Transgene Expression in Dogs After Liver-Directed Hydrodynamic Delivery of Sleeping Beauty Transposons Using Balloon Catheters
Transgene Expression in Dogs After Liver-Directed Hydrodynamic Delivery of Sleeping Beauty Transposons Using Balloon Catheters
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DOI：
10.1089/hum.2017.003
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发表时间：
2017-07-01
影响因子：
4.2
通讯作者：
McIvor, R. Scott
中科院分区：
文献类型：
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作者：
Hyland, Kendra A.;Aronovich, Elena L.;McIvor, R. Scott
睡美人转座子系统已被广泛测试用于在小鼠体内和体外整合报告基因和治疗基因。狗被用作人类治疗和DNA溶液微创输注的大型动物模型。使用球囊导管将DNA溶液递送到整个肝脏或肝脏左侧，用于暂时阻塞静脉流出。在80和140毫米汞柱之间的峰值血管内压力支持足够的DNA输送在狗的肝脏检测分泌报告蛋白。可分泌的报告者允许监测输注后循环中可检测到的基因产物的时间过程。在血浆中测量犬分泌的碱性磷酸酶报告蛋白水平，表达可检测长达6周，而犬促红细胞生成素的表达可检测7-10天。所有动物的血液转氨酶均出现一过性升高，并在10天内恢复正常;除此之外，给药动物的临床表现正常。这些结果证明了分泌的报告蛋白在大型动物模型中用于实时监测肝脏中基因表达的实用性，但强调了需要改善靶组织中的递送以支持睡美人转座子的整合和长期表达。
The Sleeping Beauty transposon system has been extensively tested for integration of reporter and therapeutic genes in vitro and in vivo in mice. Dogs were used as a large animal model for human therapy and minimally invasive infusion of DNA solutions. DNA solutions were delivered into the entire liver or the left side of the liver using balloon catheters for temporary occlusion of venous outflow. A peak intravascular pressure between 80 and 140 mmHg supported sufficient DNA delivery in dog liver for detection of secretable reporter proteins. Secretable reporters allowed monitoring of the time course of gene products detectable in the circulation postinfusion. Canine secreted alkaline phosphatase reporter protein levels were measured in plasma, with expression detectable for up to 6 weeks, while expression of canine erythropoietin was detectable for 7-10 days. All animals exhibited a transient increase in blood transaminases that normalized within 10 days; otherwise the treated animals were clinically normal. These results demonstrate the utility of a secreted reporter protein for real-time monitoring of gene expression in the liver in a large animal model but highlight the need for improved delivery in target tissues to support integration and long-term expression of Sleeping Beauty transposons.