GENE DELIVERY TO THE CNS TO MODULATE EPILEPSY

中枢神经系统疾病的传统药物治疗，包括外科和 药理作用，往往对大脑区域有广泛的有害影响 与正在接受治疗的精神障碍有关。作为对此的回应，我们正在 发展我们所称的“分子神经外科”。分子 神经外科是一种治疗神经系统疾病的新方法，它使用 用来调节神经系统细胞的转基因病毒。 立体定向本地递送允许调制地理上定义的 细胞群，而病毒设计可以允许细胞特异性 以及大分子的表达。 该项目的长期目标是提供一个经过设计的 编码谷氨酸脱羧酶的转录单位(转基因) 培养中的原代神经细胞和大脑中离散的位置， 通过有缺陷的单纯疱疹病毒(HSV)载体。据建议，这一点 将增加γ-氨基丁酸(GABA)和/或的生物合成 增加谷氨酸的降解。一组新的GAD转基因载体将 被构造为允许检测和表征 体内转基因产物。体外实验将使我们能够 表征GAD转基因表达的程度及其对 GABA和谷氨酸在包括胶质细胞在内的不同细胞类型中的代谢 以及谷氨酰胺能和GABA能神经元。我们将测试我们的能力 用大鼠复合部分性激素模型调节体内GABA的合成 癫痫发作，即“区域风暴”。GAD转基因基因在体内的传递 深部梨状皮质(区)中一个关键的癫痫发作触发部位 或进入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 这种定点基因疗法的影响。这些技术将是 适用于癫痫和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 癫痫的治疗和作为其他分子神经外科的典范 精神错乱。超过200万美国人患有癫痫，其中 大约25%的人对药物治疗无效，并适合 外科治疗。其他神经系统疾病也容易患上 分子神经外科包括神经退行性疾病，遗传性 亨廷顿舞蹈症、疼痛调制和生化-- 以精神障碍为基础。这项建议结合了 神经外科的高度局部化应用，分子生物学的应用 创建有缺陷的疱疹病毒载体，以及用于建模的药理学 癫痫发作和局部和全身联合调节的研究。

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.