鞘内注射 AAV9 后,物理定位显著增强了脑转导。
Physical positioning markedly enhances brain transduction after intrathecal AAV9 infusion.
机构信息
Department of Neurosciences, University of California, San Diego, La Jolla, CA 92093, USA.
Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
出版信息
Sci Adv. 2018 Nov 14;4(11):eaau9859. doi: 10.1126/sciadv.aau9859. eCollection 2018 Nov.
Several neurological disorders may benefit from gene therapy. However, even when using the lead vector candidate for intrathecal administration, adeno-associated virus serotype 9 (AAV9), the strength and distribution of gene transfer to the brain are inconsistent. On the basis of preliminary observations that standard intrathecal AAV9 infusions predominantly drive reporter gene expression in brain regions where gravity might cause cerebrospinal fluid to settle, we tested the hypothesis that counteracting vector "settling" through animal positioning would enhance vector delivery to the brain. When rats are either inverted in the Trendelenburg position or continuously rotated after intrathecal AAV9 infusion, we find (i) a significant 15-fold increase in the number of transduced neurons, (ii) a marked increase in gene delivery to cortical regions, and (iii) superior animal-to-animal consistency of gene expression. Entorhinal, prefrontal, frontal, parietal, hippocampal, limbic, and basal forebrain neurons are extensively transduced: 95% of transduced cells are neurons, and greater than 70% are excitatory. These findings provide a novel and simple method for broad gene delivery to the cortex and are of substantial relevance to translational programs for neurological disorders, including Alzheimer's disease and related dementias, stroke, and traumatic brain injury.
几种神经紊乱疾病可能受益于基因治疗。然而,即使使用鞘内给药的主要载体候选物腺相关病毒血清型 9(AAV9),基因向大脑的转移的强度和分布也不一致。基于初步观察到标准鞘内 AAV9 输注主要在重力可能导致脑脊液沉降的脑区驱动报告基因表达,我们测试了通过动物定位来对抗载体“沉降”会增强载体向大脑的传递的假设。当大鼠在特伦德伦伯体位或在鞘内 AAV9 输注后连续旋转时,我们发现:(i)转导神经元的数量显著增加了 15 倍,(ii)向皮质区的基因传递明显增加,以及(iii)基因表达的动物间一致性更好。内嗅、前额叶、额叶、顶叶、海马、边缘和基底前脑神经元被广泛转导:95%的转导细胞为神经元,超过 70%为兴奋性神经元。这些发现为广泛向皮质传递基因提供了一种新颖而简单的方法,对于神经紊乱疾病的转化计划,包括阿尔茨海默病和相关痴呆症、中风和创伤性脑损伤具有重要意义。
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