Department of Medical Biology and Genetics, Kazan State Medical University, 420012 Kazan, Russia.
Institute of Fundamental Medicine and Biology, Kazan [Volga Region] Federal University, 420008 Kazan, Russia.
Int J Mol Sci. 2020 Sep 18;21(18):6858. doi: 10.3390/ijms21186858.
Currently, the main fundamental and clinical interest for stroke therapy is focused on developing a neuroprotective treatment of a penumbra region within the therapeutic window. The development of treatments for ischemic stroke in at-risk patients is of particular interest. Preventive gene therapy may significantly reduce the negative consequences of ischemia-induced brain injury. In the present study, we suggest the approach of preventive gene therapy for stroke. Adenoviral vectors carrying genes encoding vascular endothelial growth factor (VEGF), glial cell-derived neurotrophic factor (GDNF) and neural cell adhesion molecule (NCAM) or gene engineered umbilical cord blood mononuclear cells (UCB-MC) overexpressing recombinant VEGF, GDNF, and NCAM were intrathecally injected before distal occlusion of the middle cerebral artery in rats. Post-ischemic brain recovery was investigated 21 days after stroke modelling. Morphometric and immunofluorescent analysis revealed a reduction of infarction volume accompanied with a lower number of apoptotic cells and decreased expression of Hsp70 in the peri-infarct region in gene-treated animals. The lower immunopositive areas for astrocytes and microglial cells markers, higher number of oligodendrocytes and increased expression of synaptic proteins suggest the inhibition of astrogliosis, supporting the corresponding myelination and functional recovery of neurons in animals receiving preventive gene therapy. In this study, for the first time, we provide evidence of the beneficial effects of preventive triple gene therapy by an adenoviral- or UCB-MC-mediated intrathecal simultaneous delivery combination of and on the preservation and recovery of the brain in rats with subsequent modelling of stroke.
目前，针对中风治疗的主要基础和临床兴趣集中在开发治疗半影区的神经保护治疗上。因此，开发有中风风险的患者的治疗方法尤其具有意义。预防性基因治疗可能会显著降低缺血性脑损伤的负面后果。在本研究中，我们提出了预防性基因治疗中风的方法。携带血管内皮生长因子（VEGF）、胶质细胞源性神经营养因子（GDNF）和神经细胞黏附分子（NCAM）编码基因的腺病毒载体或过表达重组 VEGF、GDNF 和 NCAM 的基因工程脐带血单核细胞（UCB-MC）被鞘内注射到大鼠大脑中动脉远端闭塞之前。在中风模型建立 21 天后，研究了缺血后大脑的恢复情况。形态计量学和免疫荧光分析显示，基因治疗动物的梗死体积减少，同时梗死周围区域的凋亡细胞数量减少，Hsp70 表达降低。少突胶质细胞和星形胶质细胞标志物的免疫阳性区域减少，少突胶质细胞数量增加，突触蛋白表达增加，表明接受预防性基因治疗的动物的星形胶质细胞增生受到抑制，支持神经元的相应髓鞘形成和功能恢复。在这项研究中，我们首次提供了通过腺病毒或 UCB-MC 介导的鞘内同时传递 和 的组合进行预防性三重基因治疗对中风后大鼠大脑保存和恢复的有益效果的证据。
