Long-term correction of ornithine Transcarbamylase deficiency by WPRE-mediated overexpression using a helper-dependent adenovirus
Long-term correction of ornithine Transcarbamylase deficiency by WPRE-mediated overexpression using a helper-dependent adenovirus
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DOI：
10.1016/j.ymthe.2004.05.036
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发表时间：
2004-09-01
影响因子：
12.4
通讯作者：
Lee, B
中科院分区：
文献类型：
--
作者：
Mian, A;McCormack, M;Lee, B
尿素循环障碍（UCD）是发展肝脏疾病基因替代治疗的重要模型。最常见的UCD，鸟氨酸转氨甲酰酶（OTC）缺乏症的长期纠正，尚未在临床或临床前环境中实现。使用早代腺病毒（Ad）的单一人体临床试验未能显示任何生化校正。在成年OTC缺陷型小鼠中，表达小鼠OTC基因但不表达人OTC基因的El/E2缺失的Ad载体仅具有瞬时治疗性。通过在免疫原性较低的辅助病毒依赖性腺病毒载体的背景下使用转录后过表达，我们实现了对成年OTC缺陷型小鼠的代谢校正>6个月。在无慢性肝毒性的情况下，乳清酸尿正常化，肝酶活性正常，OTC RNA和蛋白水平升高，证明了这一结果。过度表达的人类蛋白质可能克服了两个潜在的机制，占穷人的跨物种互补：在线粒体进口水平的动力学块或突变多肽的显性负效应。这些数据代表了治疗人类先天性肝细胞代谢缺陷的重要方法，如需要高水平transluction和基因表达进行临床纠正的UCD。
The urea cycle disorders (UCDs) are important models for developing gene replacement therapy for liver diseases. Long-term correction of the most common UCD, ornithine transcarbamylase (OTC) deficiency, has yet to be achieved in clinical or preclinical settings. The single human clinical trial using early-generation adenovirus (Ad) failed to show any biochemical correction. In adult OTC-deficient mice, an El/E2-deleted Ad vector expressing the mouse OTC gene, but not the human, was only transiently therapeutic. By using post-transcriptional overexpression in the context of the less immunogenic helper-dependent adenoviral vector, we achieved metabolic correction of adult OTC-deficient mice for >6 months. Demonstrating this result were normalized orotic aciduria, normal hepatic enzyme activity, and elevated OTC RNA and protein levels in the absence of chronic hepatotoxicity. Overexpressing the human protein may have overcome two potential mechanisms accounting for poor cross-species complementation: a kinetic block at the level of mitochondrial import or a dominant negative effect by the mutant polypeptide. These data represent an important approach for treating human inborn errors of hepatocyte metabolism like the UCDs that require high-level transcluction and gene expression for clinical correction.