Mechanisms of Pain and Immune Processes

疼痛和免疫过程的机制

• 批准号: 8553351
• 负责人: Michael J. Iadarola
• 金额: $ 69.61万
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 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/8553351
• 关键词: AIDS neuropathy; Acute; Acute Pain; Address; Affect; Animal Model; Animals; Antibodies; Antigens; Autoantibodies; Autoantigens; Autoimmune Diseases; Autoimmune Process; Autoimmune Responses; Biological; Biological Assay; Biological Markers; Body part; C Fiber; Caliber; Cancer Patient; Cell membrane; Cells; Cerebrospinal Fluid; Clinical; Clinical Research; Collaborations; Communicable Diseases; Communication; Complex; Complex Regional Pain Syndromes; Data Set; Devices; Diabetes Mellitus; Diabetic Neuropathies; Diagnostic; Disease; Elements; Evaluation; Evolution; Exhibits; Face; Facial nerve structure; Family; Fiber; Gene Expression Profile; Goals; Herpes zoster disease; Human; Human Herpesvirus 8; Human T-lymphotropic virus 1; Immune; Immune response; Immune system; Immunoprecipitation; Individual; Infection; Inflammation; Injury; Institutes; Insulin-Dependent Diabetes Mellitus; Interferon Type II; Interferons; Investigation; Ion Channel; Life; Liquid substance; Lower Extremity; Luciferases; Lyme Disease; Maintenance; Mammalian Cell; Measures; Metabolic Diseases; Methodology; Microfluidics; Modeling; Molecular; Monitor; Multiple Sclerosis; Nerve Endings; Nerve Fibers; Nervous system structure; Neurologic; Neurologic Manifestations; Neuropathy; Nociception; Non-Insulin-Dependent Diabetes Mellitus; Numbness; Operative Surgical Procedures; Opportunistic Infections; Oral cavity; Pain; Pain Disorder; Painful Paresthesias; Paper; Patients; Peripheral; Peripheral Nerves; Peripheral Nervous System Diseases; Phase; Plasma; Play; Population; Postherpetic neuralgia; Predictive Value; Process; Protein Family; Protein Fragment; Proteins; Proteome; Publishing; RNA; Recombinant DNA; Recombinant Proteins; Registries; Reporting; Research; Research Project Grants; Risk; Role; Saliva; Salivary; Sampling; School Dentistry; Sensitivity and Specificity; Sensory; Serum; Signal Pathway; Sjogren' s Syndrome; Specificity; Spinal Cord; Symptoms; Syndrome; System; TRIM Motif; Testing; Time; Tissues; Tracer; Translational Research; Trauma; Upper Extremity; Vaccines; Virus; Work; aquaporin 4; base; beta-adrenergic receptor; central nervous system injury; chemotherapeutic agent; chronic neuropathic pain; chronic pain; cytokine; drug development; human disease; insight; luminescence; member; nerve injury; nervous system disorder; painful neuropathy; patient population; point-of-care diagnostics; programs; receptor; relating to nervous system; therapeutic development; transmission process; water channel

Overview: Chronic neuropathic pain can affect any part of the body, including the oral cavity and facial nerves. Neuropathic pain can occur due to a variety of insults, infections, autoimmune disorders such as Sjogrens Syndrome, or metabolic disorders such as diabetes (diabetic neuropathy). We are testing the hypothesis that, in some patients, chronic pain is maintained by immunopathological processes related to autoantibodies generated against proteins in peripheral nerve. Autoantibodies are known culprits in certain large fiber peripheral neuropathies. 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-fiber or A-delta nerve fibers. In support of this idea, it has been reported that between 20 and 30% of Sjogrens syndrome (SjS) patients exhibit a small fiber neuropathy that produces painful paresthesias in the upper and lower extremities. Similar neuropathic pain occurs prominently in Type II diabetes and in cancer patients treated with certain chemotherapeutic agents. 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 tracers expressed in mammalian cells. This assay is called luciferase immunoprecipitation systems (LIPS) and we have published papers showing that LIPS can readily, and without interference, measure antibodies in serum, plasma, cerebrospinal fluid, saliva and other bodily fluids. This translational research program addresses molecular and pathophysiological processes of nociceptive transmission and new ways to investigate chronic pain, autoimmune conditions and infectious diseases in human patients. (1) Our main goal is to understand the molecular and cell biological mechanisms underlying human chronic pain disorders. In addition, the assay methodology we established has great versatility due to its sensitivity, modularity, and use of recombinant DNA methodology to generate protein antigen tracers. Thus, numerous conditions have been examined as we evaluate the range of diseases and disorders that antibodies play a role in. These include various infectious diseases, with and without nervous system involvement, and various autoimmune disorders, all of which have nervous system symptomology in subsets of patients. , (2) Thus a second goal is to develop new, more sensitive and robust clinical tests for relevant diseases or conditions. Last year we extended the SjS study to include a comparison of salivary antibody levels to those in serum. Using only 5 microliters, the LIPS assay readily detected the major SjS autoantigens in saliva, yielding the same sensitivity and specificity as in serum. These results highlighted the feasibility of establishing non-invasive, saliva based assays for many types of human diseases and for monitoring of vaccine immune status for large populations of people. This year we published a report on a microfluidic platform for the LIPS assay that demonstrates proof of principle for a small point-of-care diagnostic device. The major Sjogren's antigens include the protein Ro52; this is an interferon-inducible member of the tripartite motif (TRIM) family of proteins. We have discovered a new Sjogrens autoantibody from the same TRIM family and have begun to examine this more broadly. We will begin to confirm these observations using serum from the SICCA Sjogren's registry maintained at the School of Dentistry, UCSF. We also identified antibodies to interferon gamma in some of the samples and also in some of the neuropathic pain samples. We generated a larger panel of anticytokine antibody probes. Antibodies to the various cytokines were not widely represented in either population. However, anti-interferon gamma antibodies do appear to play a role opportunistic infections and this collaborative research work defines a new mechanism of disease in certain patient populations. These results were recently published. In many neural autoimmune disorders the major autoantigens are frequently plasma membrane receptors or ion channels. We have generated several such probes for neuropathic pain disorders, notably the beta-adrenergic receptor based on published reports, and are currently testing to determine if the LIPS assay can detect antibodies to this protein in appropriate individuals. We are currently working on several additional inter-institute and inter-institutional collaborations to obtain well-characterized patients with Complex Regional Pain Syndrome (CRPS, a neuropathic pain disorder), other neuropathies, and other CNS and PNS disorders and infectious diseases that have neurological manifestations, especially pain, such as shingles and post-herpetic neuralgia. One of the most compelling aspects of this project is the progressive layering and evolution of the data set. 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 a common denominator or general mechanism, and (c) antigenicity within an entire signaling pathway involved in inter- or intracellular communication. An example is the antigenicity of the TRIM family of proteins in Sjogren's Syndrome. As time progresses, full multiple antigen profiling can be implemented to obtain a new level of understanding of many complex human disease states.

概述：慢性神经性疼痛会影响身体的任何部位，包括口腔和面部神经。神经性疼痛可能是由于多种损伤，感染，自身免疫性疾病（如Sjogrens综合征）或代谢性疾病（如糖尿病（糖尿病神经病））。我们正在检验以下假设：在某些患者中，慢性疼痛通过与外周神经中蛋白质产生的自身抗体有关的免疫病理学过程维持。在某些大型纤维周围神经病中，自身抗体是已知的罪魁祸首。在疼痛是成分的地方，我们假设存在于小直径，疼痛感（伤害感受）C纤维或A-二尔特神经纤维引起的神经末端发现的蛋白质自身抗体。为了支持这一想法，据报道，20％至30％的Sjogrens综合征（SJS）患者表现出一种小的纤维神经病，在上肢和下肢会产生疼痛的异常。在II型糖尿病和用某些化学治疗剂治疗的癌症患者中，类似的神经性疼痛出现。为了测试疼痛神经性疾病具有自身免疫性成分的假设，我们建立了一种敏感，定量，液相发光测定法，该测定使用在哺乳动物细胞中表达的重组蛋白抗原示踪剂。该测定法被称为荧光素酶免疫沉淀系统（LIPS），我们发表了论文，表明嘴唇可以很容易，并且在不干预的情况下，测量血清，血浆，脑脊液，唾液，唾液，唾液和其他体液中的抗体。该转化研究计划介绍了伤害感受传播的分子和病​​理生理过程，以及研究人类患者的慢性疼痛，自身免疫性状况和感染性疾病的新方法。 （1）我们的主要目标是了解人类慢性疼痛障碍的分子和细胞生物学机制。此外，由于其灵敏度，模块化以及重组DNA方法的使用来生成蛋白质抗原示踪剂，因此我们确定的测定方法具有很大的多功能性。因此，当我们评估抗体发挥作用的疾病和疾病范围时，已经检查了许多疾病。这些疾病和疾病范围包括各种传染病，有和没有神经系统的参与以及各种自身免疫性疾病，所有这些疾病都在患者亚群中具有神经系统症状。 ，（2）因此，第二个目标是针对相关疾病或疾病开发新的，更敏感和强大的临床测试。 去年，我们扩展了SJS研究，以将唾液抗体水平与血清​​中的研究进行比较。 Lips仅使用5个微量液体，很容易检测到唾液中主要的SJS自身抗原，具有与血清相同的敏感性和特异性。这些结果强调了为许多类型的人类疾病建立非侵入性，基于唾液的测定的可行性，并监测大量人群的疫苗免疫状态。今年，我们发布了有关嘴唇测定法的微流体平台的报告，该报告证明了小型护理诊断设备的原理证明。 主要的Sjogren抗原包括蛋白质RO52；这是三方基序（Trim）蛋白质家族的干扰素诱导成员。 我们已经发现了同一装饰家族的新型自身抗体，并开始更广泛地研究了这一点。我们将开始使用在UCSF牙科学院维持的Sicca Sjogren注册表中的血清确认这些观察结果。 我们还确定了在某些样品中以及一些神经性疼痛样本中对干扰素γ的抗体。 我们生成了较大的抗细菌抗体探针。 各种细胞因子的抗体在任何一个人群中均未广泛代表。 然而，抗互化伽马抗体确实起了机会性感染的作用，而这项协作研究工作定义了某些患者人群的新机制。这些结果最近发表了。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。 我们已经针对神经性疼痛障碍产生了几种此类探针，特别是基于已发表的报告的β-肾上腺素能受体，目前正在测试以确定LIPS分析是否可以检测到适当个体中该蛋白质的抗体。我们目前正在研究其他几个跨个体和机构间合作，以获取具有复杂区域疼痛综合征（CRP，神经性疼痛障碍），其他神经病以及其他中枢神经系统和PNS疾病和PNS疾病和感染性疾病的良好特征的患者，尤其是神经系统疾病，尤其是疼痛，尤其是shingles sheringles neuralgia。 该项目最引人注目的方面之一是数据集的渐进分层和演变。随着我们增加了跨疾病和疾病的测试抗原和测定的数量，我们组装了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）涉及整个信号途径内的抗原性。一个例子是Sjogren综合征中蛋白质蛋白质家族的抗原性。随着时间的流逝，可以实施完整的多种抗原分析，以获得对许多复杂人类疾病状态的新水平。

项目成果

Recombinant expression of the AChR-alpha1 subunit for the detection of conformation-dependent epitopes in Myasthenia Gravis.

• DOI: 10.1016/j.nmd.2010.12.003
• 发表时间: 2011-03
• 期刊: NEUROMUSCULAR DISORDERS
• 影响因子: 2.8
• 作者: Ching, Kathryn H.;Burbelo, Peter D.;Kimball, Richard M.;Clawson, Lora L.;Corse, Andrea M.;Iadarola, Michael J.
• 通讯作者: Iadarola, Michael J.

Emerging tactical strategies for fighting the war on cancer based on the genetic landscape.

基于遗传景观的抗击癌症的新兴战术策略。

• 发表时间: 2011
• 期刊: American journal of translational research
• 影响因子: 2.2
• 作者: Burbelo,PeterD;Ching,KathrynH;Bren,KathleenE;Iadarola,MichaelJ
• 通讯作者: Iadarola,MichaelJ

Antibody profiling by Luciferase Immunoprecipitation Systems (LIPS).

• DOI: 10.3791/1549
• 发表时间: 2009-10-07
• 期刊: Journal of visualized experiments : JoVE
• 作者: Burbelo, Peter D;Ching, Kathryn H;Iadarola, Michael J
• 通讯作者: Iadarola, Michael J

Searching for biomarkers: humoral response profiling with luciferase immunoprecipitation systems.

• DOI: 10.1586/epr.11.23
• 发表时间: 2011-06
• 期刊: Expert review of proteomics
• 影响因子: 3.4
• 作者: Burbelo PD;Ching KH;Bren KE;Iadarola MJ
• 通讯作者: Iadarola MJ

Synthetic biology for translational research.

用于转化研究的合成生物学。

• 发表时间: 2010
• 期刊: American journal of translational research
• 影响因子: 2.2
• 作者: Burbelo,PeterD;Ching,KathrynH;Han,BrianL;Klimavicz,CaitlinM;Iadarola,MichaelJ
• 通讯作者: Iadarola,MichaelJ