AAV MEDIATED NEUROTROPHIC FACTOR GENE DELIVERY TO BRAIN

AAV 介导的神经营养因子基因递送至大脑

• 批准号: 6187144
• 负责人: EDWIN M MEYER
• 金额: $ 20.33万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6187144
• 关键词: adeno associated virus group; animal old age; behavior test; brain septal area; cell type; cholinergic agents; denervation; gamma aminobutyrate; gene dosage; gene expression; gene therapy; genetic transduction; green fluorescent proteins; hippocampus; laboratory rat; memory; neurotrophic factors; phosphopyruvate hydratase; transfection; transfection /expression vector

The goal of this revised grant continues to be testing several hypotheses about neurotropic factor somatic gene transfer into the septohippocampal system. These studies use adeno-associated virus (AAV) recombinant vectors containing the neuron specific enolase (NSE) promoter to drive expression of nerve growth factor (NGF) or brain derived neurotrophic factor (BDNF). Initial studies demonstrating robust expression of the marker gene green fluorescent protein (GFP) in septum and hippocampus for extended intervals after transduction have been extended to observations about trophic factor expression, levels of cholinergic markers and trk-receptor density in this pathway. Expression is neuro-specific in each region studied. Some degree of tropism for subpopulations o neurons is also apparent in hippocampus, though not so in septum. Our grant proposes to test: 1) whether there are dose- and time-dependent expressions of NSE-driven GFP expression in different hippocampal and septal neurons: 2) if ectopic BDNF and NGF expression in intact septum and hippocampus elevates cholinergic and trk markers without altering GABAergic activity; 3) if BDNF and NGF gene transfer into septum preferentially protects axotomized cholinergic neurons in a manner not seen with either similar gene transfer in hippocampus or with GABAergic neurons: 4) whether both hippocampal and septal trophic factor gene transfer can improve memory-related behaviors in animals receiving partial fornix lesions; 5) whether age-related deficits in memory-related behaviors and acetylcholine release can be overcome with septal NGF or BDNF gene transfer; and 6) if aging interferes with the expression of transgenic NGF more than that of BDNF, as predicted from studies of endogenous trophic factor regulation during senescence. These studies may lead eventually to gene therapy trials for conditions associated with septohippocampal cholinergic deficits such as Alzheimer's disease.

这项修订后的拨款的目标是继续测试有关神经营养因子体细胞基因转移到隔海马系统的几个假设。 这些研究使用含有神经元特异性烯醇化酶 (NSE) 启动子的腺相关病毒 (AAV) 重组载体来驱动神经生长因子 (NGF) 或脑源性神经营养因子 (BDNF) 的表达。 初步研究表明，标记基因绿色荧光蛋白 (GFP) 在转导后在隔膜和海马中长时间稳定表达，现已扩展到对该途径中的营养因子表达、胆碱能标记水平和 trk 受体密度的观察。 每个研究区域的表达都是神经特异性的。 海马体中神经元亚群的某种程度的趋向性也很明显，但在隔膜中则不然。 我们的资助计划测试：1）不同海马和隔膜神经元中 NSE 驱动的 GFP 表达是否存在剂量和时间依赖性表达：2）完整隔膜和海马中的异位 BDNF 和 NGF 表达是否会升高胆碱能和 trk 标记而不改变 GABA 活性； 3）如果BDNF和NGF基因转移到隔膜中优先保护轴突性胆碱能神经元，其方式在海马或GABA能神经元中没有类似的基因转移：4）海马和隔膜营养因子基因转移是否可以改善接受部分穹窿损伤的动物的记忆相关行为； 5)是否可以通过间隔NGF或BDNF基因转移来克服与年龄相关的记忆相关行为和乙酰胆碱释放缺陷； 6) 衰老对转基因 NGF 表达的干扰是否大于对 BDNF 的干扰，正如衰老过程中内源营养因子调节研究所预测的那样。 这些研究可能最终导致针对与隔海马胆碱能缺陷相关的疾病（例如阿尔茨海默病）的基因治疗试验。