Gene Therapy And Neuroprotection

基因治疗和神经保护

• 批准号: 6830662
• 负责人: Yun Wang
• 金额: --
• 依托单位: NATIONAL INSTITUTE ON DRUG ABUSE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6830662
• 关键词: Alphaherpesvirinae; abnormal involuntary movement; artery occlusion; cerebral artery; cerebral ischemia /hypoxia; cysteine endopeptidases; enzyme activity; gene expression; gene therapy; glial fibrillary acidic protein; laboratory rat; neuroprotectants; neurotrophic factors; nonhuman therapy evaluation; psychomotor disorders; stroke; stroke therapy; transfection /expression vector

Direct intracerebral administration of glial cell line-derived neurotrophic factor (GDNF) is neuroprotective against ischemia-induced cerebral injury. Utilizing viral vectors to deliver and express therapeutic genes presents an opportunity to produce GDNF within localized regions of an evolving infarct. We investigated whether a Herpes simplex virus (HSV) amplicon-based vector encoding GDNF (HSVgdnf) would protect neurons against ischemic injury. In primary neuronal cultures HSVgdnf reduced oxidant?induced injury compared to the control vector HSVlac. To test protective effects in vivo, HSVgdnf or HSVlac was injected into the cerebral cortex prior to or subsequent to a 60 minute unilateral occlusion of the middle cerebral artery (MCA). Control stroke animals developed bradykinesia and motor asymmetry; pretreatment with HSVgdnf significantly reduced such motor deficits. Animals receiving HSVlac or HSVgdnf after the ischemic insult did not exhibit any behavioral improvement. Histological analyses performed one month after stroke revealed a reduction in ischemic tissue loss in rats pretreated with HSVgdnf. Similarly, these animals exhibited less immunostaining for glial fibrillary acidic protein (GFAP) and caspase-3. Taken together, our data indicate that HSVgdnf pretreatment provides protection against cerebral ischemia and supports the utilization of the HSV amplicon for therapeutic delivery of trophic factors to the CNS.

脑内直接注射胶质细胞源性神经营养因子(GDNF)对脑缺血损伤具有神经保护作用。利用病毒载体传递和表达治疗性基因，为在演变中的梗塞局部区域产生GDNF提供了机会。我们研究了单纯疱疹病毒(HSV)扩增载体编码GDNF(HSVgdnf)是否能保护神经元免受缺血性损伤。在原代神经元培养中，与对照载体HSVlac相比，HSVgdnf可减轻氧化剂诱导的损伤。在单侧大脑中动脉闭塞(MCA)60min之前或之后，向大脑皮层注射HSVgdnf或HSVlac，以测试体内的保护作用。对照组中风动物出现运动迟缓和运动不对称；HSVgdnf预处理显著减少了这种运动缺陷。在缺血损伤后接受HSVlac或HSVgdnf的动物没有表现出任何行为改善。卒中后一个月进行的组织学分析显示，接受HSVgdnf治疗的大鼠缺血组织丢失减少。同样，这些动物对胶质纤维酸性蛋白(GFAP)和caspase-3的免疫染色较少。综上所述，我们的数据表明HSVgdnf预处理对脑缺血具有保护作用，并支持利用HSV扩增子向中枢神经系统治疗输送营养因子。