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.

慢性神经性疼痛可以影响身体的任何部位，包括口腔和面部神经。神经性疼痛可由于多种损伤、感染、自身免疫性疾病（例如舍格伦综合征）或代谢性疾病（例如糖尿病（糖尿病性神经病））而发生。我们正在测试的假设，在一些患者中，慢性疼痛是由免疫病理过程相关的自身抗体产生的周围神经蛋白质。自身抗体是某些大纤维周围神经病变的罪魁祸首。在疼痛是一个组成部分，我们假设存在的自身抗体的蛋白质中发现的神经末梢所产生的小直径，疼痛感（伤害性）C-纤维或A-δ神经纤维。为了支持这一观点，据报道，20%至30%的干燥综合征（SjS）患者表现出小纤维神经病变，在上肢和下肢产生疼痛性感觉异常。类似的神经性疼痛主要发生在II型糖尿病和用某些化疗剂治疗的癌症患者中。为了检验疼痛性神经病性病症具有自身免疫组分的假设，我们建立了一种灵敏的、定量的、液相发光测定法，该测定法使用在哺乳动物细胞中表达的重组蛋白抗原示踪剂。这种检测被称为荧光素酶免疫沉淀系统（LIPS），我们已经发表的论文表明，LIPS可以很容易地，没有干扰，测量血清，血浆，脑脊液，唾液和其他体液中的抗体。这个翻译研究计划涉及伤害性传递的分子和病理生理过程，以及研究人类患者慢性疼痛，自身免疫性疾病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，我们建立的检测方法具有很大的通用性，由于其灵敏度，模块化，并使用重组DNA方法来产生蛋白质抗原示踪剂。因此，在我们评估抗体发挥作用的疾病和病症的范围时，已经检查了许多条件。这些包括各种感染性疾病，有和没有神经系统的参与，和各种自身免疫性疾病，所有这些都有神经系统病理学的患者子集。因此，第二个目标是开发用于相关疾病或病症的新的、更灵敏和更稳健的临床测试。 去年，我们扩展了SjS研究，将唾液抗体水平与血清抗体水平进行了比较。仅使用5微升，LIPS检测很容易检测唾液中的主要SjS自身抗原，产生与血清相同的灵敏度和特异性。这些结果强调了建立用于许多类型的人类疾病的非侵入性的基于唾液的测定和用于监测大人群的疫苗免疫状态的可行性。今年，我们发表了一份关于LIPS检测的微流体平台的报告，该报告证明了小型即时诊断设备的原理。 主要的干燥抗原包括Ro 52蛋白;这是干扰素可诱导的TRIM蛋白家族成员。 我们发现了一种来自同一TRIM家族的新的Sjogrens自身抗体，并开始更广泛地研究这一点。我们将开始使用来自加州大学旧金山分校牙科学院SICCA Sjogren登记处的血清来证实这些观察结果。 我们还在一些样本和一些神经性疼痛样本中鉴定了干扰素γ的抗体。 我们产生了一个更大的抗细胞因子抗体探针组。 各种细胞因子的抗体在两个人群中都没有广泛代表。 然而，抗干扰素γ抗体似乎确实在机会性感染中发挥作用，这项合作研究工作在某些患者群体中定义了一种新的疾病机制。这些结果最近发表。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。 我们已经产生了几个这样的探针的神经性疼痛疾病，特别是β-肾上腺素能受体的基础上发表的报告，目前正在测试，以确定是否LIPS检测可以检测抗体，这种蛋白质在适当的个人。我们目前正在进行几个额外的机构间和机构间合作，以获得具有复杂区域疼痛综合征（CRPS，一种神经性疼痛疾病），其他神经病，以及其他CNS和PNS疾病和具有神经学表现的感染性疾病的良好特征的患者，特别是疼痛，如带状疱疹和疱疹后神经痛。 该项目最引人注目的方面之一是数据集的逐步分层和演变。随着我们增加测试抗原的数量和检测条件和疾病，我们组装免疫和自身免疫反应的综合评估。这是通过确定（a）对正向同源蛋白和蛋白片段的免疫应答的程度和特异性，（B）指示共同特征或一般机制的抗原谱的重叠，和（c）参与胞间或胞内通信的整个信号传导途径内的抗原性来实现的。一个例子是干燥综合征中TRIM蛋白家族的抗原性。随着时间的推移，可以实施完整的多抗原分析，以获得对许多复杂人类疾病状态的新水平的理解。

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.

搜索生物标志物：使用荧光素酶免疫沉淀系统进行体液反应分析。

• 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