CATIONIC VESICLES FOR BRAIN NGF GENE DELIVERY

用于脑 NGF 基因传递的阳离子囊泡

• 批准号: 6201000
• 负责人: J HUGHES
• 金额: $ 15.3万
• 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: 1999-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6201000
• 关键词: Alzheimer's disease; acidity /alkalinity; amyloid proteins; astrocytes; biotransformation; brain cell; cations; choline acetyltransferase; drug screening /evaluation; enzyme activity; gene expression; gene therapy; genetically modified animals; hippocampus; laboratory mouse; laboratory rat; liposomes; microinjections; neurotrophic factors; northern blottings; pharmacokinetics; plasmids; surfactant; tissue /cell culture; transfection /expression vector

The overall aim of this research is to use non-viral liposomal delivery systems as direct gene transfer vectors to study the efficacy of nerve growth factor in the therapy of Alzheimer's disease (AD) pathogenesis. Neurotrophic factors (viz., nerve growth factor [NGF]) exogenous delivery to the aged brain may prevent neuronal degeneration. NGF's large molecular weight preludes easy permeation across the blood brain barrier. Consequently, in order to obtain therapeutic effects, the peptide must be directly administered to the brain by using non-viral gene delivery methods for expressing NGF, brain tissue should secrete NGF in a biological active form permitting chronic, well tolerated and regionally specific NGF production. In AD, this approach could be utilized to retard cholinergic neuronal degeneration. Specifically, non-viral gene delivery vectors will be developed to optimize gene expression in brain tissue after direct injection. Three variables will be investigated in the optimization of this delivery system. Initially, the addition of an endosomal membrane rupturing agent which will increase the amount of nucleic acid reaching the cell's cytoplasm will be tested. The second objective will focus on the inclusion of nuclear peptide localization sequences to facilitate the transfer of vector to the nucleus. A third component involves the synthesis of cationic lipids which will form smaller, enzymatic-resistive DNA-complexes leading to enhanced distribution and cellular uptake within the brain. After optimization of the liposomal delivery vehicle in vitro, it will be used to test several hypotheses on the influence of NGF-gene transfer into the brain. The influence of gene transfer on the cholinergic system, NGF- receptor up regulation, and memory-related behaviors will be explored using two model systems: axotomy of septal cholinergic neurons and the aging rat. These studies are expected to demonstrate that NGF-gene delivery can protect cholinergic neurons and improve behavior in hypofunctional animals, and that the newly developed gene-transfer vehicles are superior to present techniques based on reconstituted virosomes.

这项研究的总体目的是使用非病毒脂质体递送 系统作为直接基因传递向量研究神经的功效 阿尔茨海默氏病（AD）发病机理治疗的生长因子。 神经营养因子（即神经生长因子[NGF]）外源递送 对老年大脑可能会预防神经元变性。 NGF的大分子 体重在整个血脑屏障上易于渗透。 因此，为了获得治疗作用，肽必须为 通过使用非病毒基因递送方法直接给大脑施用 为了表达NGF，脑组织应分泌NGF 允许慢性，耐受性良好和特定区域的NGF的形式 生产。 在AD中，可以利用这种方法来阻碍胆碱能 神经元变性。 具体而言，将开发非病毒基因递送向量以优化 直接注射后脑组织中的基因表达。 三个变量 将在此输送系统的优化中进行研究。 最初，添加内体膜破裂剂将 增加到达细胞细胞质的核酸的量将是 测试。 第二个目标将重点放在核的包含上 肽定位序列，以促进向量转移到 核。 第三个成分涉及阳离子脂质的合成 将形成较小的酶促抗DNA复合物，从而增强 大脑内的分布和细胞摄取。 在体外优化脂质体递送车后，它将是 用于检验关于NGF基因转移到中的影响的几个假设 大脑。 基因转移对胆碱能系统NGF-的影响 受体UP调节和与内存有关的行为将使用 两个模型系统：间隔胆碱能神经元和老化大鼠的轴突切开术。 预计这些研究将证明NGF-Gene的交付可以 保护胆碱能神经元并改善动物功能性动物的行为， 并且新开发的基因转移车辆优越 基于重构病毒体的技术。