Neuroscience Center.
Department of Cell Biology and Physiology.
JCI Insight. 2021 Oct 22;6(20):e144712. doi: 10.1172/jci.insight.144712.
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.
UBE3A 母源等位基因缺失会导致 Angelman 综合征（AS），这是一种使人衰弱的神经发育障碍。在这里，我们设计了一种基于在发育中的大脑中重新表达人类 UBE3A（hUBE3A）的双异构体的 AS 治疗策略。我们的密码子优化载体（hUBE3Aopt）的 Kozak 序列工程使短和长 hUBE3A 蛋白异构体以接近内源性的 3:1（短/长）比例进行翻译，这一特性有助于支持最佳的治疗效果。为了模拟 hUBE3A 在大脑中的广泛传递和出生后早期表达，我们将 hUBE3Aopt 载体包装到 PHP.B 衣壳中，并在新生鼠中进行侧脑室注射。这种治疗方法显著改善了 AS 小鼠的运动学习和先天行为，并使它们能够抵抗由癫痫发作引起的癫痫发生和相关的海马神经病理学。hUBE3A 过表达在海马体中频繁发生，但在新皮层和其他主要大脑结构中不常见；此外，它与行为表现无关。我们的结果表明，双异构体 hUBE3A 基因转移在治疗 AS 方面具有可行性、耐受性和治疗潜力。
