McCown T J, Xiao X, Li J, Breese G R, Samulski R J
Brain and Development Research Center, University of North Carolina at Chapel Hill 27599, USA.
Brain Res. 1996 Mar 25;713(1-2):99-107. doi: 10.1016/0006-8993(95)01488-8.
Safe, long-term gene expression is a primary criteria for effective gene therapy in the brain, so studies were initiated to evaluate adeno-associated virus (AAV) vector transfer of a reporter gene into specific sites of the rat brain. In the 4 day old rat, site infusions of AAV-CMV-lacZ (1 microliter; 5 x 10(4) particles) produced neuronal beta-galactosidase gene expression 3 weeks later in the hippocampus and inferior colliculus, but not in the cerebral cortex. Seven days after infusion of AAV-CMV-lacZ viral vectors (1 microliter) in the adult rat, beta-galactosidase gene expression was found in the olfactory tubercle, caudate, hippocampus, piriform cortex and inferior colliculus. primarily in multipolar neurons close to the infusion site. Three months after vector microinfusion, similar levels of gene expression remained in the olfactory tubercle and the inferior colliculus, with some reduction found in the caudate, but substantial reductions in beta-galactosidase gene expression occurred in the hippocampus and piriform cortex. In no case were obvious signs of toxicity noted. Therefore, AAV vectors can transfer foreign genes into the adult and neonatal CNS, but the pattern and longevity of gene expression depends upon the area of brain being studied.
安全、长期的基因表达是脑部有效基因治疗的主要标准，因此开展了相关研究来评估腺相关病毒（AAV）载体将报告基因转移至大鼠脑特定部位的情况。在4日龄大鼠中，向特定部位注射AAV-CMV-lacZ（1微升；5×10⁴个颗粒），3周后在海马体和下丘中产生了神经元β-半乳糖苷酶基因表达，但在大脑皮层中未产生。在成年大鼠中向特定部位注射AAV-CMV-lacZ病毒载体（1微升）7天后，在嗅结节、尾状核、海马体、梨状皮层和下丘中发现了β-半乳糖苷酶基因表达，主要在靠近注射部位的多极神经元中。载体微量注射3个月后，嗅结节和下丘中仍保持相似水平的基因表达，尾状核中的表达有所减少，而海马体和梨状皮层中的β-半乳糖苷酶基因表达则大幅减少。在任何情况下均未观察到明显的毒性迹象。因此，AAV载体可将外源基因转移至成年和新生中枢神经系统，但基因表达的模式和持续时间取决于所研究的脑区。
