AS is caused by mutations affecting the maternally inherited ubiquitin protein ligase E3A (UBE3A) gene. Previous studies showed that the silenced paternal Ube3a gene can be activated by targeting the antisense Ube3a-ATS transcript. We investigated antisense oligonucleotide–induced (ASO-induced) Ube3a-ATS degradation and its ability to induce UBE3A reinstatement and rescue of AS phenotypes in an established Ube3a mouse model. We found that a single intracerebroventricular injection of ASOs at postnatal day 1 (P1) or P21 in AS mice resulted in potent and specific UBE3A reinstatement in the brain, with levels up to 74% of WT levels in the cortex and a full rescue of sensitivity to audiogenic seizures. AS mice treated with ASO at P1 also showed rescue of established AS phenotypes, such as open field and forced swim test behaviors, and significant improvement on the reversed rotarod. Hippocampal plasticity of treated AS mice was comparable to WT but not significantly different from PBS-treated AS mice. No rescue was observed for the marble burying and nest building phenotypes. Our findings highlight the promise of ASO-mediated reactivation of UBE3A as a disease-modifying treatment for AS.

Authors

Claudia Milazzo, Edwin J. Mientjes, Ilse Wallaard, Søren Vestergaard Rasmussen, Kamille Dumong Erichsen, Tejaswini Kakunuri, A.S. Elise van der Sman, Thomas Kremer, Meghan T. Miller, Marius C. Hoener, Ype Elgersma

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Figure 3

ICV injection of ASO at P1 rescues specific AS phenotypes.

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ICV injection of ASO at P1 rescues specific AS phenotypes. (A) Schematic...
(A) Schematic representation of ASO treatment and timeline for behavioral testing. (B–F) Behavioral battery performed with WT-PBS (black, n = 14), AS-PBS (red, n = 16), and AS mice injected with ASO (blue, n = 16) in the open field, marble burying, nest building, forced swim tests, and accelerating rotarod. One-way or 2-way ANOVA was performed, with treatment plus genotype as independent variable for statistical analysis. (G) A new cohort of 6-week-old AS-PBS (n = 30), WT-PBS (n = 19), and AS-ASO (n = 17) mice was used for the reverse rotarod. A 2-way ANOVA was performed, with treatment plus genotype as independent variable for statistical analysis. (H) Brain weight of 7-week-old AS-PBS (n = 7), WT-PBS (n = 8), and AS-ASO (n = 8) mice. One-way ANOVA was performed, with treatment plus genotype as independent variable for statistical analysis. (I) Audiogenic seizures were rescued in P1-ASO–treated Ube3a–/+ mice: susceptibility to clonic-tonic seizures was visually observed in AS mice (n = 6) and not observed in ASO-treated mice (n = 10) or WT mice (n = 7). Fisher exact test was used as the statistical test. (J) LTP in AS mice treated with ASO showed a trend similar to WT mice. Both gave a higher response than AS mice, but this difference was not significant. Two-way ANOVA, followed by Bonferroni’s post hoc test, was used as the statistical test, with treatment and genotype as independent variables. Number of slices/mouse measured: AS-PBS (n = 13/6); AS-ASO (n = 16/8); WT-PBS (n = 13/6). Data represented as mean ± SEM. P values represent the significance level for the parameter and are displayed as stars in the figure: not shown if P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. fEPSP, field excitatory postsynaptic potential.