GENE DELIVERY FOR SPIRAL CORD INJURY REPAIR

用于修复螺旋索损伤的基因传递

• 批准号: 6540174
• 负责人: Harold David Shine
• 金额: $ 16.23万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-15 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6540174
• 关键词: Adenoviridae; Animalia; gene therapy; genetic transduction; method development; nervous system regeneration; neuromuscular system; neuronal guidance; neurotrophic factors; spinal cord injury; transfection /expression vector

DESCRIPTION: (adapted from applicant's abstract) Treatment of spinal cord injury (SCI) encompasses the rescue of injured neurons, the promotion of axonal regeneration and the formation of functional synapses. This project focuses on one aspect of an envisioned multi-modai therapy - the promotion of axonal regeneration. The proposed experiments are designed to test the hypothesis that local release of induced neurotrophic factors (NFs) in the spinal cord will enhance axonal regeneration after trauma. When adenoviral vectors carrying NF genes (Adv-nf) are injected into peripheral muscles they are transported to motoneurons where the genes are expressed and NFs are released. The NFs protect spinal cord motoneurons from trauma-induced death without evoking an inflammatory response. To test the hypothesis that local expression of NFs will support axonal regeneration adenoviral vectors will be delivered to the spinal cord in which the corticospinal tract (CST) has been unilaterally lesioned at the level of the pyramids. In this lesion model there is no trauma in the spinal cord so that tissue destruction, ischemia, and paralysis are not factors in determining the outcome and may allow better understanding of the effects of NFs on sprouting. The specific aims of this proposal are to: Test whether expression of NFs by genetically modified cells in the spinal cord will induce and sustain axonal sprouting after injury (Specific aim 1). Genes for neurotrophin 3 (NT-3), nerve growth factor (NGF), ciliary neurotrophic factor (CNTF), brain-derived neurotrophic factor (BDNF) and glial cell line-derived growth factor (GDNF) will be delivered singly and in combination to the spinal cord by: (1) direct injection of Adv-nfs into spinal cord tissue, (2) transduction of ependymal cells so that they release NFs into the cerebrospinal fluid, and (3) retrograde delivery of Adv-nfs to dorsal root ganglion and motoneurons. A panel of adenoviral vectors will be generated that contain a FLAG sequence to aid in immunological detection and the Elongation Factor (EF) promoter that is more efficient in mammalian cells. Specific aim 2 is designed to characterize the extent of expression and the pattern of sprouting. Specific aim 3 is designed to characterize the effects of Adv-nf-induced sprouting on motor behavior. If the investigators demonstrate that this strategy is effective in inducing axonal growth it would be a basis of future studies leading to a treatment of SCI.

描述：（改编自申请人的摘要）脊髓的处理 受伤（SCI）涵盖了受伤的神经元的营救，促进轴突 再生和功能突触的形成。这个项目重点 设想的多摩托疗法的一个方面 - 促进轴突 再生。提出的实验旨在检验以下假设。 脊髓中局部释放诱导的神经营养因子（NFS）将 增强创伤后轴突再生。当载有NF的腺病毒载体 将基因（ADV-NF）注入到外围肌肉中 表达基因并释放NFS的运动神经元。 NFS保护 创伤引起的死亡中的脊髓运动神经元，而无需唤起 炎症反应。为了测试NF的局部表达的假设 支撑轴突再生腺病毒载体将被输送到脊柱 皮质脊髓道（CST）已单侧病变的绳索 金字塔的水平。在此病变模型中，没有创伤 脊髓，使组织破坏，缺血和瘫痪不是因素 在确定结果并可以更好地理解 NFS发芽。该提案的具体目的是：测试是否是否 脊髓中基因修饰细胞的NF表达将诱导 并在受伤后维持轴突发芽（特定目标1）。基因 神经营养蛋白3（NT-3），神经生长因子（NGF），睫状神经营养因子 （CNTF），脑衍生的神经营养因子（BDNF）和神经胶质细胞系衍生 生长因子（GDNF）将单独交付并组合到脊柱 电线：（1）直接注入ADV-NFS脊髓组织，（2） 室室室细胞的转导，使其释放NFS到脑脊液中 流体，（3）ADV-NF逆行递送到背根神经节和 运动神经元。将生成一组腺病毒载体，其中包含一个 标志序列有助于免疫检测和伸长因子（EF） 在哺乳动物细胞中更有效的启动子。特定目标2是设计的 表征表达程度和发芽模式。具体的 AIM 3旨在表征ADV-NF诱导的发芽的影响 运动行为。如果调查人员证明此策略是 有效诱导轴突生长，这将是未来研究的基础 导致SCI治疗。