Department of Neurophysiology and Neural Repair, Gunma University Graduate School of Medicine, 3-39-22, Gunma, 371-8511, Japan; Department of Biomedical Science, Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia.
Department of Neurophysiology and Neural Repair, Gunma University Graduate School of Medicine, 3-39-22, Gunma, 371-8511, Japan; Viral Vector Core, Gunma University Initiative for Advanced Research (GIAR), Gunma, 371-8511, Japan.
Neurosci Lett. 2021 Jun 21;756:135956. doi: 10.1016/j.neulet.2021.135956. Epub 2021 May 11.
Adeno-associated virus (AAV)- PHP.B and AAV-PHP.eB (PHP.eB), a capsid variant of AAV serotype 9, efficiently penetrates the mouse blood-brain barrier and predominantly infects neurons. Thus, the PHP.B / PHP.eB capsid and a neuron-specific promoter is a reasonable combination for effective neuronal transduction. However, the transduction characteristics of intravenously administered PHP.B / PHP.eB carrying different neuron-specific promoters have not been studied systematically. In this study, using an intravenous infusion of PHP.eB in mice, we performed a comparative study of the ubiquitous CBh and three neuron-specific promoters, the Ca/calmodulin-dependent kinase subunit α (CaMKII) promoter, neuron-specific enolase (NSE) promoter, and synapsin I with a minimal CMV sequence (SynI-minCMV) promoter. Expression levels of a transgene by three neuron-specific promoters were comparable to or higher than those of the CBh promoter. Among the promoters examined, the NSE promoter showed the highest transgene expression. All neuron-specific promoters were activated specifically in the neurons. PHP.eB carrying the CaMKII promoter, which is generally believed to exert its function exclusively in the excitatory neurons, transduced both the excitatory and inhibitory neurons without bias, whereas PHP.eB with the NSE and SynI-minCMV promoters transduced neurons with significant bias toward inhibitory neurons. These results are useful in neuron-targeted broad transgene expression through systemic infusion of blood-brain-barrier-penetrating AAV vectors carrying the neuron-specific promoter.
腺相关病毒（AAV）- PHP.B 和 AAV-PHP.eB（PHP.eB），是血清型 9 的衣壳变体，能够有效地穿透小鼠血脑屏障并主要感染神经元。因此，PHP.B/PHP.eB 衣壳和神经元特异性启动子是实现有效神经元转导的合理组合。然而，静脉内给予携带不同神经元特异性启动子的 PHP.B/PHP.eB 的转导特性尚未被系统地研究过。在这项研究中，我们使用静脉内输注 PHP.eB 在小鼠中进行了一项比较研究，研究了广泛表达的 CBh 和三个神经元特异性启动子，即钙/钙调蛋白依赖性激酶亚单位α（CaMKII）启动子、神经元特异性烯醇化酶（NSE）启动子和最小 CMV 序列（SynI-minCMV）启动子。三个神经元特异性启动子的转基因表达水平与 CBh 启动子相当或更高。在研究的启动子中，NSE 启动子显示出最高的转基因表达。所有神经元特异性启动子都特异性地在神经元中被激活。携带 CaMKII 启动子的 PHP.eB 通常被认为仅在兴奋性神经元中发挥作用，但它没有偏向地转导兴奋性和抑制性神经元，而携带 NSE 和 SynI-minCMV 启动子的 PHP.eB 则明显偏向于转导抑制性神经元。这些结果对于通过携带神经元特异性启动子的血脑屏障穿透性 AAV 载体的全身输注实现针对神经元的广泛转基因表达是有用的。
