Dual-isoform hUBE3A gene transfer improves behavioral and seizure outcomes in Angelman syndrome model mice
Matthew C. Judson, Charles Shyng, Jeremy M. Simon, Courtney R. Davis, A. Mattijs Punt, Mirabel T. Salmon, Noah W. Miller, Kimberly D. Ritola, Ype Elgersma, David G. Amaral, Steven J. Gray, Benjamin D. Philpot
Matthew C. Judson, Charles Shyng, Jeremy M. Simon, Courtney R. Davis, A. Mattijs Punt, Mirabel T. Salmon, Noah W. Miller, Kimberly D. Ritola, Ype Elgersma, David G. Amaral, Steven J. Gray, Benjamin D. Philpot
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Research Article Neuroscience Therapeutics

Dual-isoform hUBE3A gene transfer improves behavioral and seizure outcomes in Angelman syndrome model mice

Abstract

Loss of the maternal UBE3A allele causes Angelman syndrome (AS), a debilitating neurodevelopmental disorder. Here, we devised an AS treatment strategy based on reinstating dual-isoform expression of human UBE3A (hUBE3A) in the developing brain. Kozak sequence engineering of our codon-optimized vector (hUBE3Aopt) enabled translation of both short and long hUBE3A protein isoforms at a near-endogenous 3:1 (short/long) ratio, a feature that could help to support optimal therapeutic outcomes. To model widespread brain delivery and early postnatal onset of hUBE3A expression, we packaged the hUBE3Aopt vector into PHP.B capsids and performed intracerebroventricular injections in neonates. This treatment significantly improved motor learning and innate behaviors in AS mice, and it rendered them resilient to epileptogenesis and associated hippocampal neuropathologies induced by seizure kindling. hUBE3A overexpression occurred frequently in the hippocampus but was uncommon in the neocortex and other major brain structures; furthermore, it did not correlate with behavioral performance. Our results demonstrate the feasibility, tolerability, and therapeutic potential for dual-isoform hUBE3A gene transfer in the treatment of AS.

Authors

Matthew C. Judson, Charles Shyng, Jeremy M. Simon, Courtney R. Davis, A. Mattijs Punt, Mirabel T. Salmon, Noah W. Miller, Kimberly D. Ritola, Ype Elgersma, David G. Amaral, Steven J. Gray, Benjamin D. Philpot

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Figure 5

Neonatal ICV injection of PHP. B/hUBE3Aopt rescues motor learning and innate behaviors in AS mice.

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Neonatal ICV injection of PHP. B/hUBE3Aopt rescues motor learning and in...
(A) Experimental timeline for evaluation of behavioral performance in adult AS mice following neonatal ICV administration of 1 μL of 1.6 × 1014 vg/mL PHP.B/hUBE3Aopt. Sample sizes for each experimental group are listed to the right. (B) Distance traveled in the open field, per 5 minutes (left panel) and total (right panel). Two-way repeated-measures ANOVA and 1-way ANOVA, Tukey’s post hoc. (C) Latency to fall off the accelerating rotarod during the acquisition and retest phases of the task. Two-way repeated-measures ANOVA, Tukey’s post hoc. (D) Top row: representative images of marble burying arenas before (far left) and following 30-minute test sessions. Percentage of marbles buried, as determined by manual counting, is plotted to the right. Bottom row: representative thresholded images of marble burying arenas before (far left) and following 30-minute test sessions. Percentage of marble area obscured by bedding is plotted to the right. Welch’s 1-way ANOVA, Dunnett’s post hoc. (E) Representative images of nests built by mice of each treatment group with inset scores for nest quality. Middle panel: graph of nest building scores. Kruskal-Wallis test followed by Mann-Whitney U post hoc with Bonferroni correction for multiple comparisons. Right panel: mean ± SEM nesting material used during the 5-day nest building assay. Two-way repeated-measures ANOVA, Tukey’s post hoc. Data are represented as means ± SEM except for nest building scores. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.