GENE DELIVERY TO THE CNS TO MODULATE EPILEPSY

CNS疾病的常规药物治疗，包括手术和 药理学上，通常对大脑区域有广泛的不良影响， 参与治疗的疾病。针对这一点，我们 发展出我们称之为“分子神经外科”的技术。分子 神经外科是一种治疗神经系统疾病的新方法， 基因改造病毒来调节神经系统细胞。 立体定向局部递送允许调节地理上定义的 而病毒设计可以允许细胞特异性 和大分子的表达。 该项目的长期目标是提供一个工程化的 编码谷氨酸脱羧酶（GAD）的转录单位（转基因） 到培养物中的原代神经细胞和大脑中的离散位置， 通过缺陷型单纯疱疹病毒（HSV）载体。拟将这一 将增加γ-氨基丁酸（GABA）的生物合成和/或 增加谷氨酸降解。一组新的GAD转基因载体将 将允许检测和表征 体内转基因产物。体外实验可以让我们 表征GAD转基因表达的程度及其对 GABA和谷氨酸在包括神经胶质细胞在内的各种细胞类型中的代谢 以及多巴胺能和GABA能神经元。我们将测试我们的能力， 使用大鼠复杂部分脑缺血模型调节体内GABA合成 癫痫发作，“区域tempestas”。将GAD转基因体内递送至 在梨状前皮质深部（区域）中的一个关键的触发癫痫的位点 tempestas）或GABA介导的抗惊厥作用的关键部位， 中脑（黑质）将使我们能够评估功能 这种定点基因治疗的影响。 这些技术将 用于癫痫和GABA/谷氨酸的基础神经生物学研究 新陈代谢.最终，它们将形成一种新形式的 癫痫治疗和作为其他分子神经外科的模型 紊乱超过200万美国人患有癫痫，其中 大约25%的患者对药物治疗难治， 手术治疗 其他神经系统疾病也适用于 分子神经外科学包括神经变性疾病、遗传性疾病、神经变性疾病。 诸如亨廷顿舞蹈症、疼痛调节和生化- 基于精神病的疾病。该提案结合了以下技能： 神经外科用于高度局部化的应用，分子生物学用于 缺陷疱疹载体的创建，以及建模的药理学 癫痫发作和局部与全身调节相结合的研究。

Conventional medical therapies for CNS disorders, both surgical and pharmacological, often have widespread unwanted effects on brain areas not involved in the disorder being treated. In response to this we are developing what we have termed "molecular neurosurgery". Molecular neurosurgery is a novel approach to nervous system disorders that uses genetically modified viruses to modulate cells in the nervous system. Stereotactic local delivery permits modulation of a geographically defined cell population whereas the viral design can allow for cell specificity and expression of large molecules. The long term goal of this project is to deliver an engineered transcription unit (transgene) encoding glutamic acid decarboxylase (GAD) to primary neural cells in culture and to discrete locations in the brain, via defective herpes simplex virus (HSV) vectors. It is proposed that this will increase the biosynthesis of gamma-aminobutyric acid (GABA) and/or increase glutamate degradation. A novel set of GAD transgene vectors will be constructed that will permit detection and characterization of the transgene product in vivo. The in vitro experiments will allow us to characterize the extent of GAD transgene expression and its influence on the metabolism of GABA and glutamate in various cell-types including glia and glutaminergic and GABAergic neurons. We will test our ability to modulate GABA synthesis in vivo using a rat model of complex partial seizures, the "area tempestas". In vivo delivery of the GAD transgene into a crucial seizure-triggering locus in the deep prepiriform cortex (area tempestas) or into a key site of GABA-mediated anticonvulsant action in the midbrain (substantia nigra) will allow us to evaluate the functional impact of this site-directed gene therapy. These techniques will be useful for basic neurobiological studies of epilepsy and GABA/glutamate metabolism. Eventually, they will form the basis for a new form of epilepsy treatment and as a model for molecular neurosurgery of other disorders. Over 2 million Americans suffer from epilepsy of which approximately 25% are refractory to drug treatment and are candidates for surgical therapy. Other nervous system disorders also amenable to molecular neurosurgery include neurodegenerative disorders, genetic disorders such as Huntington's chorea, pain modulation and biochemically- based psychiatriC disorders. This proposal combines the skills of neurosurgery for highly localized application, molecular biology for the creation of defective herpes vectors, and pharmacology for the modeling of epileptic seizures and studies of combined local and systemic modulation.