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：该转化研究项目致力于解决与神经性疼痛相关的病理生理过程，以及慢性疼痛、自身免疫、传染病及其在人类患者中的交叉作用的潜在汇合。慢性神经性疼痛可以影响身体的任何部位，并且可能由于各种损伤、感染、自身免疫或代谢紊乱（例如糖尿病周围神经病变）而发生。我们正在测试这样的假设：在一些患者中，慢性疼痛是由与针对周围神经或雪旺细胞或可能的中枢神经系统成分中的蛋白质产生的自身抗体相关的免疫病理学过程引发和/或维持的。已知自身抗体是某些大纤维副肿瘤性周围神经病的罪魁祸首，大纤维也可能导致神经性感觉异常。如果疼痛是其中的一个组成部分，我们假设存在针对小直径疼痛感知（伤害性）C-或A-δ神经纤维的神经末梢中发现的蛋白质的自身抗体。为了检验疼痛性神经病具有自身免疫成分的假设，我们建立了一种灵敏的定量液相发光测定法，该测定法使用重组蛋白抗原-荧光素酶融合物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）人类慢性疼痛疾病的潜在分子和细胞生物学机制，以及（b）利用这些知识来设计针对疼痛和其他可适应的疾病的新治疗和诊断方法。 我们建立的方法，即荧光素酶免疫沉淀系统 (LIPS) 测定，可以稳健、灵敏地检测血清、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病症，包括有或没有神经系统受累的病毒和细菌感染，以及在部分患者中具有神经系统症状的自身免疫性疾病。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 等人，2016）。我们的目的是确定 LIPS 检测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（带状疱疹）和带状疱疹患者的研究，在这些患者中，带状疱疹演变成一种称为带状疱疹后神经痛（PHN）的痛苦神经系统疾病。我们在带状疱疹患者亚群中检测到一些中和性抗细胞因子自身抗体，有趣的是，所有具有抗体的患者都患有 PHN。这很重要，因为它表明一些 PHN 患者可能需要额外的干预来控制疾病，而不仅仅是镇痛药物。在最近的一篇出版物中，我们分别在死后人类三叉神经节和全血中证明了 HSV1 转录本和 HSV1 抗体。滴度与转录本的对应率为100%。事实上，我们使用 RNA-seq 使我们能够将 HSV1 读数与 HSV1 基因组对齐，并将所有转录本与 HSV1 潜伏相关转录本 (LAT) 对齐，这表明没有受试者在死亡。我们可以使用这种方法作为更大规模的头痛和偏头痛研究的一部分。 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渐进分层和演变。随着我们增加测试抗原的数量以及跨条件和疾病的检测，我们对免疫和自身免疫反应进行了综合评估。这是通过确定（a）对直系同源蛋白质和蛋白质片段的免疫反应的程度和特异性，（b）指示共同点或一般机制的抗原谱的重叠，以及（c）参与的整个信号传导途径内的抗原性来实现的。细胞间或细胞内通讯。最终，可以实施完整的多抗原分析，以更深入地了解许多复杂的人类疾病状态的免疫成分。我们最近的综述总结了我们目前对致病性自身抗体的看法。几乎所有自身抗体具有致病作用的自身免疫性疾病都表明抗体靶向分泌肽或蛋白质或完整质膜蛋白的胞外结构域。后者通常是跨膜信号蛋白。现在，我们可以在寻找抗原靶标时重点关注跨膜离子通道、受体和酶的细胞外结构域。这一概念极大地简化了对可能参与产生或维持慢性神经性疼痛病症的抗原的搜索。