Self-inactivating lentiviral vectors with enhanced transgene expression as potential gene transfer system in Parkinson's disease
Self-inactivating lentiviral vectors with enhanced transgene expression as potential gene transfer system in Parkinson's disease
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DOI：
10.1089/10430340050016256
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发表时间：
2000-01-01
影响因子：
4.2
通讯作者：
Aebischer, P
中科院分区：
文献类型：
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作者：
Déglon, N;Tseng, JL;Aebischer, P
胶质细胞源性神经营养因子（GDNF）能够保护多巴胺能神经元免受各种损伤，因此构成治疗帕金森病的有希望的候选者。感染静止神经元细胞的慢病毒载体可以允许GDNF的局部递送，从而避免与其他脑结构的激活相关的潜在副作用，为了在确保最大生物安全性和最佳转基因表达的情况下测试该假设，通过插入土拨鼠肝炎病毒的转录后调节元件来修饰自失活（SIN）慢病毒载体，并且用多重衰减包装系统生产颗粒。在成年大鼠的黑质中单次注射2 μ l lacZ表达载体（SIN-W-LacZ）后，平均40.1 +/-6.0%的酪氨酸羟化酶（TH）阳性神经元被转导，而第一代慢病毒载体为5.0 +/-2.1%。此外，在小鼠磷酸甘油酸激酶1（PGK）启动子控制下表达GDNF的SIN-W载体能够保护内侧前脑束轴突切断后的黑质多巴胺能神经元。在注射SLN-W-PGK-GDNF载体的动物的中脑提取物中检测到纳克范围的hGDNF表达，而在注射对照载体的动物中检测不到。具有增强的表达和安全特征的慢病毒载体进一步确立了这些载体用于将生物活性分子局部递送到中枢神经系统的限定结构中的潜在用途。
Glial cell line-derived neurotrophic factor (GDNF) is able to protect dopaminergic neurons against various insults and constitutes therefore a promising candidate for the treatment of Parkinson's disease, Lentiviral vectors that infect quiescent neuronal cells may allow the localized delivery of GDNF, thus avoiding potential side effects related to the activation of other brain structures, To test this hypothesis in a setting ensuring both maximal biosafety and optimal transgene expression, a self-inactivating (SIN) lentiviral vector was modified by insertion of the posttranscriptional regulatory element of the woodchuck hepatitis virus, and particles were produced with a multiply attenuated packaging system. After a single injection of 2 mu l of a lacZ-expressing vector (SIN-W-LacZ) in the substantia nigra of adult rats, an average of 40.1 +/- 6.0% of the tyrosine hydroxylase (TH)-positive neurons were transduced as compared with 5.0 +/- 2.1% with the first-generation lentiviral vector. Moreover, the SIN-W vector expressing GDNF under the control of the mouse phosphoglycerate kinase 1 (PGK) promoter was able to protect nigral dopaminergic neurons after medial forebrain bundle axotomy. Expression of hGDNF in the nanogram range vas detected in extracts of mesencephalon of animals injected with an SLN-W-PGK-GDNF vector, whereas it was undetectable in animals injected with a control vector, Lentiviral vectors with enhanced expression and safety features further establish the potential use of these vectors for the local delivery of bioactive molecules into defined structures of the central nervous system.