Unraveling transcriptomic and functional changes to immune sensing neuronal ganglia in response to allergy and bacteria

揭示免疫传感神经元响应过敏和细菌的转录组和功能变化

• 批准号: 10617638
• 负责人: Nicholas Jendzjowsky
• 金额: $ 19.56万
• 依托单位: LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-06 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10617638
• 关键词: Ablation; Affect; Allergens; Allergic; Allergic Disease; Alternaria; American; Anti-Inflammatory Agents; Antibodies; Asthma; Bacteria; Bacterial Infections; Bioinformatics; Bronchoalveolar Lavage Fluid; C Fiber; C57BL/6 Mouse; Carotid Body; Cells; Chemoreceptors; Coupled; Data; Development; Diphtheria Toxin; Disease; Electrophysiology (science); Eosinophilic Infiltrate; Epithelium; Female; Flow Cytometry; Future; Ganglia; Gender; Gene Expression; Goals; Gonadal Steroid Hormones; Harvest; Health; Host Defense; Hypersensitivity; IgE; Immune; Immune response; Immune signaling; Immunity; Immunoglobulin G; Immunoglobulins; Immunologic Surveillance; Inflammation; Inflammatory; Inhalation; Injections; International; Investigation; Ion Channel; Knowledge; Lung; Lung immune response; Methods; Microinjections; Modeling; Molecular; Mus; Nerve; Nervous System; Neuroendocrine Cell; Neuroimmune; Neurons; Neurosciences; Nodose Ganglion; Peripheral; Phenotype; Phospholipids; Play; Pneumonia; Population; Process; Qualifying; Receptor Gene; Reflex action; Regulation; Resources; Role; Saline; Sensory Ganglia; Sepsis; Sex Differences; Signal Transduction; Specific qualifier value; Stimulus; Streptococcal Infections; Streptococcus pneumoniae; T cell infiltration; TRPV1 gene; Techniques; Technology; Testing; Transgenic Mice; Tyrosine 3-Monooxygenase; Universities; Vanilloid; cell type; cytokine; diphtheria toxin receptor; immune function; immunoglobulin receptor; immunomodulatory therapies; immunoregulation; in vivo; intravital imaging; knock-down; loss of function; lung histology; male; multidisciplinary; neural; neutrophil; new therapeutic target; receptor; receptor expression; respiratory; response; sensor; sex; single-cell RNA sequencing; targeted treatment; tissue preparation; transcriptome; transcriptomics; γδ T cells

Project Summary Allergies affect millions of Americans and are exacerbated by bacterial infection. Despite mainstay therapies that suppress immune responses to allergy or bacteria, these health issues continue to persist. An unexplored target for new therapies lies with neural surveillance and regulation of immunity. However, this neural influence is poorly understood and seemingly paradoxical: immune responses are exacerbated in allergy, but in response to bacterial infection, the neural response reduces inflammation. How might this happen? One possibility is that the peripheral neural-immune sensing apparatus might be altered by such factors as immunoglobulin profiles, and gender, implying the participation of sex hormones. We hypothesize that such variables affect the transcriptome and thereby the excitability of the vagus and carotid bodies. Studies in AIM 1 will use single-cell RNAseq analysis of vagal and carotid body ganglia from male and female mice with asthma, pneumonia to identify how these nerves are changed in response to each condition. Then, we will assess nerve activity in response to immunoglobulin stimuli of each ganglia using state-of-the-art electrophysiological recording techniques. This project will reveal which neural cluster (vagus vs. carotid body) is the predominant sensory ganglia involved in regulating allergic vs. bacterial immune responses and how sex may affect neuro- immune signalling. Studies in AIM 2 will utilize DTR (diphtheria toxin receptor) technology in mice to selectively ablate cells where DTR expression is driven by TRPV1 (transient receptor potential vanilloid 1) or TH (tyrosine hydroxylase). Use of these transgenic mice will allow ablation of either vagal (TRPV1-DTR) or carotid body (TH- DTR) centers by direct neural microinjection of diphtheria toxin (DTX) into vagal or carotid body ganglia using saline injections as control. We will then expose mice to either allergen or bacterial infections. DTX injections will selectively abolish neuronal function and reveal which ganglion is the predominant neuro-immune sensor in the specified condition in male and female mice. These data will: 1) identify how the response to allergic or bacterial stimuli alters receptor gene expression in order to identify the phenotypic switch from anti- to pro-inflammatory neural-immune signalling; 2) establish a model and sex differences that will set the stage for further investigation involving targeted knockdown within neuro-immune sensing ganglia to discern function; 3) validate our models and methods for future investigations into the role of neural-immune sensing in disease states.

项目摘要 过敏症影响着数百万美国人，并因细菌感染而加剧。尽管主流疗法， 尽管抑制了对过敏或细菌的免疫反应，但这些健康问题仍然存在。一个未探测的目标 新疗法的关键在于神经监视和免疫调节。然而，这种神经影响是 人们对此知之甚少，似乎自相矛盾：免疫反应在过敏中加剧，但在对过敏反应的反应中， 细菌感染时，神经反应减少炎症。怎么会这样呢？ 一种可能性是，外周神经免疫感应装置可能被诸如以下因素改变： 免疫球蛋白谱和性别，暗示性激素的参与。我们假设， 变量影响转录组，从而影响迷走神经和颈动脉体的兴奋性。AIM 1研究 将使用来自患有哮喘的雄性和雌性小鼠的迷走神经和颈动脉体神经节的单细胞RNAseq分析， 以确定这些神经如何响应每种情况而变化。然后，我们将评估神经 使用最先进的电生理学技术， 记录技术。这个项目将揭示哪一个神经簇（迷走神经与颈动脉体）是主要的 感觉神经节参与调节过敏与细菌免疫反应，以及性别如何影响神经元 免疫信号AIM 2的研究将利用DTR（白喉毒素受体）技术在小鼠中选择性地 消融DTR表达由TRPV 1（瞬时受体电位香草酸1）或TH（酪氨酸）驱动的细胞 羟化酶）。这些转基因小鼠的使用将允许迷走神经（TRPV 1-DTR）或颈动脉体（TH-1）的消融。 将白喉毒素（DTX）直接神经显微注射到迷走神经节或颈动脉体神经节， 盐水注射作为对照。然后，我们将小鼠暴露于过敏原或细菌感染。DTX注射将 选择性地消除神经元功能，并揭示哪一个神经节是主要的神经免疫传感器。 雄性和雌性小鼠中的特定条件。 这些数据将：1）确定对过敏或细菌刺激的反应如何改变受体基因表达， 为了确定从抗炎到促炎神经免疫信号传导的表型转换; 2）建立一个 模型和性别差异，这将为进一步研究奠定基础， 神经免疫感觉神经节识别功能; 3）验证我们的模型和方法，为未来的研究 神经免疫感应在疾病状态中的作用。