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

• 批准号: 9569582
• 负责人: Isaac Ming-Cheng Chiu
• 金额: $ 63.87万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 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/9569582
• 关键词: Ablation; Acute; Acute Pain; Affect; Afferent Neurons; Analgesics; Bacteria; Bacterial Infections; Biological Assay; Botulinum Toxins; Brain; Cell physiology; Cells; Chemicals; Chemosensitization; Clinical; Complement; Data; Diagnosis; Disease; Eating; Effectiveness; Exposure to; 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; Non-Steroidal Anti-Inflammatory Agents; Opioid; Outcome; Pain; Pathogenesis; Pathway interactions; Peripheral; Peripheral Nerves; Phagocytosis; Pharyngeal structure; Pharyngitis; Plasmids; Play; Production; Role; Signal Pathway; Signal Transduction; Skin Tissue; Spinal Cord; Streptococcus pyogenes; Streptolysins; Symptoms; TRPV1 gene; Testing; Tissues; Toxin; central pain; designer receptors exclusively activated by designer drugs; experimental study; feeding; immunoregulation; improved outcome; intradermal injection; mutant; neurobehavioral; neuroregulation; neurotransmission; neutralizing antibody; neutrophil; novel strategies; opioid injection; optogenetics; pain perception; pain signal; pathogen; peripheral pain; prevent; recruit; relating to nervous system; skills; soft tissue; tool

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.

项目摘要 疼痛（Dolor）是感染的四个基本迹象之一，通常涉及细菌感染。 伤害感受器神经元是介导疼痛的周围感官神经元，应支配屏障 包括皮肤和暴露于病原体的软组织在内的组织。链球菌为链球菌是领先的 坏死性筋膜炎的原因，一种侵入性和威胁生命的感染形式，疼痛发生早期发生 与其他症状的“不成比例”。我们假设疼痛在 链霉菌的发病机理，通过诱导神经介导的先天免疫细胞募集和 杀死细菌。在这里，我们将确定：1）链球菌感染期间疼痛的分子机制， 专注于链球菌孔形成毒素链霉素S（SLS）和链霉菌素O（SLO），以及2） 伤害感受器和疼痛信号传导在确定中性粒细胞功能和宿主防御化链球菌中的作用。 我们测试了靶向疼痛信号传导将导致先天免疫反应增强的假设。给出 疼痛在坏死性筋膜炎诊断中的重要性以及镇痛药的宽度使用，我们 将疼痛与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。两个目标 这项研究利用了Chiu博士和Wessels博士的完整技能，结合了神经生物学， 免疫学和微生物方法研究疼痛在宿主防御中的作用。在特定目标中 1，我们将通过两个 临床分离株。我们一起使用等源性突变细菌菌株和质粒完成策略 使用神经行为测定法，以确定SLS和SLO是否介导了化脓性链球菌诱导的疼痛。我们将 确定炎性体和IL-1β是否参与神经元识别孔形成毒素或 疼痛信号传导。此外，我们确定了非甾体类抗炎药（NSAID）在 在链球菌感染期间影响疼痛感。在特定目标2中，我们将确定目标 使用RTX治疗或TRPV1-CRE/DTA小鼠消融TRPV1+伤害感受器可增强宿主防御能力 链球菌。我们将确定TRPV1+神经活动的暂时和空间控制调制 使用光遗传学或DREADD策略会影响链球菌感染的结果。我们还将利用 阻塞中心疼痛感知或肉毒杆菌毒素以阻止外周疼痛信号传导以确定哪个是哪个 神经成分调节针对链球菌的免疫反应。在我们的分析中，我们阐明了 伤害感受器衍生的神经肽，例如CGRP抑制神经磷酸化和杀死S. 地源。这项研究分析了疼痛信号传导的诱导，先天性封锁之间的重要联系 免疫力和链球菌细菌发病机理的潜力。靶向疼痛和神经免疫 抑制可能导致对这种和其他侵入性细菌感染的新方法。