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; 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.

项目摘要： 这项研究的目的是验证脊髓损伤后神经性疼痛的新治疗方法（SCI-NP） 使用新型且高度相关的猪模型。这项研究具有创新性，因为我们将验证令人兴奋的 啮齿动物研究的新发现，在临床上发现了科学NP的高效治疗方法 相关的猪模型。具体而言，新疗法涉及提供编码6份的基因构建体 NMDA拮抗剂丝氨酸 - 甲状腺素和1份阿片类激动剂内啡肽1（6shg/em1）通过 jergova等人，2017年所详细介绍的载体内递送。的确在研究中被验证，大鼠与 在研究期间（12周），6SHG/EM1基因疗法没有表现出异常性疾病的回归。数据也是如此 表明6SHG/EM1的作用通过鞘内注射抗SHG和纳洛酮瞬时阻断 表明影响是由于“靶向”机制所致。转基因在脊柱中的表达 在注射位置附近的背喇叭附近及其周围的神经元细胞中证实了绳索 （Jergova等，2017）。为了加速这种新疗法的翻译，我们有反翻译的定量 人类医学中使用的感觉测试（QST）方法用于评估用于猪使用的神经性疼痛。在 与QST测试的结合，我们还开发了猪疼痛量表，包括反身和 脊柱上的反应，对于疼痛研究中的翻译至关重要。我们将采用合作 测试总体假设的方法，即Sci后的6SHG/EM1在 一个非常相关的猪模型，密切模仿人类科学。我们的目标是优化和扩展此事 通过使用临床上的猪模型来定义所需的关键变量，因此高效的科幻治疗 成功翻译。我们将测试我们的总体假设，并通过 完成三个具体目标。在AIM 1中，我们将优化成功交付AAV的方法 猪模型中SCI后对脊髓的基因治疗。在AIM 2中，我们将检验以下假设： 在SCI之后的亚急性期间，在猪模型中施用6SHG/EM1。科幻 模型将是猪的胸腔挫伤/压缩​​损伤，我们的研究已经确定 实验室以及其他人使用的。在多个皮肤上的基线定量感觉测试（QST） 将进行。 QST包括评估热（冷热），触觉，压力和动态刺激。 疼痛反应将在我们开发的猪诱发疼痛量表上进行评分，其中包括 脊柱（情感）和脊柱（反射）成分。 SCI后第6周（神经性疼痛的时间点） 开发），猪将接受腺相关病毒（AAV）中6shg/em1构建体的单次注射。 电机和QST响应将在随后的12周内评估。在AIM 2中，我们将检验假设 SCI后的亚急性期间，SCI后的6SHG/EM1给药会引起SHG的表达和 EM1转基因和肽中的肽。 SCI后12周，来自AIM 2的猪将是人道的 提取的脊髓组织和脊髓组织，背根神经节和脑脊液。我们将评估 使用荧光 - 连接的免疫吸附测定（FLISA）和免疫组织化学评估。表达方式将是 与疼痛结果数据相关。这项研究的结果是翻译这一新的关键下一步 对临床试验的治疗，最终为患有SCI后神经性疼痛的退伍军人提供缓解。