Mechanisms of Pain and Immune Processes

疼痛和免疫过程的机制

• 批准号: 10487162
• 负责人: Andrew Mannes
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10487162
• 关键词: Acute; Acute Pain; Adaptive Immune System; Address; Affect; Afferent Neurons; Analgesics; Animal Model; Animals; Antibodies; Antigens; Autoantibodies; Autoantigens; Autoimmune; Autoimmune Diseases; Autoimmune Responses; Autoimmunity; Autopsy; Bacterial Infections; Biological; Biological Assay; Biological Factors; Biological Markers; Body part; C Fiber; Caliber; Cell Surface Proteins; Cell membrane; Cells; Cerebrospinal Fluid; Cessation of life; Chemotherapy-induced peripheral neuropathy; Chickenpox; Chronic; Clinical Research; Communicable Diseases; Communication; Complex; Data; Data Set; Diabetes Mellitus; Diagnostic; Diagnostic tests; Disease; Disease susceptibility; Dysesthesias; Enzymes; Evaluation; Evolution; Extracellular Domain; Fiber; Ganglia; Generations; Genome; Goals; Headache; Herpes zoster disease; Herpesviridae; Herpesvirus 1; Herpesvirus Type 3; Human; Immune; Immune response; Immunology; Immunoprecipitation; Infection; Inflammation; Injury; Intervention; Investigation; Ion Channel; Knowledge; Life; Liquid substance; Luciferases; Maintenance; Mammalian Cell; Measures; Membrane Proteins; Metabolic Diseases; Methodology; Migraine; Modeling; Molecular; Nature; Nerve; Nerve Endings; Nerve Fibers; Nervous system structure; Neuraxis; Neurobiology; Neurons; Neuropathy; Nociception; Operative Surgical Procedures; Pain; Pain Disorder; Paraneoplastic Polyneuropathy; Pathogenicity; Pathologic; Patients; Peptides; Peripheral Nerves; Peripheral Nervous System Diseases; Pharmaceutical Preparations; Phase; Plasma; Play; Postherpetic neuralgia; Predictive Value; Process; Production; Protein Fragment; Proteins; Publications; Recombinant Proteins; Recording of previous events; Research; Role; Saliva; Sampling; Schwann Cells; Serum; Signal Pathway; Signaling Protein; Specificity; Spinal Cord; Spinal cord damage; Structure of trigeminal ganglion; System; Technology; Testing; Thinking; Time; Tracer; Transcript; Transmembrane Domain; Traumatic CNS injury; Traumatic injury; Viral; Virus Diseases; Whole Blood; antibody detection; antigen test; base; chemotherapeutic agent; chronic neuropathic pain; chronic pain; chronic pain patient; chronic painful condition; cytokine; diabetic; diagnostic biomarker; disorder control; drug development; extracellular; human disease; insight; latency associated transcript; luminescence; nerve injury; nervous system disorder; painful neuropathy; pathogenic autoantibodies; patient subsets; receptor; relating to nervous system; tissue injury; tool; transcriptome; transcriptome sequencing; translational research program; virology

Overview: This translational research program addresses pathophysiological processes related to neuropathic pain and the potential confluence of chronic pain, autoimmunity, infectious diseases, and their intersection in human patients. Chronic neuropathic pain can affect any part of the body and can occur due to a variety of insults, infections, autoimmune or metabolic disorders (e.g., diabetic peripheral neuropathy). We are testing the hypothesis that, in some patients, chronic pain is initiated and/or maintained by immunopathological processes related to autoantibodies generated against proteins in peripheral nerve or Schwann cells or possibly components of the central nervous system. Autoantibodies are known culprits in certain large fiber paraneoplastic peripheral neuropathies and large fibers can also contribute to neuropathic dysesthesias. Where pain is a component, we hypothesize the presence of autoantibodies to proteins found in nerve endings arising from small diameter pain-sensing (nociceptive) C- or A-delta nerve fibers. To test the hypothesis that painful neuropathic conditions have an autoimmune component, we established a sensitive, quantitative, liquid phase luminescence assay that uses recombinant protein antigen-luciferase fusions as tracers obtained after expression in mammalian cells. The goals of this research are to understand (a) potential molecular and cell biological mechanisms underlying human chronic pain disorders, and (b) to use this knowledge to devise new treatments and diagnostics for pain and other disorders to which it can be adapted. The methodology we established, the luciferase immunoprecipitation systems (LIPS) assay, robustly and sensitively detects antibodies in serum, plasma, cerebrospinal fluid, or saliva. Our early investigations evaluated a range of diseases and disorders that antibodies play a role in, including viral and bacterial infections with and without nervous system involvement and autoimmune disorders that have nervous system symptomology in subsets of patients. In many neural autoimmune disorders, the major autoantigens are frequently plasma membrane receptors or ion channels (Burbelo et al., 2016). Our aim is to determine if the LIPS assay can detect auto-antibodies to these proteins in appropriate chronic pain patients. For example, we completed a study on shingles (herpes zoster) and patients in which shingles evolved into a painful neurological disorder called post-herpetic neuralgia (PHN). We detected some neutralizing anti-cytokine autoantibodies in a subpopulation of Zoster patients, and, interestingly, all of the patients with antibodies had PHN. This is important because it suggests that some patients with PHN may require additional intervention to control the disorder other than just analgesic medications. In a recent publication we demonstrated both HSV1 transcripts and HSV1 antibodies in post-mortem human trigeminal ganglion and whole blood, respectively. The correspondence between titer and transcript was 100%. The fact that we used RNA-seq allowed us to align the HSV1 reads to the HSV1 genome and all of the transcripts aligned with the HSV1 latency associated transcript (LAT), indicating that none of the subjects had viral re-activation at the time of death. We can use this approach as a component in a larger study of headache and migraine. One of the most compelling aspects of this project is its broad applicability and the progressive layering and evolution of the datasets. As we increase the number of test antigens, and assay across conditions and diseases, we assemble comprehensive evaluations of immune and autoimmune responses. This is accomplished by determination of (a) the extent and specificity of immune response to orthologous proteins and protein fragments, (b) overlap in antigen profiles indicative of common denominators or general mechanisms, and (c) antigenicity within an entire signaling pathway involved in inter- or intracellular communication. Eventually, full multiple antigen profiling can be implemented to obtain a deeper level of understanding of the immune component of many complex human disease states. Our recent review summarizes our current thinking on pathogenic autoantibodies. Nearly all autoimmune disorders for which the autoantibody has a pathogenic action show that the antibodies target either a secreted peptide or protein or the extracellular domain of an integral plasma membrane protein. The latter are usually transmembrane signaling proteins. We can now focus on the extracellular domains of transmembrane ion channels and receptors and enzymes in the search for antigenic targets. This concept greatly simplifies the search for antigens that may participate in generating or sustaining a chronic neuropathic pain condition.

概述：该转化研究计划介绍了与神经性疼痛以及慢性疼痛，自身免疫性，传染病，传染病及其在人类患者中的相交的潜在汇合有关的病理生理过程。慢性神经性疼痛会影响身体的任何部位，并且可能由于各种侮辱，感染，自身免疫性或代谢性疾病（例如糖尿病性周围神经病）而发生。我们正在检验以下假设：在某些患者中，慢性疼痛是通过与外周神经或schwann细胞中蛋白质或中枢神经系统中可能成分的蛋白质产生的自身抗体有关的免疫病理过程和/或维持的。在某些大型纤维副塑性外周神经病中，自身抗体是已知的罪魁祸首，并且大型纤维也可以有助于神经性发抖。在疼痛是成分的地方，我们假设存在自身抗体与由小直径疼痛感（伤害感受）C-或A-二尔特神经纤维引起的神经末端发现的蛋白质。为了测试疼痛神经性疾病具有自身免疫性成分的假设，我们建立了一种灵敏的，定量的，液相发光测定法，该测定使用重组蛋白抗原葡萄糖酶融合作为在哺乳动物细胞中表达后获得的示踪剂。这项研究的目标是了解（a）人类慢性疼痛障碍的潜在分子和细胞生物学机制，以及（b）使用这些知识来设计新的治疗方法和诊断疼痛和其他可以适应的疾病。 我们确定的方法，即荧光素酶免疫沉淀系统（LIPS）测定，稳健而敏感地检测血清，血浆，脑脊液或唾液中的抗体。我们的早期研究评估了抗体在其中发挥作用的一系列疾病和疾病，包括病毒和细菌感染，有或没有神经系统介入和自身免疫性疾病，这些疾病和自身免疫性疾病在患者亚群中具有神经系统症状。在许多神经自身免疫性疾病中，主要的自身抗原通常是质膜受体或离子通道（Burbelo等，2016）。我们的目的是确定嘴唇测定是否可以在适当的慢性疼痛患者中检测到这些蛋白质的自身抗体。例如，我们完成了一项关于带状疱疹（疱疹带状疱疹）的研究和木瓦演变成一种疼痛的神经系统疾病，称为疗程后神经痛（PHN）。我们检测到带状疱疹患者亚群中的一些中和抗周围动物自身抗体，有趣的是，所有抗体患者均均为PHN。这很重要，因为它表明某些PHN患者可能需要额外的干预措施来控制这种疾病，而不仅仅是镇痛药。在最近的出版物中，我们分别证明了验尸后人类三叉神经节和全血的HSV1转录本和HSV1抗体。滴度和转录本之间的对应关系为100％。我们使用RNA-seq的事实使我们能够对齐HSV1读取与HSV1基因组以及与HSV1潜伏期相关转录本（LAT）一致的所有转录本，这表明在死亡时，没有一个受试者在死亡时具有病毒重新激活。我们可以将这种方法用作更大的头痛和偏头痛研究中的组成部分。 该项目最引人注目的方面之一是其广泛的适用性以及数据集的逐步分层和演变。随着我们增加测试抗原的数量，以及跨疾病和疾病的测定，我们会汇总对免疫和自身免疫反应的全面评估。这是通过确定（a）对直系同源蛋白和蛋白质片段的免疫反应的程度和特异性来实现的，（b）在抗原特征中重叠，指示了共同的分母或一般机制，以及（c）涉及与细胞间或细胞内通信的整个信号通路内的抗原性。最终，可以实施完整的多种抗原分析，以更深入地了解许多复杂人类疾病状态的免疫成分。我们最近的评论总结了我们目前对病原自身抗体的思考。自身抗体具有致病作用的几乎所有自身免疫性疾病都表明，抗体靶向分泌的肽或蛋白质或整体质膜蛋白的细胞外域。后者通常是跨膜信号蛋白。现在，我们可以专注于跨膜离子通道的细胞外结构域，以及在寻找抗原靶标时的受体和酶。这个概念大大简化了可能参与产生或维持慢性神经性疼痛状况的抗原的搜索。