Pain and Neuro-immune Signaling in S. pyogenes pathogenesis

化脓性链球菌发病机制中的疼痛和神经免疫信号传导

基本信息

批准号：
9445623
负责人：
Isaac Ming-Cheng Chiu
金额：
$ 65.61万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-25 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9445623
关键词：
Ablation Acute Acute Pain Affect Afferent Neurons Analgesics Anti-Inflammatory Agents Anti-inflammatory Bacteria Bacterial Infections Biological Assay Botulinum Toxins Brain Cell physiology Cells Chemicals Chemosensitization Clinical Complement Data Diagnosis Disease Eating Effectiveness Genetic Host Defense Human Hyperalgesia Immune Immune response Immune system Immunity Immunologics Infection Infectious Skin Diseases Inflammasome Inflammation Innate Immune Response Interleukin-1 beta Intrathecal Injections Lead Life Light Link Mechanics Mediating Microbiology Molecular Mus Natural Immunity Necrotizing fasciitis Nerve Nerve Block Nervous system structure Neurobiology Neuroimmune Neurons Neuropeptides Nociceptors Opioid Outcome Pain Pathogenesis Pathway interactions Perception Peripheral Peripheral Nerves Phagocytosis Pharmaceutical Preparations Pharyngeal structure Pharyngitis Plasmids Play Production Recruitment Activity Role Signal Pathway Signal Transduction Skin Tissue Spinal Cord Streptococcus pyogenes Streptolysins Symptoms TRPV1 gene Testing Tissues Toxin central pain experimental study feeding immunoregulation improved outcome intradermal injection killings mutant neurobehavioral neuroregulation neurotransmission neutralizing antibody neutrophil novel strategies optogenetics pathogen peripheral pain prevent relating to nervous system skills soft tissue tool

项目摘要

PROJECT SUMMARY Pain (Dolor) is one of the four cardinal signs of inflammation, and often accompanies bacterial infections. Nociceptor neurons are the peripheral sensory neurons that mediate pain, which densely innervate barrier tissues including the skin and soft tissues that are exposed to pathogens. Streptococcus pyogenes is a leading cause of necrotizing fasciitis, an invasive and life-threatening form of infection, in which pain occurs early and “out of proportion” with other symptoms. We hypothesize that pain plays a major causative role in the pathogenesis of S. pyogenes, by inducing neural mediated suppression of innate immune cell recruitment and killing of bacteria. Here, we will determine: 1) The molecular mechanisms of pain during S. pyogenes infection, with a focus on streptococcal pore-forming toxins streptolysin S (SLS) and streptolysin O (SLO), and 2) The role of nociceptors and pain signaling in regulating neutrophil function and host defense against S. pyogenes. We test the hypothesis that targeting pain signaling would lead to enhanced innate immune responses. Given the importance of pain in the diagnosis of necrotizing fasciitis, and the widespread use of analgesics, our findings connecting pain to S. pyogenes host defense could have important clinical implications. The two aims of this study leverage the complementary skills of Dr. Chiu and Dr. Wessels, combining neurobiological, immunological, and microbiological approaches to investigate the role of pain in host defense. In Specific Aim 1, we will determine the critical molecular mechanisms of pain production during S. pyogenes infection by two clinical isolates. We use isogenic mutant bacterial strains and plasmid complementation strategies together with neurobehavioral assays to determine whether SLS and SLO mediate S. pyogenes-induced pain. We will determine whether the inflammasome and IL-1β are involved in neuronal recognition of pore-forming toxins or pain signaling. In addition, we determine the effectiveness of non-steroidal anti-inflammatory drugs (NSAIDs) in impacting pain perception during S. pyogenes infection. In Specific Aim 2, we will determine how targeted ablation of TRPV1+ nociceptors using RTX treatment or Trpv1-cre/DTA mice enhances host defenses against S. pyogenes. We will determine how temporally and spatially controlled modulation of TRPV1+ neural activity using optogenetic or DREADD strategies affects the outcome of S. pyogenes infection. We will also utilize opioids to block central pain perception or Botulinum toxin to block peripheral pain signaling to determine which neural component modulates immune responses against S. pyogenes. In our analysis, we elucidate the role of nociceptor-derived neuropeptides such as CGRP in suppressing neutrophil phagocytosis and killing of S. pyogenes. This study analyzes a significant link between induction of pain signaling, neural blockade of innate immunity, and the potentiation of S. pyogenes bacterial pathogenesis. Targeting pain and neuro-immune suppression could lead to novel approaches to treatment of this and other invasive bacterial infections.

项目概要疼痛（Dolor）是炎症的四个主要症状之一，通常伴随细菌感染。伤害感受器神经元是介导疼痛的外周感觉神经元，密集地支配屏障暴露于病原体的皮肤和软组织等组织是主要的致病菌。坏死性筋膜炎的病因，这是一种侵入性且危及生命的感染形式，其中疼痛发生较早，并且我们认为疼痛与其他症状“不成比例”。化脓性链球菌的发病机制，通过诱导神经介导的先天免疫细胞募集抑制和在这里，我们将确定：1）化脓性链球菌感染期间疼痛的分子机制，重点关注链球菌成孔毒素链球菌溶血素 S (SLS) 和链球菌溶血素 O (SLO)，以及 2) 伤害感受器和疼痛信号传导在调节中性粒细胞功能和宿主对化脓性链球菌的防御中的作用。我们测试了以下假设：针对疼痛信号传导会导致先天免疫反应增强。疼痛在诊断坏死性筋膜炎中的重要性以及镇痛药的广泛使用，我们将疼痛与化脓性链球菌宿主防御联系起来的发现可能具有重要的临床意义。这项研究利用了 Chiu 博士和 Wessels 博士的互补技能，结合了神经生物学、免疫学和微生物学方法研究疼痛在宿主防御中的作用。 1，我们将通过两种方法确定化脓性链球菌感染期间疼痛产生的关键分子机制我们同时使用同基因突变菌株和质粒互补策略。通过神经行为测定来确定 SLS 和 SLO 是否介导化脓性链球菌引起的疼痛。确定炎症小体和 IL-1β 是否参与神经元对成孔毒素的识别或此外，我们还确定了非甾体抗炎药 (NSAID) 在治疗中的有效性。在具体目标 2 中，我们将确定如何针对性地影响化脓性链球菌感染期间的疼痛感知。使用 RTX 治疗或 Trpv1-cre/DTA 小鼠消除 TRPV1+ 伤害感受器可增强宿主对伤害感受器的防御我们将确定如何在时间和空间上控制 TRPV1+ 神经活动的调节。使用光遗传学或 DREADD 策略会影响化脓性链球菌感染的结果。阿片类药物阻断中枢疼痛感知或肉毒杆菌毒素阻断外周疼痛信号传导以确定哪种药物神经成分调节针对化脓性链球菌的免疫反应在我们的分析中，我们阐明了其作用。伤害感受器衍生的神经肽，例如 CGRP，可抑制中性粒细胞吞噬作用并杀死金黄色葡萄球菌。这项研究总结了疼痛信号传导与先天神经阻断之间的重要联系。免疫，以及化脓性链球菌细菌发病机制的增强，针对疼痛和神经免疫。抑制可能会导致治疗这种和其他侵袭性细菌感染的新方法。