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 4

P21 injection with ASO successfully restores UBE3A expression in AS mice and rescues epilepsy and hippocampal plasticity.

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P21 injection with ASO successfully restores UBE3A expression in AS mice...
(A) Schematic representation of the timeline for ASO treatment, epilepsy testing, and LTP measurement. (B) Three weeks postinjection (n = 2) the brains of WT-PBS, AS-ASO, and AS-PBS mice were stained for UBE3A (green). The brains of AS mice treated with ASO show widely distributed nuclear UBE3A compared with the untreated AS brain. (C–F) Western blots of lysates obtained 4 weeks after P21 injection of cortex, hippocampus, striatum, and cerebellum of AS mice treated with ASO (n = 3) and compared with age-matched WT controls and AS mice injected with PBS (n = 3). Two bands, representing the cytosolic and nuclear isoforms of UBE3A, can be detected at 100 kDa and ACTIN, here used as loading control, at 45 kDa. One-way ANOVA, followed by Tukey’s post hoc test, was used as statistical test. (G) Susceptibility to seizures visually observed in AS mice (n = 10) is rescued when treated with ASO at P21 (n = 13). Seizures are also absent in WT mice (n = 9). Fisher exact test was used as statistical test. (H) LTP measurement in AS mice treated with ASO (blue line), WT mice (black line), and AS mice (red line). Two-way ANOVA, followed by Bonferroni’s correction, was used as the statistical test. Number of slices used/mouse: AS-PBS (n = 26/5); AS-ASO (n = 27/7); WT-PBS (n = 23/4). Scale bars: 200 μm (B). Data are represented as means ± 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.