ILC2 memory in asthma

哮喘中的 ILC2 记忆

基本信息

批准号：
10267732
负责人：
Rafeul Alam
金额：
$ 62.14万
依托单位：
NATIONAL JEWISH HEALTH
依托单位国家：
美国
项目类别：
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/10267732
关键词：
ATAC-seq Abbreviations Ablation Adoptive Transfer Adult Adult asthma Allergens Allergic Alternaria Antigens Asthma B-Lymphocytes BACH2 gene Cell model Chronic Chronic Obstructive Airway Disease Clinical Cytokine Gene Dose Environmental Risk Factor Eosinophilia 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 Receptor Signaling Refractory Reporting Repression Research Role Saline Sampling 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 asthmatic patient base cross reactivity dectin 1 disorder control eosinophilic asthma eosinophilic inflammation experimental study gene repression inhibitor/antagonist 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

项目摘要

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人哮喘人群的 30%，通常与以下疾病相关：与过敏性哮喘不同，非过敏性哮喘的机制尚不清楚。嗜酸性粒细胞炎症在非过敏性哮喘中很常见，环境因素是否会导致这种情况。嗜酸性粒细胞炎症的发病机制尚不清楚。我们通过建立哮喘小鼠模型。重复暴露于过敏原相关的 PAMP（病原体相关分子模式）。淋巴样细胞）从这个模型中发展出一种 PAMP 特异性记忆，可以通过 3-15周后，根据广泛的初步结果，我们陷入了困境。重复接触 PAMPS 会导致 ILC 记忆的形成，它由两个程序组成：基因抑制计划和准备计划对先前激活的基因进行抑制。记忆的遗传基础。这种抑制可以防止宿主在缺乏 PAMP 的情况下受到损害。准备计划通过激活 Fhl2-ERK1/2-AP1 通路，解除先前标记基因的抑制并诱发哮喘（图 1）。测试这一假设提议的 3 个具体目标在目标 1 下，我们将研究过敏原和 PAMP 引发的哮喘。和小鼠记忆 ILC，并检查过敏原和 PAMP 之间的交叉反应性。在目标 2 下，我们将阐明 ILC 和模式识别受体在该哮喘模型中的重要性。我们将研究 ILC 记忆的形成机制。过敏原和 PAMP 诱导的记忆 ILC 及其随时间的持久性。来自记忆 ILC 的 ATAC-seq 表明阻遏物 Bach2 和准备途径 Fhl2- 的作用 ERK1/2-Fosb 分别对记忆诱导和回忆起作用。我们将研究 Fhl2、Bach2 和 FosB 的作用。目标 3 将致力于人类 ILC 的研究。人类 ILC 中的抑制和准备程序基因的研究，并研究其诱导机制。我们将研究嗜酸性非嗜酸性粒细胞中记忆 ILC 的频率及其对过敏原和 PAMP 的反应。我们将采用公正且稳健的基因组来研究过敏性哮喘患者并建立其临床相关性。 RNA-seq 和 ATAC-seq 等方法，并利用创新的小鼠模型来测试这一新概念 PAMP 引发的 ILC 记忆我们将研究非过敏性嗜酸性粒细胞性哮喘的分子机制。测试治疗策略，并寻求使用哮喘患者的临床样本建立人类相关性我们拥有必要的专业知识和工具来进行拟议的实验。产生范式转变的新知识和干预策略，有助于治疗非过敏性哮喘。