scAAV9 Intracisternal Delivery Results in Efficient Gene Transfer to the Central Nervous System of a Feline Model of Motor Neuron Disease
scAAV9 Intracisternal Delivery Results in Efficient Gene Transfer to the Central Nervous System of a Feline Model of Motor Neuron Disease
复制标题
DOI：
10.1089/hum.2012.218
复制
发表时间：
2013-07-01
影响因子：
4.2
通讯作者：
Joussemet, Beatrice
中科院分区：
文献类型：
--
作者：
Bucher, Thomas;Colle, Marie-Anne;Joussemet, Beatrice
在以前的研究表明，血管内皮生长因子（VEGF）可以保护运动神经元的退化，腺相关病毒载体（血清型1和9）编码的VEGF（AAV.VEGF）的肢体表达1（LIX1）缺陷的猫，下运动神经元疾病的大型动物模型，使用三种不同的传递途径，以中枢神经系统。将AAV.vegf载体通过脑内施用注射到年轻成年猫的运动皮质中、小脑延髓池中或静脉内注射。脑内注射导致可检测的转基因DNA和整个脊髓的转录本，确认通过皮质脊髓途径的AAV的顺行运输。然而，这种策略导致脊髓中VEGF表达水平较低。静脉内注射的类似AAV剂量也导致脊髓转导不良。相反，脑池内递送AAV在整个脊髓中表现出长期转导和高水平的VEGF表达，但在LIX1缺陷动物中没有可检测的治疗临床益处。总之，我们证明了（i）脑池内递送是一种有效的AAV递送途径，导致整个脊髓的高转导，与很少或没有脱靶基因表达相关，以及（ii）然而，在LIX1缺陷的猫模型中，脊髓中高水平表达的VEGF对疾病进程没有有益的影响。
On the basis of previous studies suggesting that vascular endothelial growth factor (VEGF) could protect motor neurons from degeneration, adeno-associated virus vectors (serotypes 1 and 9) encoding VEGF (AAV.vegf) were administered in a limb-expression 1 (LIX1)-deficient cat-a large animal model of lower motor neuron disease-using three different delivery routes to the central nervous system. AAV.vegf vectors were injected into the motor cortex via intracerebral administration, into the cisterna magna, or intravenously in young adult cats. Intracerebral injections resulted in detectable transgene DNA and transcripts throughout the spinal cord, confirming anterograde transport of AAV via the corticospinal pathway. However, such strategy led to low levels of VEGF expression in the spinal cord. Similar AAV doses injected intravenously resulted also in poor spinal cord transduction. In contrast, intracisternal delivery of AAV exhibited long-term transduction and high levels of VEGF expression in the entire spinal cord, yet with no detectable therapeutic clinical benefit in LIX1-deficient animals. Altogether, we demonstrate (i) that intracisternal delivery is an effective AAV delivery route resulting in high transduction of the entire spinal cord, associated with little to no off-target gene expression, and (ii) that in a LIX1-deficient cat model, however, VEGF expressed at high levels in the spinal cord has no beneficial impact on the disease course.