Improved method of recombinant AAV2 delivery for systemic targeted gene therapy
Improved method of recombinant AAV2 delivery for systemic targeted gene therapy
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DOI：
10.1006/mthe.2001.0636
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发表时间：
2002-07-01
影响因子：
12.4
通讯作者：
Byrne, BJ
中科院分区：
文献类型：
--
作者：
Mah, C;Fraites, TJ;Byrne, BJ
目前大多数基因治疗方式的主要障碍是以非常高的效率转染靶组织的能力，这最终导致治疗水平的转基因表达。我们已经开发了一种新的方法，重组腺相关病毒2（rAAV）的交付，结果在增加载体转导效率使用可逆共轭微球rAAV载体。我们假设与微球的缀合应导致载体的更高有效浓度以及载体在其移动通过组织脉管系统时对靶细胞的更长相对暴露时间。体外实验表明，当与微球缀合时，使用1%的载体可以实现与游离载体相同的转导水平。此外，使用磁性微球，可以靶向感染区域。在体内，我们证明了微球介导的rAAV载体的递送在肌内或静脉内施用时比单独使用游离载体的递送导致更高的转导效率。此外，我们证明了通过在毛细血管床中保留微球结合的载体来将转基因表达靶向至特定组织。这些研究证明了一种更有效地递送rAAV载体的新方法，可以证明是病毒介导的人类基因治疗的成功替代模式。
A major hurdle in most current gene therapy modalities is the ability to transduce target tissues at very high efficiencies that ultimately lead to therapeutic levels of transgene expression. We have developed a novel method of recombinant adeno-associated virus 2 (rAAV) delivery that results in increased vector transduction efficiencies using microspheres reversibly conjugated to rAAV vectors. We hypothesize that conjugation to microspheres should result in a higher effective concentration of vector as well as longer relative exposure time of vector to target cells as it moves through the tissue vasculature. In vitro experiments demonstrate that the same level of transduction seen with free vector can be achieved using 1% of vector when conjugated to microspheres. In addition, using magnetic microspheres, the region of infection can be targeted. In vivo, we demonstrate that microsphere-mediated delivery of rAAV vector results in higher transduction efficiencies than delivery with free vector alone when administered either intramuscularly or intravenously. Furthermore, we demonstrate targeting of transgene expression to specific tissues by retention of microsphere-bound vector in the capillary bed. These studies demonstrate a novel method to deliver rAAV vectors more effectively that could prove to be a successful alternative mode of virus-mediated human gene therapy.