ILC2 memory in asthma

哮喘中的 ILC2 记忆

• 批准号: 10685256
• 负责人: Rafeul Alam
• 金额: $ 60.64万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-21 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10685256
• 关键词: ATAC-seq; Abbreviations; Ablation; Adoptive Transfer; Adult; Adult asthma; Allergens; Allergic; Alternaria; Antigens; Asthma; Asthma COPD Overlap Syndrome; Atopic Dermatitis; B-Lymphocytes; BACH2 gene; Chronic; Clinical; Cytokine Gene; Dose; Environmental Risk Factor; Eosinophilia; Eosinophilic Esophagitis; Epigenetic Process; Exposure to; Extrinsic asthma; Family; Flow Cytometry; Frequencies; Genes; Genetic; Genomic approach; Human; IL5 gene; Immune; Intervention; Knowledge; LIM Domain; Lymphoid Cell; MAPK3 gene; Memory; Modeling; Molecular; Mus; Oncogenes; Pathogenesis; Pathway interactions; Patients; Pattern; Pattern recognition receptor; Pharmaceutical Preparations; Play; Population; Production; Property; Proteinase-Activated Receptors; Rag1 Mouse; Readiness; Refractory; Reporting; Repression; Research; Role; Saline; Sampling; Signal Transduction; Source; Stains; Steroid Resistance; Steroids; Subtilisins; T-Lymphocyte; Testing; Th2 Cells; Therapeutic; Time; Transcription Factor AP-1; Western Blotting; Wild Type Mouse; airway hyperresponsiveness; asthma model; asthmatic patient; cross reactivity; dectin 1; derepression; disorder control; eosinophilic asthma; eosinophilic inflammation; experimental study; gene repression; inhibitor; innovation; loss of function; mouse model; new therapeutic target; novel; osteosarcoma; pathogen; prevent; programs; receptor; reconstitution; response; single-cell RNA sequencing; therapeutic evaluation; tool; transcriptome sequencing; transcriptomics

ABSTRACT Non-allergic asthma constitutes about 30% of the adult asthma population and is frequently associated with severe asthma. Unlike that of allergic asthma the mechanism of non-allergic asthma is poorly understood. Eosinophilic inflammation is common in non-allergic asthma. Whether environmental factors contribute to the pathogenesis of eosinophilic inflammation is unclear. We have developed a mouse model of asthma through repetitive exposure to allergen-associated PAMPs (Pathogen-associated Molecular Patterns). ILCs (innate lymphoid cells) from this model developed a PAMP-specific memory, which could be elicited with a subthreshold dose of the PAMP 3-15 weeks later. Based upon extensive preliminary results, we hypothesize that repetitive exposure to PAMPS leads to the formation of ILC memory, which constitutes two programs—a gene repression program and a preparedness program. The repression of previously activated genes forms the genetic basis for memory. This repression prevents host damage in the absence of the PAMPs. The preparedness program generates a rapid response to a subthreshold PAMP exposure through activation of the Fhl2-ERK1/2-AP1 pathway, which de-represses the previously marked genes and induces asthma (Fig. 1). We propose 3 specific aims to test this hypothesis. Under Aim 1 we will study allergen- and PAMP-elicited asthma and memory ILCs in mice and examine cross-reactivity among allergens and PAMPs. We will assess the importance of ILCs and the pattern recognition receptors in this asthma model. Under aim 2 we will elucidate the mechanism of formation of ILC memory. We will study the transcriptomic and epigenetic landscapes of allergen- and PAMP-induced memory ILCs and their persistence over time. The results from scRNA-seq and ATAC-seq from memory ILCs suggested a role for the repressor Bach2 and the preparedness pathway Fhl2- ERK1/2-Fosb for memory induction and recall, respectively. We will examine the role of Fhl2, Bach2 and FosB in a loss-of-function approach. Aim 3 will be devoted to human ILCs studies. We will examine the expression of the repression and preparedness program genes in human ILCs and study the mechanism of their induction. We will study the frequency of memory ILCs and their response to allergens and PAMPs in eosinophilic non- allergic asthmatic patients and establish their clinical correlation. We will employ unbiased and robust genomic approaches such as RNA-seq and ATAC-seq, and utilize innovative mouse models to test the novel concept of PAMP-elicited ILC memory. We will examine the molecular mechanism of non-allergic eosinophilic asthma, test therapeutic strategies, and seek to establish human relevance using clinical samples from asthmatic patients. We have the necessary expertise and tools to conduct the proposed experiments. The proposal will generate paradigm-shifting new knowledge and intervention strategies that will help treat non-allergic asthma.

抽象的 非过敏性哮喘约占成年哮喘的30％，并且经常与 严重的哮喘。与过敏性哮喘不同，非过敏性哮喘的机制知之甚少。 嗜酸性注射在非过敏性哮喘中很常见。环境因素是否有助于 嗜酸性炎症的发病机理尚不清楚。我们已经通过 重复暴露于与过敏原相关的PAMP（病原体相关的分子模式）。 ILCS（先天 该模型中的淋巴细胞）开发了一种PAMP特异性记忆，可以用A 3-15周后，幼虫的亚阈值剂量。基于广泛的初步结果，我们假设 重复接触PAMPS导致ILC内存的形成，这构成了两个程序 - A 基因表达程序和准备计划。先前激活的基因的表达 记忆的遗传基础。这种表达在没有弹药的情况下阻止了宿主损害。 准备计划通过激活来产生对亚阈值PAMP暴露的快速响应 FHL2-ERK1/2-AP1途径，它驱除了先前标记的基因并诱导哮喘（图1）。我们 提案3的特定旨在检验这一假设。在AIM 1下，我们将研究过敏原和PAMP引发的哮喘 小鼠中的记忆ILC并检查过敏原和弹药的交叉反应性。我们将评估 在此哮喘模型中，ILC和模式识别受体的重要性。在AIM 2下，我们将阐明 ILC记忆形成的机理。我们将研究的转录组和表观遗传景观 过敏原和弹药诱导的记忆ILC及其持久性随着时间的流逝。 Scrna-Seq和 来自记忆ILC的ATAC-SEQ提出了复制品Bach2的作用和准备途径FHL2-- ERK1/2-FOSB分别用于记忆诱导和回忆。我们将研究FHL2，Bach2和FOSB的作用 在功能丧失的方法中。 AIM 3将专门研究人类ILCS研究。我们将检查表达 人类ILC中的表达和制备程序基因，并研究其诱导的机制。 我们将研究记忆ILC的频率及其对嗜酸性非嗜酸性过敏原和症状的反应 过敏性哮喘患者并建立其临床相关性。我们将采用公正和强大的基因组 RNA-Seq和Atac-Seq等方法，并利用创新的鼠标模型来测试新颖的概念 PAMP引用的ILC内存。我们将检查非过敏性嗜酸性哮喘的分子机制， 测试治疗策略，并寻求使用哮喘的临床样本建立人类相关性 患者。我们有必要的专业知识和工具来进行拟议的实验。提案将 产生范式转移新知识和干预策略，以帮助治疗非过敏性哮喘。