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: 10237552
• 负责人: CANDACE L. FLOYD
• 金额: --
• 依托单位: VA SALT LAKE CITY HEALTHCARE SYSTEM
• 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/10237552
• 关键词: Acute; Affective; Anatomy; Animal Model; Animals; Back; Biological Assay; Blood; Caring; Cells; Cerebrospinal Fluid; Characteristics; Clinical; Clinical Trials; Collection; Contusions; Data; Dependovirus; Development; Disease; Documentation; Dose; 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; Rat-1; Rattus; Reflex action; 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; 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; spinal reflex; 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.

项目摘要： 本研究的目的是验证一种新的治疗脊髓损伤后神经病理性疼痛（SCI-NP）的方法 使用一种新型的高度临床相关的猪模型。这项研究是创新的，因为我们将验证令人兴奋的 啮齿动物研究的新发现，在临床上发现了一种非常有效的SCI-NP治疗方法， 猪的相关模型具体地说，新疗法涉及递送编码6个拷贝的 NMDA拮抗剂丝氨酸-组构颗粒蛋白和1个拷贝的阿片激动剂内吗啡肽1（6SHG/EM 1）通过 脊柱内递送如Jergova等人所述，2017.事实上，在有待验证的研究中， 6SHG/EM 1基因治疗在研究期间（12周）没有表现出异常性疼痛的恢复。数据还 结果表明，鞘内注射抗SHG抗体和纳洛酮可短暂阻断6SHG/EM 1的作用， 这表明这些影响是由于“靶向”机制。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索 （Jergova等人，2017年）。为了加速这种新疗法的转化，我们已经将定量 感觉测试（QST）方法用于人类医学中以评估用于猪的神经性疼痛。在 结合QST测试，我们还开发了一个猪疼痛量表，包括反射和 脊髓上的反应，在疼痛研究中的翻译至关重要。我们将聘请一个合作 一种检验SCI后给予6SHG/EM 1改善SCI-NP的总体假设的方法， 一种高度临床相关的猪模型，其非常类似于人类SCI。我们的目标是优化和扩大这一点， 通过使用临床相关的猪模型来定义所需的关键变量， 成功的翻译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，我们将优化成功递送AAV的方法。 在猪模型中对SCI后的脊髓进行基因治疗。在目标2中，我们将测试SCI后 在SCI后的亚急性期施用6SHG/EM 1将改善猪模型中的SCI-NP。的SCI 模型将是在我们的研究中建立的猪中胸挫伤/压迫损伤 实验室和其他人使用。多个皮区的基线定量感觉测试（QST） 将进行。QST包括热（热和冷），触觉，压力和动态刺激的评估。 疼痛反应将在我们开发的猪诱发疼痛量表上进行评分，该量表包括超- 脊髓（情感）和脊髓（反射）成分。在SCI后6周（神经性疼痛发生的时间点）， 开发），猪将接受腺相关病毒（AAV）中的6SHG/EM 1构建体的脊柱内注射。 将在随后的12周内评价运动和QST反应。在目标2中，我们将检验假设 在SCI后的亚急性期给予6SHG/EM 1诱导SHG的表达， 脊髓中的EM 1转基因和肽。在SCI后12周，将对来自aim 2的猪进行人道处理。 安乐死并提取脊髓组织、背根神经节和脑脊液。我们将评估 使用荧光标记的SHG和EM 1转基因和肽在这些组织中的存在和表达模式。 联合免疫吸附试验（弗里萨）和免疫组织化学评价。表达模式将是 与疼痛结果数据相关。这项研究的结果是关键的下一步翻译这一新的 从治疗到临床试验，并最终为SCI后遭受神经性疼痛的退伍军人提供缓解。