Tight Long-term Dynamic Doxycycline Responsive Nigrostriatal GDNF Using a Single rAAV Vector
Tight Long-term Dynamic Doxycycline Responsive Nigrostriatal GDNF Using a Single rAAV Vector
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DOI：
10.1038/mt.2009.196
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发表时间：
2009-11-01
影响因子：
12.4
通讯作者：
Mandel, Ronald J.
中科院分区：
文献类型：
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作者：
Manfredsson, Fredric P.;Burger, Corinna;Mandel, Ronald J.
神经胶质细胞系来源的神经营养因子（GDNF）基因转移是帕金森病（PD）的一种治疗方法。由于潜在的副作用，外部转基因调控应该提高这种策略的安全性。在这里，我们使用基于重组腺相关病毒（rAAV）的双声四环素（tet）关闭构建物，证明了GDNF长期表达过程中的动态控制。黑质纹状体GDNF过表达可诱导啮齿动物体重改变，从而实现了黑质载体传递后GDNF在体内的纵向追踪。受调节的GDNF表达对膳食多西环素（DOX）高度敏感，当血清DOX水平低于抗菌活性所需水平时，纹状体GDNF水平无法检测到。然而，在缺乏DOX的情况下，纹状体gdnf水平超过了PD模型中有效所需的水平。我们还证明了当使用直接的细胞壁内载体递送时，没有一系列已知的与gdnf相关的副作用。因此，这种单一的rAAV载体系统满足了PD治疗实验试剂的大部分要求。
Glial cell line-derived neurotrophic factor (GDNF) gene transfer is being developed as a treatment for Parkinson's disease (PD). Due to the potential for side effects, external transgene regulation should enhance this strategy's safety profile. Here, we demonstrate dynamic control during long-term expression of GDNF using a recombinant adeno-associated virus (rAAV)-based bicistronic tetracycline (tet)-off construct. Nigrostriatal GDNF overexpression induces body weight alterations in rodents, enabling longitudinal in vivo tracking of GDNF expression after nigral vector delivery. Regulated GDNF-expression was highly sensitive to dietary doxycycline (DOX), displaying undetectable striatal GDNF levels at serum DOX levels below those required for antimicrobial activity. However, in the absence of DOX, striatal GDNF-levels exceeded levels required for efficacy in PD models. We also demonstrate the absence of a series of known GDNF-associated side effects when using direct intrastriatal vector delivery. Therefore, this single rAAV vector system meets most of the requirements for an experimental reagent for treatment of PD.