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Abstract：
Lafora disease is a rare and fatal form of progressive myoclonic epilepsy typically occurring early in adolescence. The disease results from mutations in the EPM2A gene, encoding laforin, or the EPM2B gene, encoding malin. Laforin and malin work together in a complex to control glycogen synthesis and prevent the toxicity produced by misfolded proteins via the ubiquitin-proteasome system. Disruptions in either protein cause alterations in this complex, leading to the formation of Lafora bodies containing abnormal, insoluble, and hyperphosphorylated forms of glycogen. We used the Epm2a-/- knockout mouse model of Lafora disease to apply gene therapy by administering intracerebroventricular injections of a recombinant adeno-associated virus carrying the human EPM2A gene. We evaluated the effects of this treatment through neuropathological studies, behavioral tests, video-electroencephalography, electrophysiological recordings, and proteomic/phosphoproteomic analysis. Gene therapy ameliorated neurological and histopathological alterations, reduced epileptic activity and neuronal hyperexcitability, and decreased the formation of Lafora bodies. Moreover, differential quantitative proteomics and phosphoproteomics revealed beneficial changes in various molecular pathways altered in Lafora disease. Our results represent proof of principle for gene therapy with the coding region of the human EPM2A gene as a treatment for EPM2A-related Lafora disease.
摘要：
Lafora病是一种罕见且致命的进行性肌阵挛性癫痫，通常发生在青春期早期。这种疾病是由EPM2A基因突变引起的，编码拉福林，或者EPM2B基因，编码Malin.Laforin和malin在复合物中一起工作以控制糖原合成并防止错误折叠的蛋白质通过泛素-蛋白酶体系统产生的毒性。任何一种蛋白质的破坏都会导致这种复合物的改变，导致形成含有异常的拉福拉体，不溶性,和糖原的过度磷酸化形式。我们使用Lafora病的Epm2a-/-敲除小鼠模型通过侧脑室注射携带人EPM2A基因的重组腺相关病毒来应用基因治疗。我们通过神经病理学研究评估了这种治疗的效果，行为测试,视频脑电图,电生理记录，和蛋白质组/磷酸化蛋白质组分析。基因治疗改善了神经和组织病理学改变，减少癫痫活动和神经元过度兴奋，并减少了Lafora身体的形成。此外，差异定量蛋白质组学和磷酸化蛋白质组学揭示了Lafora疾病中各种分子途径改变的有益变化。我们的结果代表了人EPM2A基因编码区的基因治疗作为EPM2A相关Lafora疾病治疗的原理证明。
