Gene Replacement to Activate Spinal Axon Regeneration

基因替换激活脊髓轴突再生

• 批准号: 6435031
• 负责人: J H PATE SKENE
• 金额: $ 34.65万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-12-15 至 2004-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6435031
• 关键词: Adenoviridae; actin binding protein; axon; embryo /fetus tissue /cell culture; gene therapy; genetically modified animals; laboratory mouse; laboratory rat; nervous system regeneration; neural growth associated protein; nonhuman therapy evaluation; recombinant virus; sciatic nerve; spinal cord injury; transfection /expression vector

DESCRIPTION (provided by applicant): Damage to the spinal cord results in permanent loss of sensation and motor control below the level of injury, because the long axons that connect the spinal cord to the brain fail to regenerate. This contrasts starkly with the vigorous regrowth of axons in injured peripheral nerves, where successful regeneration is closely correlated with the re-induction of many genes ordinarily expressed during development. Spinal cord lesions often fail to activate those genes, and the injured cells are unable to regenerate their axons. Using transgenic mice, we have found that expression of just two of those genes - coding for the growth cone proteins growth associated protein (GAP)-43 and cytoskeleton associated protein (CAP)-23 -- triggers a 60-fold increase in the ability of adult neurons to support regeneration of spinal cord axons in vivo. The small number of proteins required to activate regeneration raises the possibility that viral-mediated expression or other forms of direct gene replacement in injured neurons could become one key component of future strategies to stimulate spinal axon repair. The proposed studies constitute a test in principle of this strategy. We first will determine which combination(s) of GAP-43, CAP-23, and the related protein myristoylated alanine-rich C kinase substrate (MARCKS) are most effective in triggering the intrinsic ability of neurons to support axon regeneration, and the conditions under which viral-mediated expression of these genes can activate this growth competence after spinal cord or dorsal root injury in normal (non-transgenic) adult animals. The second aim is to identify specific contexts in which expression of GAP-43, CAP-23 and/or MARCKS may overcome the effects of growth-inhibiting molecules in the spinal cord environment, and conversely to identify those inhibitory elements that continue to impair axon extension by neurons expressing GAP-43, CAP-23 and MARCKS. Together, the proposed studies outline the essential features of a new strategy for overcoming one key hurdle to central nervous system (CNS) axon repair, and provide a guide to additional treatments that would need to be combined with this gene therapy approach in order to obtain effective regeneration of spinal axons in vivo.

描述（由申请人提供）：脊髓的损害导致 在伤害水平以下的永久感觉和电动机控制的永久丧失， 因为将脊髓连接到大脑的长轴突无法 再生。这与轴突的剧烈再生长形成鲜明对比 受伤的周围神经，成功再生密切相关 随着在发育过程中通常表达的许多基因的重新诱导。 脊髓病变通常无法激活这些基因，而受伤的细胞 无法再生其轴突。使用转基因小鼠，我们发现 仅表达了其中两个基因 - 编码生长锥蛋白 生长相关蛋白（GAP）-43和细胞骨架相关蛋白（CAP）-23 - 触发成年神经元支持的能力增加了60倍 体内脊髓轴突的再生。少数蛋白质 激活再生所需的可能性增加了病毒介导的可能性 受伤神经元中的表达或其他形式的直接基因替代可能 成为刺激脊柱修复的未来策略的关键组成部分。 拟议的研究原则上构成了该策略的测试。我们首先 将确定GAP-43，CAP-23和相关蛋白的组合组合 肉豆蔻酰化富含丙氨酸的C激酶底物（MARCKS）最有效 触发神经元支持轴突再生的内在能力，并 病毒介导的这些基因表达的条件可以 激活脊髓或背根损伤后这种生长能力 正常（非转基因）成年动物。第二个目的是确定具体 GAP-43，CAP-23和/或MARCK的表达的上下文可能会克服 脊髓环境中抑制生长分子的影响， 相反，可以识别那些继续损害轴突的抑制元素 表达GAP-43，CAP-23和MARCKS的神经元扩展。在一起， 拟议的研究概述了新策略的基本特征 克服中枢神经系统（CNS）轴突修复的一个关键障碍，并 为需要与之结合的其他治疗提供指南 这种基因治疗方法是为了获得有效的脊柱再生 轴突在体内。