Cearley Cassia N, Wolfe John H
Walter Flato Goodman Center for Comparative Medical Genetics, School of Veterinary Medicine, University of Pennsylvania, and Division of Neurology, Stokes Research Institute, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA.
J Neurosci. 2007 Sep 12;27(37):9928-40. doi: 10.1523/JNEUROSCI.2185-07.2007.
Neurogenetic disorders typically affect cells throughout the brain. Adeno-associated virus (AAV) vector-mediated transfer of a normal cDNA can correct the metabolic defects at the site of injection, but treatment of the entire brain requires widespread delivery of the normal gene and/or protein. Current methods require multiple injections for widespread distribution. However, some AAV vectors can be transported along neuronal pathways associated with the injected region. Thus, targeting widely dispersed systems in the CNS might be a pathway for gene dispersal from a limited number of sites. We tested this hypothesis in the ventral tegmental area (VTA), a region with numerous efferent and afferent projections. A single 1 mul injection resulted in transport of the vector genome to projection sites in distal parts of the brain. When compared with injections into the striatum, the VTA injection resulted in higher enzyme levels in more regions of the brain. The AAV-9 serotype vector was the most widely disseminated, but AAV-Rh.10 and AAV-1 were also transported after VTA injection. The effect on global lesions of a neurogenetic disease was tested in the mouse model of MPS VII (mucopolysaccharidosis VII), a lysosomal storage disorder. Widespread distribution of the vector genome after AAV-9 VTA injection resulted in even further distribution of the enzyme product, by secretion and uptake by surrounding cells, and complete correction of the storage lesions throughout the entire brain. This unprecedented level of correction from a single injection into the developed brain provides a potential strategy to correct a large volume of brain while minimizing the number of injections.
神经遗传性疾病通常会影响整个大脑的细胞。腺相关病毒(AAV)载体介导的正常互补DNA的转移可以纠正注射部位的代谢缺陷,但治疗整个大脑需要广泛递送正常基因和/或蛋白质。目前的方法需要多次注射才能广泛分布。然而,一些AAV载体可以沿着与注射区域相关的神经通路运输。因此,靶向中枢神经系统中广泛分散的系统可能是从有限数量的位点进行基因扩散的途径。我们在腹侧被盖区(VTA)测试了这一假设,该区域有许多传出和传入投射。单次1微升注射导致载体基因组运输到大脑远端的投射部位。与注射到纹状体相比,VTA注射在大脑更多区域导致更高的酶水平。AAV-9血清型载体传播最广,但VTA注射后AAV-Rh.10和AAV-1也被运输。在黏多糖贮积症VII(MPS VII)的小鼠模型中测试了对神经遗传性疾病整体病变的影响,MPS VII是一种溶酶体贮积症。AAV-9 VTA注射后载体基因组的广泛分布导致酶产物通过周围细胞的分泌和摄取进一步分布,并完全纠正了整个大脑的贮积性病变。这种对发育成熟大脑单次注射前所未有的纠正水平提供了一种潜在策略,可在尽量减少注射次数的同时纠正大量脑组织。