Enhancing Plasticity in Damaged Spinal Cord to Repair Transmission and Function

增强受损脊髓的可塑性以修复传输和功能

• 批准号: 8543006
• 负责人: Victor L Arvanian
• 金额: --
• 依托单位: NORTHPORT VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8543006
• 关键词: Accounting; Action Potentials; Acute; Address; Adult; Afghanistan; Anatomy; Animal Model; Animals; Award; Axon; Behavior; Biological Preservation; Cells; Chest; Chondroitin ABC Lyase; Chondroitin Sulfate Proteoglycan; Chronic; Cicatrix; Clinical; Clinical Trials; Combined Modality Therapy; Conflict (Psychology); Confocal Microscopy; Contusions; Demyelinations; Digestion; Electron Microscopy; Electrophysiology (science); Engineering; Enzymes; Exhibits; Experimental Models; Exposure to; FDA approved; Fiber; Fibroblasts; Freedom; Funding; Gene Delivery; Genes; Goals; Growth; Herpesvirus 1; Hindlimb; Home environment; Immunochemistry; Individual; Injury; Interneurons; Intramuscular; Intrathecal Injections; Ion Channel; Iraq; Lateral; Left; Lesion; Locomotor Recovery; Magnetism; Mediating; Mental disorders; Methods; Modeling; Molecular; Motor; Motor Neurons; Muscle; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; NR2D NMDA receptor; NTF3 gene; Natural regeneration; Neurons; Pathway interactions; Physiological; Physiology; Procedures; Property; Ranvier' s Nodes; Rattus; Recombinants; Recovery; Recovery of Function; Research; Scientist; Sensory; Side; Site; Soldier; Spinal; Spinal Cord; Spinal Cord Lesions; Spinal Injections; Spinal Injuries; Spinal cord damage; Spinal cord injury; Synapses; Synaptic plasticity; Therapeutic; Thoracic spinal cord structure; Time; Tissues; Transgenes; Translating; Treatment Efficacy; United States; Viral Vector; War; Work; adeno-associated viral vector; axon growth; base; behavior test; coping; disability; dorsal column; effective therapy; gene therapy; improved; in vivo; inhibitor/antagonist; motor function improvement; motor function recovery; multidisciplinary; nervous system disorder; neurotrophic factor; news; novel; operation; public health relevance; receptor function; remyelination; repaired; research study; response; spinal cord and brain injury; spinal tract; spine bone structure; therapy design; transmission process; white matter; Accounting; Action Potentials; Acute; Address; Adult; Afghanistan; Anatomy; Animal Model; Animals; Award; Axon; Behavior; Biological Preservation; Cells; Chest; Chondroitin ABC Lyase; Chondroitin Sulfate Proteoglycan; Chronic; Cicatrix; Clinical; Clinical Trials; Combined Modality Therapy; Conflict (Psychology); Confocal Microscopy; Contusions; Demyelinations; Digestion; Electron Microscopy; Electrophysiology (science); Engineering; Enzymes; Exhibits; Experimental Models; Exposure to; FDA approved; Fiber; Fibroblasts; Freedom; Funding; Gene Delivery; Genes; Goals; Growth; Herpesvirus 1; Hindlimb; Home environment; Immunochemistry; Individual; Injury; Interneurons; Intramuscular; Intrathecal Injections; Ion Channel; Iraq; Lateral; Left; Lesion; Locomotor Recovery; Magnetism; Mediating; Mental disorders; Methods; Modeling; Molecular; Motor; Motor Neurons; Muscle; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; NR2D NMDA receptor; NTF3 gene; Natural regeneration; Neurons; Pathway interactions; Physiological; Physiology; Procedures; Property; Ranvier' s Nodes; Rattus; Recombinants; Recovery; Recovery of Function; Research; Scientist; Sensory; Side; Site; Soldier; Spinal; Spinal Cord; Spinal Cord Lesions; Spinal Injections; Spinal Injuries; Spinal cord damage; Spinal cord injury; Synapses; Synaptic plasticity; Therapeutic; Thoracic spinal cord structure; Time; Tissues; Transgenes; Translating; Treatment Efficacy; United States; Viral Vector; War; Work; adeno-associated viral vector; axon growth; base; behavior test; coping; disability; dorsal column; effective therapy; gene therapy; improved; in vivo; inhibitor/antagonist; motor function improvement; motor function recovery; multidisciplinary; nervous system disorder; neurotrophic factor; news; novel; operation; public health relevance; receptor function; remyelination; repaired; research study; response; spinal cord and brain injury; spinal tract; spine bone structure; therapy design; transmission process; white matter

DESCRIPTION (provided by applicant): Many soldiers return home with contusive spinal cord injuries (SCI), mostly due to the powerful explosives used in the improvised bombs that rattle U.S. troops inside heavily armored vehicles. In fact, "One in Six Wounded in Afghanistan War Have Spinal Cord Injuries" (The Spinal Post, Nov.11, 2009"). Currently, there is no cure and consequently studies using animal models of SCI are imperative. Results of recent studies, including our own, revealed that there are at least three major factors known to limit functional recovery from incomplete SCI in adults: (1) the presence of axonal growth/regeneration inhibitors associated with the glial scar, (2) the lack of neurotrophin support, and (3) the decreased excitability and plasticity in pre- existing pathways to neurons in damaged spinal cord. In attempts to improve synaptic connectivity in the damaged spinal cord and examine if this will facilitate recovery of motor function after SCI, we have recently utilized a model of unilateral hemisection (HX) of the spinal cord in adult rats. This is convenient experimental model because one side of the cord is lesioned and other remains intact. Using this HX model we have recently developed (through studies funded by our initial Merit Award; 2008-2012), a novel additive treatment designed to neutralize axonal growth/ regeneration inhibitors, deliver neurotrophins and enhance plasticity in damaged spinal cord. We found that this combination treatment (in contrast to treatment with any single component) can re-establish novel synaptic connections around a lesion in chronically hemisected spinal cord. These physiological findings were supported by the observation of increased regeneration and branching of axons near the injury and better functional recovery. Components of this combination treatment are: (i) degradation of scar-associated inhibitory molecules (CSPGs) with the enzyme chondroitinase-ABC (ChABC), (ii) delivery of neurotrophin NT-3 (administered via engineered fibroblasts) and increased function of NMDA receptors in interneurons and motoneurons (using HSV1 viral vector-mediated expression of NMDA receptor NR2D regulatory subunits). The goal of the current proposal is to translate these principles into a clinically amenable application. The proposed research will be a continuation of our previous work, extended to a more realistic contusive SCI model, with refinement of methods for NT-3 delivery and enhanced spinal cord excitability that are FDA- approved or under clinical trials. This novel triple combination approach will combine Ch-ABC treatment with (i) AAV-mediated transgene delivery of NT-3 (AAV-NT3; intraspinal and intrathecal injections; AAV-mediated gene delivery has been found to be safe in multiple clinical trials) and (ii) repetitive electro-magnetic stimulation applied over intact spinal vertebrae (FDA-approved and widely used for treatment of neurological and psychiatric disorders). Due to increasing numbers of individuals with chronic SCI, we will utilize both acute and chronic mid-thoracic contusion SCI models in adult rats. To evaluate the efficacy of these treatments, we will collaborate with leading scientists in the field and conduct a multidisciplinary examination, including in vivo physiology, anatomy, immunochemistry, electron microscopy and behavior. Using these methods we will study the additive or synergistic effects of the triple therapeutic treatment on (1) strengthening synaptic connections through the injury epicenter to lumbar motoneurons, and then to hindlimb muscles (electrophysiology); (2) anatomical plasticity of fibers accounting for the persistence of the synaptic response after exposure to this novel treatment (anatomical tracing and confocal microscopy); (3) cellular and molecular mechanisms underlying the beneficial effects of the combination treatment (electron microscopy and immunochemisctry to study axon remyelination and distribution of ion channels); and (4) recovery of locomotor function in multiple behavioral tests. Preliminary results of on-going experiments show significant improvements of motor function in rats that have received the novel triple treatment compared to controls after thoracic contusive SCI.

描述（由申请人提供）： 许多士兵因脊髓损伤（SCI）返回家园，这主要是由于即兴炸弹中使用的强大炸药，这些炸弹在重装甲车辆中摇晃着美洲部队。实际上，“阿富汗战争中六分之一的受伤者脊柱受伤”（脊柱哨所，2009年11月11日”）。目前，尚无治愈方法，因此使用SCI动物模型进行了研究。最近的研究结果。最近的研究的结果，包括我们自己的研究，至少有三个主要因素限制了与不满的axte级相关的SCI（1）（1）（1）（1）（1）。 （2）缺乏神经营养蛋白的支撑，（3）在脊髓受损的神经元中的兴奋性和可塑性下降，以改善受损的脊髓中的突触连通性，并检查SCI后的运动是否促进了一项单一的模型，因为这是否促进了运动型模型。绳索的侧面是病变的，而其他您最近开发的HX模型（通过我们最初的优点奖励的研究； 2008- 2012年），一种新型的添加剂处理，旨在中和轴突生长/再生抑制剂，提供神经营养蛋白并增强受损脊髓的可塑性。我们发现，这种组合处理（与任何单个成分的治疗相反）可以重新建立围绕慢性半脊髓病变的新型突触连接。这些生理发现得到了观察到损伤附近轴突的再生和分支的增加，并获得了更好的功能恢复。这种组合处理的组成部分是：（i）与软骨软骨蛋白酶-ABC（CHABC）降解相关的疤痕相关抑制性分子（CSPG），（ii）神经营养蛋白NT-3的递送（通过工程成纤维细胞和使用NMDA的功能增加）（通过增加的NMDA受体函数）（用于施用） NMDA受体NR2D调节亚基的载体介导的表达）。 当前建议的目的是将这些原则转化为临床上可正常的应用。拟议的研究将是我们以前的工作的延续，扩展到了更现实的Contively Sci模型，并改进了NT-3递送的方法，并增强了FDA-批准或在临床试验下的脊髓兴奋性。这种新型的三重组合方法将将CH-ABC处理与（i）AAV介导的NT-3（AAV-NT3; AAV-NT3；次脑内和鞘内注射； AAV介导的基因输送）的转基因递送相结合，在多个临床试验中是安全的，用于多次临床试验和（ii）重复的电磁刺激（ii）跨性别的脊柱疗法（II），并促进了脊柱效应（FD AAP）（FD AAPARE）（FDACARETARITY and f. fd aa apaper）（FD AACINALITY）。精神病）。由于慢性SCI的个体数量的增加，我们将在成年大鼠中同时利用急性和慢性中胸腔挫伤SCI模型。为了评估这些治疗的功效，我们将与该领域的主要科学家合作 并进行多学科检查，包括体内生理学，解剖学，免疫化学，电子显微镜和行为。使用这些方法，我们将研究三重治疗治疗对（1）通过损伤震中加强突触连接的累加或协同作用，然后再对后肢肌肉（电生理学）来加强突触连接； （2）纤维的解剖学可塑性，该纤维在暴露于这种新型治疗后的突触反应持续性（解剖学跟踪和共聚焦显微镜）； （3）组合处理的有益作用（电子显微镜和免疫化学研究以研究离子通道的轴突再生和分布）的有益作用的基础的细胞和分子机制； （4）在多个行为测试中恢复运动功能。与胸腔cococi后的对照相比，正在进行的实验的初步结果显示，与对照组相比，接受了新型三重治疗的大鼠运动功能的显着改善。