LATENT HERPES VECTORS FOR NEURODEGENERATIVE DISORDERS

神经退行性疾病的潜伏疱疹载体

• 批准号: 3417311
• 负责人: JACK G STEVENS
• 金额: $ 34.97万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-30 至 1995-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3417311
• 关键词: Alzheimer's disease; Herpesviridae; Parkinson's disease; corpus striatum; experimental brain lesion; fibroblast growth factor; gene expression; growth factor receptors; hippocampus; immunocytochemistry; in situ hybridization; laboratory mouse; laboratory rat; latent virus infection; microdialysis; nervous system regeneration; neural degeneration; neurotrophic factors; recombinant DNA; retina degeneration; substantia nigra; transfection

Neurodegenerative diseases constitute a major human health problem. In three of these conditions, i.e. Alzheimer's disease and age-related retinal degeneration, specific neuronal populations die, and they have been shown to respond to neurotrophic factors other gene products. Major difficulties concerned with the use of these molecules are the methods of delivery. To promote e restoration of normal function within neural circuits, the methods should supply the products in the physiologic context (i.e., by the appropriate cells) and 2) provide the products for extended periods with minimal pathologic consequences. The technologies employed to date do not meet these standards. To deliver relevant genes, we have developed latent, nonpathogenic Herpes simplex virus vectors. These vectors are designed to express foreign proteins for extended periods, and they neither replicate nor reactivate from the latent state. With a prototype agent, we have demonstrated stable, long-term expression of a foreign gene m neurons of intact animal. We now propose to express those genes whose products have either been shown or strongly considered to alter progression of the diseases just mentioned. In the case of Alzheimer's disease, we will use vectors expressing NGF, BDNF and NT-3 and assess role of these factors in the regulation of central cholinergic circuits, their potential role in the pathogenesis of AD and their possible therapeutic role in two models of cholinergic deficiency, the fimbria-fornix lesion and the senile age rat. Virus will be inoculated into the hippocampus and basal forebrain, expression of the growth factors verified by in situ hybridization and, where possible, by immuno- cytochemistry. Then, the cholinergic response will be determined by immunostaining for NGF receptor, choline acetyltranferase and, in the case of the senile rat model, by behavioral studies. For Parkinson's disease, viruses encoding similar trophic factors and tyrosine hydroxylase (TH) will be introduced into the substantia nigra and corpus striatum of intact and chemically lesioned animals. The effect of the factors be assayed by staining for dopamine and TH, by microdialysis for extracellular dopamine, and by quantitation of apomorphine-induced rotation. For degeneration, vectors expressing basic fibroblast growth factor and the "retinal degeneration slow gene" will be injected Into retinae of animals with inherited and light-induced retinal degeneration, and the response will be monitored by direct observation of the retinal nuclear layer. These experiments will result in the development of powerful new tools for study of the nervous system and characterize the potential therapeutic role of growth factors and related molecules in neurodegenerative disorders.

神经退行性疾病是人类健康的一个主要问题。 在 其中三种病症，即阿尔茨海默氏病和与年​​龄相关的视网膜 退化，特定的神经元群体死亡，并且它们已被证明 对神经营养因子其他基因产物做出反应。 主要困难 与这些分子的使用相关的是递送方法。到 促进神经回路内正常功能的恢复， 方法应在生理背景下提供产品（即通过 适当的电池）和2）提供产品延长期限 最小的病理后果。 迄今为止所采用的技术不 满足这些标准。为了传递相关基因，我们开发了潜在的、 非致病性单纯疱疹病毒载体。 这些载体旨在 长时间表达外源蛋白，并且它们不复制 也不会从潜伏状态重新激活。 有了原型代理，我们就有了 证明外源基因在神经元中稳定、长期表达 完整的动物。 我们现在建议表达那些其产物具有 已被证明或强烈认为可以改变疾病的进展 刚才提到的疾病。 就阿尔茨海默病而言，我们将使用表达 NGF 的载体， BDNF 和 NT-3，并评估这些因素在中枢调节中的作用 胆碱能回路及其在 AD 发病机制中的潜在作用 它们在两种胆碱能缺乏模型中可能的治疗作用， 穹窿伞损伤和老年大鼠。 病毒将被接种 进入海马体和基底前脑，表达生长因子 通过原位杂交验证，并在可能的情况下通过免疫验证 细胞化学。 然后，胆碱能反应将由下式确定： NGF 受体、胆碱乙酰转移酶的免疫染色，在这种情况下 通过行为研究建立老年大鼠模型。对于帕金森病， 编码相似营养因子和酪氨酸羟化酶（TH）的病毒将 被引入完整的黑质和纹状体 受到化学损伤的动物。 各因素的影响可通过以下方式进行分析 通过微透析检测细胞外多巴胺，对多巴胺和 TH 进行染色， 以及通过定量阿朴吗啡引起的旋转。 对于退化， 表达碱性成纤维细胞生长因子和“视网膜 退化慢基因”将被注射到动物视网膜中 遗传性和光诱导性视网膜变性，反应将是 通过直接观察视网膜核层进行监测。这些 实验将导致强大的新研究工具的开发 神经系统并表征潜在的治疗作用 神经退行性疾病中的生长因子和相关分子。