Effects of gamma irradiation on the transduction of dividing and nondividing cells in brain and muscle of rats by adeno-associated virus vectors
Effects of gamma irradiation on the transduction of dividing and nondividing cells in brain and muscle of rats by adeno-associated virus vectors
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DOI：
10.1089/hum.1996.7.7-841
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发表时间：
1996-05-01
影响因子：
4.2
通讯作者：
Miller, AD
中科院分区：
文献类型：
--
作者：
Alexander, IE;Russell, DW;Miller, AD
基于腺相关病毒（AAV）的载体正在研究用于基因治疗的应用。AAV载体生物学的关键方面仍未确定，特别是介导转导和决定宿主细胞转导许可的细胞内事件和活动。使用培养的原代人成纤维细胞，我们之前发现AAV载体优先（但不是唯一）转导细胞周期的S期，并且通过干扰DNA代谢的物理和化学试剂对靶细胞进行预处理可以显著增强转导。在这项研究中，我们测试了AAV载体性能的类似改善是否可以在体内实现。选择成年大鼠的大脑和覆盖的头皮肌肉作为载体接种，因为存在定义明确的分裂、静止和有丝分裂后细胞群体，并且选择伽马辐射作为诱导这些细胞DNA修复的可复制手段。我们发现，伽马辐射显著增强了中枢神经系统中蛛网膜上皮和脉络膜上皮以及头皮中成熟的非分裂肌肉细胞的分裂细胞群的转导，而伽马辐射并没有增加海马有丝分裂后神经元的基础转导水平。这些数据证实，AAV载体的转导不需要复制性细胞DNA合成，并表明靶细胞的有丝分裂状态不一定能预测对转导增强治疗的反应。最重要的是，这些数据表明，可以在体内操纵靶细胞，使其更容易接受AAV载体转导。
Vectors based on adeno-associated virus (AAV) are under investigation for use in gene therapy applications. Critical aspects of AAV vector biology remain undefined, in particular the intracellular events and activities mediating transduction and determining host cell permissiveness for transduction. Using cultured primary human fibroblasts, we previously showed that AAV vectors preferentially, but not exclusively, transduce cells in the S phase of the cell cycle, and that transduction can be markedly enhanced by pretreatment of target cells with physical and chemical agents that perturb DNA metabolism. In this study, we tested whether similar improvements in AAV vector performance might be achievable in vivo. The adult rat brain and overlying scalp muscle mere selected for vector inoculation because of the presence of well-defined populations of dividing, quiescent, and post-mitotic cells, and gamma irradiation was chosen as a reproducible means of inducing DNA repair in these cells. We find that gamma irradiation markedly enhances the transduction of dividing cell populations in the pia-arachnoid and choroid epithelium within the central nervous system, and of mature nondividing muscle cells in the scalp, whereas gamma irradiation did not increase the basal transduction level of post-mitotic neurons in the hippocampus. These data confirm that replicative cellular DNA synthesis is not required for transduction by AAV vectors and show that the mitotic state of target cells is not necessarily predictive of responsiveness to transduction-enhancing treatments. Most importantly, these data demonstrate that target cells can be manipulated in vivo to render them more permissive for AAV vector transduction.