Ischemic Spasticity: Modulation by GAD65 Gene Delivery

缺血性痉挛：GAD65 基因传递的调节

• 批准号: 7426389
• 负责人: MARTIN MARSALA
• 金额: $ 33.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7426389
• 关键词: Acute; Address; Adverse effects; Agonist; Animals; Ankle; Aortic Aneurysm; Area; Attention; Baclofen; Brain; Cannulations; Cells; Chronic; Clinical; Clinical Management; Confocal Microscopy; Dependovirus; Development; Dyskinetic syndrome; Electromyography; Engineering; Fiber; Fluorescence; GABA-B Receptor; Gene Delivery; Gene Expression; Gene Transfer; Genes; Goals; H-Reflex; Immunofluorescence Immunologic; Immunohistochemistry; Injection of therapeutic agent; Interneurons; Intervention; Intrathecal Injections; Ischemia; Lead; Measures; Mediating; Microdialysis; Modality; Modeling; Motor; Motor Neurons; Muscle; Muscle Rigidity; Neurons; Numbers; Parkinsonian Disorders; Patients; Perfusion; Peripheral; Pharmacological Treatment; Population; Proteins; Rattus; Research Personnel; Resistance; Rotation; Spinal; Spinal Cord; Spinal Ganglia; Spinal Injuries; Spinal cord injury; Staging; Subfamily lentivirinae; Symptoms; Therapeutic; Time; Treatment Efficacy; Up-Regulation; Viral Vector; extracellular; fusion gene; gamma-Aminobutyric Acid; indexing; inhibitory neuron; novel therapeutics; programs; receptor; receptor expression; relating to nervous system; repaired; research study; size; therapeutic gene; transgene expression

DESCRIPTION (provided by applicant): Using a rat spinal ischemia model, we have shown that specific periods of transient spinal ischemia lead to a selective degeneration of small and medium-sized inhibitory neurons in lumbosacral segments and the development of prominent and permanent spasticity and rigidity. Similar development of spasticity and rigidity in patients undergoing thoracoabdominal aortic aneurysm repair or after traumatic spinal injury had been described. While the clinical signs of spasticity and rigidity can be effectively controlled by systemic or spinal treatment with several pharmacological agents including baclofen (GABA B receptor agonist) or tizadinine (alpha2 agonist), significant side effects and complications associated with chronic intrathecal cannulations calls for the development of new and more effective therapies. In recent years, significant attention has been focused on a potential use of adeno-or lenti-viral vectors with the goal to upregulate therapeutic genes in specific areas of the brain or spinal cord. These initial studies clearly show that intrastriatal injections of adeno-associated virus encoding GAD65 gene is effective in suppressing parkinsonian dyskinesias and is associated with an increase in local GABA release. In the present studies, using an established rat model of ischemic spasticity and rigidity, we will examine a possible therapeutic potential of spinal lentivirus-mediated GAD65 gene delivery in order to produce local upregulation of GABA synthesis and ameliorate spasticity and rigidity. The effect of local GAD65 gene upregulation in animals with developed spasticity and rigidity will be accomplished and assessed by i) direct spinal parenchymal lentivirus delivery or spinal grafting of rat neuronal precursors genetically modified to produce GABA, ii) electrophysiological/functional indices of spasticity and rigidity, iii) corresponding spinal parenchyma! GABA release, and, iv) histological analysis of transgene expression. From a practical standpoint, these studies will systematically address issues which we believe have both basic and clinical importance. Thus the ability to selectively increase GABA synthesis in previously ischemic, GABA neuron-depleted segments, may prove to be of particular significance in developing novel therapeutic modalities for managing chronic spasticity and rigidity in patients after ischemic or traumatic spinal injury.

描述（由申请人提供）：使用大鼠脊髓缺血模型，我们已经表明，特定时期的短暂脊髓缺血导致腰骶段中小型和中型抑制性神经元的选择性变性，以及显著和永久性痉挛和僵硬的发展。在接受胸腹主动脉瘤修复术或创伤性脊柱损伤后的患者中，痉挛和僵硬的发展也有类似的描述。虽然痉挛和僵硬的临床体征可以通过用包括巴氯芬（GABA B受体激动剂）或替扎地宁（α 2激动剂）的几种药理学药剂进行全身或脊柱治疗来有效地控制，但是与慢性鞘内插管相关的显著副作用和并发症需要开发新的和更有效的疗法。近年来，腺病毒或慢病毒载体的潜在用途受到了极大的关注，其目的是上调脑或脊髓特定区域中的治疗基因。这些初步研究清楚地表明，纹状体内注射编码GAD 65基因的腺相关病毒可有效抑制帕金森病运动障碍，并与局部GABA释放增加有关。在本研究中，使用一个建立的大鼠模型缺血性痉挛和僵硬，我们将研究脊髓慢病毒介导的GAD 65基因传递，以产生局部上调GABA的合成和改善痉挛和僵硬的可能的治疗潜力。将通过以下方式实现和评估具有发展的痉挛和僵硬的动物中局部GAD 65基因上调的作用：i）直接脊髓实质慢病毒递送或经遗传修饰以产生GABA的大鼠神经元前体的脊髓移植，ii）痉挛和僵硬的电生理/功能指数，iii）相应的脊髓实质慢病毒递送或脊髓移植。GABA释放，和，iv）转基因表达的组织学分析。从实践的角度来看，这些研究将系统地解决我们认为具有基础和临床重要性的问题。因此，选择性增加GABA合成的能力，在以前缺血，GABA神经元耗尽的部分，可能被证明是特别重要的开发新的治疗模式，用于管理慢性痉挛和僵硬的患者缺血性或创伤性脊髓损伤后。