Evaluation of 6SHG/EM1 as a treatment for spinal cord injury-induced neuropathic pain in a pig model

6SHG/EM1 治疗猪模型脊髓损伤引起的神经性疼痛的评价

• 批准号: 10935563
• 负责人: CANDACE L. FLOYD
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10935563
• 关键词: Acceleration; Affective; Anatomy; Animal Model; Animals; Back; Biological Assay; Blood; Caring; Cells; Cerebrospinal Fluid; Characteristics; Chest; Clinical; Clinical Trials; Collection; Contusions; Data; Dependovirus; Development; Disease; Documentation; Dose; Euthanasia; Evaluation; Family suidae; Gene Delivery; Genes; Goals; Human; Hypersensitivity; Immune response; Immunosorbents; Impairment; Injections; Injury; Intrathecal Injections; Investigational New Drug Application; Laboratory Research; Life; Link; Location; Medical; Medicine; Methodology; Methods; Modeling; Motor; N-Methylaspartate; Naloxone; Neurons; Opioid agonist; Pain; Pain Research; Pathology; Patients; Pattern; Peptides; Persons; Physiology; Positioning Attribute; Prevalence; Rattus; Reporting; Research; Rodent; Sensory; Serine; Seroprevalences; Spinal; Spinal Cord; Spinal Cord Lesions; Spinal Ganglia; Spinal Injections; Spinal cord injury; Stimulus; Tactile; Testing; Therapeutic; Time; Tissues; Transfection; Transgenes; Translating; Translations; United States Food and Drug Administration; Validation; Vertebral column; Veterans; Veterans Health Administration; adeno-associated viral vector; allodynia; antagonist; chronic neuropathic pain; chronic pain; clinically relevant; collaborative approach; delivery vehicle; dermatome; dorsal horn; efficacious treatment; endomorphin 1; experience; experimental study; first-in-human; gene therapy; good laboratory practice; innovation; negative affect; neutralizing antibody; novel; novel therapeutics; pain outcome; pain scale; painful neuropathy; porcine model; pressure; response; scale up; transgene expression; translational goal; vector

Project Summary: The goal of this research is to validate a novel treatment for neuropathic pain after spinal cord injury (SCI-NP) using a novel and highly clinically-relevant pig model. This research is innovative as we will validate exciting new findings from research in rodents which discovered a highly efficacious treatment for SCI-NP in a clinically- relevant pig model. Specifically, the new therapy involves delivering a gene construct encoding 6 copies of the NMDA antagonist serine-histrogranin and 1 copy of the opioid agonist endomorphin 1 (6SHG/EM1) via intraspinal delivery as detailed in Jergova et al., 2017. Indeed in the study to be validated, rats with the 6SHG/EM1 gene therapy did not exhibit a return of allodynia for the duration of the study (12 weeks). Data also showed that the effects of 6SHG/EM1 were transiently blocked by intrathecal injection of anti-SHG and naloxone, suggesting that the effects were due to "on-target" mechanisms. The expression of the transgenes in the spinal cord was confirmed in neuronal cells near and around the dorsal horn in the vicinity of the injection location (Jergova et al., 2017). To accelerate translation of this new therapy, we have back-translated quantitative sensory testing (QST) methods used in human medicine to evaluate neuropathic pain for use in pigs. In conjunction with QST testing, we have also developed a pig pain scale that includes both reflexive and supraspinal responses, critically important for translation in pain research. We will employ a collaborative approach to test the overarching hypothesis that administration of 6SHG/EM1 after SCI ameliorates SCI-NP in a highly clinically-relevant pig model which closely mimics human SCI. Our goal is to optimize and scale-up this highly efficacious SCI-NP treatment by using a clinically-relevant pig model to define key variables needed for successful translation. We will test our overarching hypothesis and achieve these translational goals by accomplishing three specific aims. In aim 1 we will optimize the methodology for successful delivery of AAV gene therapy to the spinal cord after SCI in a pig model. In aim 2 we will test the hypothesis that post-SCI administration of 6SHG/EM1 in the sub-acute period after SCI will ameliorate SCI-NP in a pig model. The SCI model will be a midthoracic contusion/compression injury in pigs that is well established in our research laboratory and as well as used by others. Baseline quantitative sensory testing (QST) over multiple dermatomes will be conducted. QST includes assessments of thermal (hot and cold), tactile, pressure, and dynamic stimuli. Pain responses will be scored on the porcine evoked pain scale that we developed which includes both supra- spinal (affective) and spinal (reflexive) components. At weeks 6 after SCI (a time point when neuropathic pain is developed), pigs will receive intraspinal injection of 6SHG/EM1 construct in adeno-associated virus (AAV). Motor and QST responses will be evaluated for a subsequent 12 weeks. In aim 2, we will test the hypothesis that post-SCI administration of 6SHG/EM1 in the sub-acute period after SCI induces expression of SHG and EM1 transgenes and peptides in the spinal cord. At 12 weeks after SCI, pigs from aim 2 will be humanely euthanized and spinal cord tissue, dorsal root ganglia, and cerebrospinal fluid extracted. We will assess the presence and expression pattern of SHG and EM1 transgenes and peptides in these tissues using fluorescent- linked immunosorbent assays (FLISA) and immunohistochemical evaluations. Expression patterns will be correlated with pain outcome data. The results of this study are the critical next step in translation of this new therapy to clinical trials, and ultimately to provide relief to Veterans who suffer from neuropathic pain after SCI.

项目概要： 这项研究的目的是验证一种治疗脊髓损伤后神经性疼痛 (SCI-NP) 的新方法 使用一种新颖且高度临床相关的猪模型。这项研究具有创新性，因为我们将验证令人兴奋的 啮齿类动物研究的新发现，在临床上发现了 SCI-NP 的高效治疗方法 相关猪模型。具体来说，新疗法涉及传递编码 6 个拷贝的基因构建体。 NMDA 拮抗剂丝氨酸组织粒蛋白和 1 份阿片类激动剂内吗啡肽 1 (6SHG/EM1)，来自 Jergova 等人，2017 年详细介绍了椎管内递送。事实上，在待验证的研究中，大鼠 6SHG/EM1 基因治疗在研究期间（12 周）没有表现出异常性疼痛的复发。数据还 结果表明，鞘内注射抗 SHG 和纳洛酮可暂时阻断 6SHG/EM1 的作用， 表明这种影响是由于“目标”机制造成的。转基因在脊髓中的表达 在注射位置附近的背角附近和周围的神经元细胞中证实了绳索 （杰戈瓦等人，2017）。为了加速这种新疗法的转化，我们反向翻译了定量 人类医学中使用的感觉测试（QST）方法，用于评估猪的神经性疼痛。在 结合 QST 测试，我们还开发了猪疼痛量表，其中包括反射性和 脊髓上反应对于疼痛研究的转化至关重要。我们将聘请一个协作 检验总体假设的方法，即 SCI 后给予 6SHG/EM1 可改善 SCI-NP 一种与临床高度相关的猪模型，与人类 SCI 非常相似。我们的目标是优化和扩大规模 通过使用临床相关的猪模型来定义治疗所需的关键变量，从而实现高效的 SCI-NP 治疗 翻译成功。我们将测试我们的总体假设并通过以下方式实现这些转化目标： 实现三个具体目标。在目标 1 中，我们将优化成功交付 AAV 的方法 猪模型脊髓损伤后的脊髓基因治疗。在目标 2 中，我们将检验以下假设：后 SCI 在 SCI 后的亚急性期给予 6SHG/EM1 将改善猪模型中的 SCI-NP。 SCI 模型将是我们研究中已确立的猪胸中部挫伤/压缩​​损伤 实验室以及其他人使用。对多个皮区进行基线定量感官测试 (QST) 将进行。 QST 包括热（热和冷）、触觉、压力和动态刺激的评估。 疼痛反应将根据我们开发的猪诱发疼痛量表进行评分，其中包括超 脊髓（情感）和脊髓（反射）成分。 SCI 后第 6 周（神经性疼痛发生的时间点） 已开发），猪将接受腺相关病毒（AAV）中的 6SHG/EM1 构建体的脊髓内注射。 将在接下来的 12 周内评估运动和 QST 反应。在目标 2 中，我们将检验假设 SCI 后亚急性期给予 6SHG/EM1 诱导 SHG 表达 脊髓中的 EM1 转基因和肽。 SCI 后 12 周，目标 2 中的猪将受到人道对待 安乐死并提取脊髓组织、背根神经节和脑脊液。我们将评估 使用荧光-在这些组织中 SHG 和 EM1 转基因和肽的存在和表达模式 连锁免疫吸附测定（FLISA）和免疫组织化学评估。表达模式将是 与疼痛结果数据相关。这项研究的结果是这一新成果转化的关键下一步 治疗到临床试验，并最终为 SCI 后遭受神经性疼痛的退伍军人提供缓解。