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.

项目总结： 本研究的目的是验证一种治疗脊髓损伤后神经病理性疼痛(sci-np)的新方法。 使用一种新的、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 对啮齿动物的研究发现了一种治疗SCI-NP的高效方法，这种疗法在临床上是有效的- 相关的生猪模型。具体地说，新的治疗方法包括传递编码6个拷贝的 NMDA拮抗剂丝氨酸-组氨酸颗粒素和1拷贝阿片激动剂内吗啡1(6SHG/EM1)通过 椎管内分娩，详见Jergova等人，2017年。事实上，在有待验证的研究中，患有 6SHG/EM1基因治疗在研究期间(12周)没有表现出痛觉异常的复发。数据也 鞘内注射抗SHG抗体和纳洛酮可暂时阻断6SHG/EM1的作用。 这表明，这种影响是由于“对准目标”的机制造成的。转基因在脊髓中的表达 脊髓在注射部位附近的背角附近和周围的神经细胞中得到证实。 (Jergova等人，2017年)。为了加速这种新疗法的翻译，我们有回译的量化 人类医学中用于评估猪神经病理性疼痛的感觉测试(QST)方法。在……里面 结合QST测试，我们还开发了一种猪疼痛量表，包括反射性和 脊椎上的反应，对于疼痛研究中的翻译至关重要。我们将采用协作式 脊髓损伤后应用6SHG/EM1改善SCI-NP的总体假说的检验方法 一种高度临床相关的猪模型，非常接近于人类脊髓损伤。我们的目标是优化和扩展这一功能 使用与临床相关的猪模型定义所需的关键变量进行高效的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翻译目标 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 猪脊髓损伤后脊髓基因治疗的实验研究。在目标2中，我们将检验后SCI的假设 亚急性期应用6SHG/EM1可改善猪脊髓损伤后的脊髓损伤-NP。《SCI》 我们的研究已经建立了猪中胸挫伤/压迫伤的模型。 实验室，也可供他人使用。多个皮肤体的基线定量感觉测试(QST) 将会进行。QST包括对热(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，该量表包括以上两个方面- 脊椎(情感)和脊柱(反射)成分。在脊髓损伤后6周(神经病理性疼痛的时间点 )，猪将接受6SHG/EM1腺相关病毒(AAV)构建的脊髓内注射。 运动和QST反应将在随后的12周内进行评估。在目标2中，我们将检验假设 脊髓损伤后亚急性期应用6SHG/EM1诱导SHG和 脊髓中的EM1转基因和多肽。在脊髓损伤后12周，来自AIM 2的猪将是人道的 安乐死并提取脊髓组织、背根神经节和脑脊液。我们将评估 用荧光法检测SHG和EM1转基因及多肽在这些组织中的存在和表达模式 联合免疫吸附试验(FLISA)和免疫组织化学检测。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的翻译的关键下一步 治疗到临床试验，并最终为遭受脊髓损伤后神经病理性疼痛的退伍军人提供缓解。