Optogenetics to improve hand function after spinal cord injury.

光遗传学改善脊髓损伤后的手部功能。

• 批准号: 10252778
• 负责人: Polina O Anikeeva
• 金额: $ 64.48万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10252778
• 关键词: Acute; Address; Affect; Anatomy; Animal Model; Animals; Axon; Blood; Blood flow; Brain; Bypass; Cervical; Cervical spinal cord injury; Cervical spinal cord structure; Chronic; Clinical; Contusions; Data; Electric Stimulation; Electrodes; Electrophysiology (science); Exposure to; FOS gene; Fiber; Forelimb; Functional disorder; Genes; Goals; Hand; Histology; Immunohistochemistry; Implant; Implantation procedure; In Situ Hybridization; Injury; Ion Channel; Ischemia; Label; Light; Lighting; Mediating; Methods; Modeling; Movement; Nervous System Trauma; Neuroanatomy; Neurons; Neurostimulation procedures of spinal cord tissue; Opsin; Optics; Paralysed; Paresis; Polymers; Proteins; Randomized; Rattus; Recovery; Recovery of Function; Research; Respiratory physiology; Rodent; Site; Spinal; Spinal Cord; Spinal Cord Contusions; Spinal cord injury; Stimulus; Surface; Synapses; Technology; Testing; Training; Up-Regulation; Upper Extremity; Viral; Work; arm; arm function; axon growth; base; biomaterial compatibility; cell type; epidural space; experimental study; flexibility; hand rehabilitation; implantation; improved; improved functioning; injury recovery; innovation; mechanical properties; neural circuit; novel; optogenetics; recruit; relating to nervous system; response; treatment group

Project Summary/Abstract Restoration of hand and arm function is the highest treatment priority for people with cervical spinal cord injury. The goal of this research is to develop and test a novel method to improve recovery of hand and arm function after spinal cord injury. We propose to use optogenetic stimulation of the cervical spinal cord to both improve function and to uncover the mechanisms by which spinal cord stimulation leads to recovery. Our preliminary data demonstrate both a rapid and near complete recovery of forelimb function when animals receive optogenetic spinal cord stimulation following a clinically-realistic cervical spinal cord contusion injury. Optogenetic light stimulation may provide benefits by both directly activating neural circuits and also by increase blood flow to the injured spinal cord. Here we propose to compare the functional recovery resulting from optogenetic and electrical spinal cord stimulation, as well as the combination of electrical and light stimulation delivered to naïve animals that do not express optogenetic proteins. Our experiments are enabled by a novel multifunctional electrode that permits both optical and electrical stimulation to be delivered to the surface of the spinal cord in rodents. These flexible polymer electrodes will be refined in Aim 1 to deliver chronic optogenetic and epidural electrical stimulation to the rat cervical spinal cord. Thus all animals will be implanted with identical hardware prior to being randomized into treatment groups to permit a direct comparison between optogenetic and electrical stimulation in Aim 2. We will use our established rat model of spinal contusion injury where animals are trained to perform precision forelimb reaching to accurately quantify recovery of function after injury. Our collaborative team has developed a reliable method of viral transduction of optogenetic proteins such that light-sensitive ion channels are expressed in neurons of the non-transgenic rat cervical spinal cord. Following 6-weeks of treatment with optogenetic and epidural electrical stimulation, we will explore the mechanisms by which each treatment leads to prolonged recovery of forelimb function in Aim 3. We will perform terminal electrophysiology and record the responses evoked by both optical and electrical stimulation in the same animals. Our preliminary data demonstrate an upregulation of axon growth following optogenetic stimulation. We will use retrograde trans-synaptic tracing and histology to quantify new circuit formation bypassing the injury. Labelled neurons will be co-localized with the neurons activated by optogenetic vs. epidural stimulation using combined in-situ hybridization and immunohistochemistry to illuminate mechanisms of recovery. In summary, we propose to uncover the mechanisms by which optogenetic spinal cord stimulation leads to nearly complete recovery of forelimb function following spinal cord injury. Once understood, we expect these mechanism to directly inspire treatments for a range of neurological traumas to the brain and spinal cord.

项目摘要/摘要 恢复手和手臂功能是脊髓脊髓患者的最高治疗方法 受伤。这项研究的目的是开发和测试一种新的方法，以改善手臂的恢复 脊髓损伤后的功能。我们建议使用颈脊髓的光遗传学刺激 提高功能并发现脊髓刺激导致恢复的机制。 我们的初步数据表明，当 临床现实的脊髓挫伤后，动物接受光遗传学的脊髓刺激 受伤。光遗传刺激可以通过直接激活神经回路和通过 增加流向受伤的脊髓的血液。在这里，我们建议比较结果的功能恢复 来自光遗传学和电脊髓刺激，以及电气和光的组合 刺激传递给不表达光遗传蛋白的幼稚动物。 我们的实验是通过允许光学和电气的新型多功能电极来启用的 刺激要输送到啮齿动物中的脊髓表面。这些柔性聚合物电极将是 在AIM 1中进行了精制，以向大鼠颈脊髓传递慢性光学遗传学和硬膜外电刺激。 将所有动物都植入相同的硬件之前，然后将其随机分为治疗组 允许在AIM 2中的光遗传学和电刺激之间进行直接比较。 我们将使用我们已建立的脊柱挫伤大鼠模型，其中训练动物进行训练 精度前肢能够准确地量化受伤后功能的恢复。我们的协作团队有 开发了一种可靠的光学遗传蛋白转导病毒转导方法，使光敏离子通道 在非转基因大鼠宫颈脊髓的神经元中表达。遵循6周的治疗 光遗传学和硬膜外电刺激，我们将探索每种处理导致的机制 为了长期恢复AIM 3中的前肢功能。我们将执行末端电生理学并记录 同一动物的光学刺激和电刺激引起的反应。 我们的初步数据表明，光遗传刺激后轴突生长的上调。我们将 使用逆行反射突触跟踪和组织学来量化绕过损伤的新电路形成。 标记的神经元将与使用光遗传学与硬膜外刺激激活的神经元共定位 合并的原位杂交和免疫组织化学阐明了恢复机制。 总而言之，我们建议发现光遗传学脊髓刺激导致的机制 脊髓损伤后几乎完全恢复前肢功能。一旦理解，我们期望这些 直接激发针对大脑和脊髓一系列神经创伤的治疗的机制。