GENE THERAPY OF BASAL FOREBRAIN CHOLINERGIC LESIONS

基底前脑胆碱能病变的基因治疗

• 批准号: 6372429
• 负责人: LEON J THAL
• 金额: $ 27.51万
• 依托单位: VETERANS MEDICAL RESEARCH FDN/SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6372429
• 关键词: Alzheimer's disease; acetylcholine; choline; choline acetyltransferase; disease /disorder model; experimental brain lesion; gene therapy; laboratory rat; neurochemistry; nonhuman therapy evaluation

DESCRIPTION: (adapted from applicant's abstract) Lesions of the cholinergic basal forebrain (CBF) system in animals using an immunotoxin produce robust behavioral deficits and mimic some aspects of Alzheimer's disease (AD). This model is useful for testing both the role of acetylcholine in cognitive processes and new therapeutic strategies such as gene therapy. The ability to control gene expression is an important requirement of this technology for clinical application. The vector systems being explored in this proposal may be exogenously regulated. This proposal will investigate mechanisms of restoring neurotransmitter function in animals with CBF immunotoxin lesions by 1) enhancing ACh release postgrafting via choline supplementation, and 2) transplanting cells capable of repressing or inducing the transgene for choline acetyltransferase (ChAT). The repressible system (tetracycline), and the inducible system (eccdysteroid), modulates gene expression of ChAT in a rapid, reversible and highly specific fashion. In the first series of experiments, the investigators will determine whether different doses of exogenously administered choline can augment the production and release of acetylcholine from genetically engineered fibroblasts grafted to the cortex and hippocampus of rats following immunotoxic lesions of the CBF. Once an increase in release has been determined, behavioral effects of this augmented release will be determined. For the second and third series of experiments, they will use the regulatable cell lines that either induce or repress ChAT expression after administration of doxycycline or Muristerone A, respectively. In their immunotoxin lesion they will demonstrate that acetylcholine released from these cells after grafting to the neocortex and hippocampus is both necessary and sufficient for behavioral recovery. The effects of these regulatable genes will be tested in a spatial and non-spatial task after determining the dose and duration of the exogenous compound needed to induce or repress gene expression. If successful, the experiments will demonstrate that release of ACh can be exogenously controlled in animals and results in significant behavioral improvement. The control of transmitter release is necessary before grafting or direct gene insertion experiments can be entertained in humans in order to be able to safely terminate delivery of the gene product. These approaches will not only be applicable for AD, but other neurodegenerative disorders and for delivering other transgenes of interest.

描述：（改编自申请人的摘要）胆碱能损伤 动物的基底前脑（CBF）系统使用免疫毒素产生强大的 行为缺陷并模仿阿尔茨海默病（AD）的某些方面。这 该模型可用于测试乙酰胆碱在认知中的作用 过程和新的治疗策略，例如基因治疗。有能力 控制基因表达是该技术的重要要求 临床应用。本提案中正在探索的载体系统可能是 外源调节。该提案将研究恢复机制 CBF 免疫毒素损伤动物的神经递质功能 1) 通过补充胆碱增强移植后乙酰胆碱释放，2) 移植能够抑制或诱导胆碱转基因的细胞 乙酰转移酶（ChAT）。可抑制系统（四环素）和 诱导系统（蜕皮类固醇），以快速、 双面且高度特定的时尚。 在第一个系列的实验中，研究人员将确定是否 不同剂量的外源性给予胆碱可以增加产量 移植到的基因工程成纤维细胞释放乙酰胆碱 CBF 免疫毒性损伤后大鼠的皮质和海马。 一旦确定释放增加，这种行为的影响 将确定增加释放。 对于第二和第三系列的实验，他们将使用可调节的 给药后诱导或抑制 ChAT 表达的细胞系 分别为多西环素或Muristerone A。在他们的免疫毒素损伤中，他们 将证明移植后这些细胞释放乙酰胆碱 新皮质和海马对于行为来说既是必要的又是充分的 恢复。这些可调控基因的影响将在空间中进行测试 确定外源性药物的剂量和持续时间后的非空间任务 诱导或抑制基因表达所需的化合物。如果成功的话， 实验将证明乙酰胆碱的释放可以被外源控制 并导致动物行为显着改善。 嫁接或直接基因前必须控制递质释放 可以在人类身上进行插入实验，以便能够 安全地终止基因产物的递送。这些方法不仅将 适用于 AD，但也适用于其他神经退行性疾病和分娩 其他感兴趣的转基因。