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）胆碱能病变 使用免疫毒素的动物中的基础前脑（CBF）系统可产生强大的 行为缺陷和模仿阿尔茨海默氏病（AD）的某些方面。这 模型对于测试乙酰胆碱在认知中的作用很有用 过程和新的治疗策略，例如基因治疗。能力 控制基因表达是该技术的重要要求 临床应用。该提案中正在探索的向量系统可能是 外源调节。该建议将调查恢复的机制 CBF免疫毒素病变的动物中神经递质的功能1） 通过补充胆碱增强ACH释放后的释放，2） 移植细胞能够抑制或诱导胆碱的转基因 乙酰转移酶（CHAT）。可抑制系统（四环素）， 诱导系统（Eccdysteroid），调节聊天的基因表达 可逆且高度具体的时尚。 在第一个实验中，研究人员将确定是否 不同剂量的外源施用胆碱可以增加产量 并从嫁接的基因工程成纤维细胞中释放乙酰胆碱 CBF免疫毒性病变后大鼠的皮质和海马。 一旦确定了发布的增加，这一点的行为影响 将确定增强释放。 对于第二和第三系列实验，它们将使用可调节的 诱导或压抑聊天表达后的细胞系 分别是强力霉素或肉豆蔻酮A。在他们的免疫毒素病变中 将证明乙酰胆碱嫁接后从这些细胞释放 新皮层和海马既需要行为必要且足够 恢复。这些可调节基因的影响将在空间中进行测试 确定外源的剂量和持续时间后，非空间任务 诱导或抑制基因表达所需的化合物。如果成功， 实验将证明可以外源控制ACH的释放 在动物中，导致行为改善。 在移植或直接基因之前，必须控制发射机释放 可以在人类中进行插入实验，以便能够 安全终止基因产品的递送。这些方法不仅会 适用于AD，但其他神经退行性疾病并提供 其他感兴趣的转基因。