Emergency Department, Department of Neurology of the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
Department of Neurology of the Sixth Affiliated Hospital of Guangzhou Medical University, Department of Human Anatomy in School of Basic Science of Guangzhou Medical University, Key Laboratory of Neurogenetics and Channelopathies of Guangdong Province and the Ministry of Education of China, Guangzhou, China.
Aging Dis. 2023 Dec 20;15(6):2632-2649. doi: 10.14336/AD.2023.1213.
AAV-PHP.eB depends on endothelial cells to highly transduce the central nervous system (CNS) and is widely used for intravenous gene therapy. However, the transduction profile and therapeutic efficiency after endothelial cell injury such as ischemic stroke is largely unknown. In this study, we tested the transduction profiles of AAV-PHP.eB and developed intravenous NeuroD1 gene therapy to treat ischemic stroke in mice. We found that AAV-PHP.eB-GFP control virus crossed the BBB and infected brain cells efficiently in normal brain. However, after stroke, AAV-PHP.eB-GFP control virus was highly restricted in the blood vessels. Surprisingly, after switching to therapeutic vector AAV-PHP.eB-NeuroD1-GFP, the viral vector successfully crossed blood vessels and infected brain cells. Using Tie2-cre transgenic mice, we demonstrated that NeuroD1 regulated endothelial gene expression to modulate AAV-PHP.eB transduction. Following the changes of signaling pathways in endothelial cells, NeuroD1 effectively protected BBB integrity, attenuated neuroinflammation, inhibited neuron apoptosis and rescued motor deficits after ischemic stroke. Moreover, NeuroD1 over-expression in brain cells further promoted neural regeneration. These results indicate that intravenous gene therapy using AAV-PHP.eB for ischemic stroke differs from intracranial gene therapy and NeuroD1 intravenous delivery using AAV-PHP.eB efficiently rescue both vascular damage and neuronal loss, providing an advancing therapeutic treatment for stroke.
AAV-PHP.eB 依赖于血管内皮细胞来高效转导中枢神经系统 (CNS)，广泛用于静脉内基因治疗。然而，在缺血性卒中等血管内皮细胞损伤后，AAV-PHP.eB 的转导谱和治疗效率在很大程度上仍不清楚。在这项研究中，我们测试了 AAV-PHP.eB 的转导谱，并开发了静脉内 NeuroD1 基因治疗来治疗小鼠的缺血性卒中。我们发现 AAV-PHP.eB-GFP 对照病毒在正常大脑中能够穿过血脑屏障并有效地感染脑细胞。然而，在卒中后，AAV-PHP.eB-GFP 对照病毒在血管中受到高度限制。令人惊讶的是，在切换到治疗性载体 AAV-PHP.eB-NeuroD1-GFP 后，病毒载体成功地穿过血管并感染了脑细胞。利用 Tie2-cre 转基因小鼠，我们证明了 NeuroD1 通过调节 AAV-PHP.eB 的转导来调节血管内皮细胞的基因表达。随着内皮细胞信号通路的变化，NeuroD1 有效地保护了 BBB 的完整性，减轻了神经炎症，抑制了卒中后的神经元凋亡，并挽救了运动功能障碍。此外，NeuroD1 在脑细胞中的过表达进一步促进了神经再生。这些结果表明，AAV-PHP.eB 用于缺血性卒中的静脉内基因治疗与颅内基因治疗不同，AAV-PHP.eB 静脉内递送 NeuroD1 可有效挽救血管损伤和神经元丢失，为卒中提供了一种先进的治疗方法。
