Functional role of miR-511-3p in allergic asthma and its underlying mechanisms

miR-511-3p在过敏性哮喘中的功能作用及其潜在机制

基本信息

批准号：
10385822
负责人：
Peisong Gao
金额：
$ 62.31万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-07 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10385822
关键词：
Affinity Allergens Allergic Allergic inflammation Anti-Inflammatory Agents Antigens Asthma Binding Biotin Clustered Regularly Interspaced Short Palindromic Repeats Cost Measures Critical Care Data Set Defense Mechanisms Dependovirus Development Dictyoptera Economic Burden Environmental Exposure Equilibrium Exposure to Extrinsic asthma Genes Genetic Transcription Genome engineering Goals Human Hypersensitivity Immune response Inflammatory Inhalation Knock-out Knockout Mice Lead Link Longitudinal Studies Luciferases Lung Mannose Mediating Medicine Messenger RNA MicroRNAs Molecular Mus Patients Phenotype Pilot Projects Plasma Polysaccharides Prevalence Prostaglandin D2 Public Health Reporter Research Risk Role Sampling Severities Signal Pathway Signal Transduction Sputum TLR4 gene Testing Transcript Untranslated RNA airway hyperresponsiveness airway inflammation allergic airway inflammation asthma exacerbation asthma model asthmatic base cockroach allergen conditional knockout environmental allergen experimental study extracellular vesicles human subject innovation macrophage mannose receptor mouse model new therapeutic target novel novel diagnostics overexpression peripheral blood prophylactic tool transcriptomics

项目摘要

ABSTRACT Exposure to cockroach allergen can lead to allergic sensitization and an increased risk of allergic asthma. However, the underlying molecular mechanisms are currently not well-established. Our long-term goals are to elucidate the fundamental underlying mechanisms and identify novel therapeutic targets for allergic asthma. During the pilot studies, our group has made significant contributions to unraveling an important link between cockroach antigen and development of allergic asthma. Specifically, our profiling of N-linked glycans from cockroach allergen identified several major glycans with high affinity to mannose receptor, MRC1/CD206. Furthermore, we have identified a critical but previously unrecognized role of MRC1 in allergen clearance as a natural defense mechanism and in limiting the progression and severity of cockroach allergen-induced allergic inflammation in a mouse model of asthma. This occurs through alterations in macrophage clearance of the inhaled cockroach allergens and balance of M1/M2 macrophage polarization. This was at first perplexing because MRC1 lacks any known signaling motif, therefore, the signaling cascades of MRC1 in allergen-induced airway inflammation and macrophage polarization remain obscure. Our breakthrough for a deeper understanding of the MRC1 signaling pathway came with the recognition that a key regulatory miR-511-3p, encoded by both mouse and human MRC1 gene, is transcriptionally co-regulated with MRC1 in macrophages. These exciting findings lead us to propose a novel hypothesis that MRC1 is largely involved in allergen clearance as a natural defense mechanism, and MRC1-encoded miR-511-3p is involved in mediating MRC1 downstream immune responses and protecting against allergen-induced airway inflammation. This hypothesis is further buttressed by our recent findings that plasma levels of miR-511-3p were much lower in asthmatics compared to controls, and that adeno-associated virus (AAV)-mediated miR-511-3p over-expression ameliorated the allergen-induced airway inflammation in Mrc1-/- mice, but miR-511-3p knockout mice showed increased allergen-induced airway inflammation. These exciting data set the stage to critically evaluate the functional significance of miR-511-3p in allergic asthma and its underlying mechanisms. Three independent yet related specific aims are proposed. Aim 1 will determine the significance of miR-511-3p in allergic asthma by quantifying miR-511-3 in plasma, sputum and extracellular vesicles (EVs) from plasma and sputum of allergic asthmatics and testing its role in macrophage polarization and function. Aim 2 will define whether miR-511-3p protects against asthma using miR-511-3p global and macrophage conditional knockout mice (e.g., LysM-cre; miR-511-3pflox/flox) and mannose-decorated EV-miR- 511-3p-treated mice. Aim 3 will identify miR-511-3p targets by integrating gene profiling and our unique affinity- based transcriptomic approach for miR-511-3p binding partner mRNAs/long non-coding RNAs. Collectively, our studies will provide a conceptual framework linking allergens, MRC1, and miR-511-3p to key features of asthma. Ultimately, these studies may allow for the development of new diagnostic and therapeutic targets for asthma.

抽象的暴露于蟑螂过敏原会导致过敏反应和过敏性哮喘的风险增加。但是，目前尚不确定的基本分子机制。我们的长期目标是阐明基本的基本机制，并确定过敏性哮喘的新型治疗靶标。在试点研究期间，我们的小组为解开之间的重要联系做出了重大贡献蟑螂抗原和过敏性哮喘的发育。具体而言，我们从蟑螂过敏原确定了几种对甘露糖受体MRC1/CD206高亲和力的主要聚糖。此外，我们已经确定了MRC1在过敏原清除中的关键但以前未认识到的作用自然防御机制，并限制蟑螂过敏性过敏的进展和严重程度哮喘小鼠模型中的炎症。这是通过改变巨噬细胞清除的吸入蟑螂过敏原和M1/M2巨噬细胞极化的平衡。一开始很困惑由于MRC1缺少任何已知的信号基序，因此，过敏原诱导的MRC1的信号级联气道炎症和巨噬细胞极化仍然晦涩。我们深入了解的突破 MRC1信号传导途径的认识是，密钥监管mir-511-3p，两者都编码小鼠和人类MRC1基因在巨噬细胞中与MRC1共同调节。这些令人兴奋调查结果使我们提出了一个新的假设，即MRC1在很大程度上参与过敏原清除为自然防御机制和MRC1编码的miR-511-3p参与介导MRC1下游免疫反应并防止过敏原引起的气道炎症。这个假设进一步支持我们最近的发现，与对照组相比，哮喘患者的血浆水平低得多，并且与腺体相关病毒（AAV）介导的miR-511-3p过表达改善了过敏原诱导的 MRC1 - / - 小鼠的气道炎症，但miR-511-3p敲除小鼠显示过敏原诱导的气道增加炎。这些令人兴奋的数据设定了批判性评估miR-511-3p的功能意义的阶段过敏性哮喘及其基本机制。提出了三个独立但相关的特定目标。目的 1将通过量化血浆中的miR-511-3，确定miR-511-3p在过敏性哮喘中的重要性和过敏性哮喘的血浆和痰液中的细胞外囊泡（EV），并测试其在巨噬细胞中的作用极化和功能。 AIM 2将使用miR-511-3p Global定义miR-511-3p是否保护哮喘和巨噬细胞条件敲除小鼠（例如Lysm-cre； mir-511-3pflox/flox）和甘露糖（Mannose）装饰的EV-MIR- 511-3p处理的小鼠。 AIM 3将通过整合基因分析和我们独特的亲和力 - miR-511-3p结合伴侣mRNA/长非编码RNA的基于基于的转录组方法。总体而言，我们的研究将提供一个概念框架，将过敏原，MRC1和miR-511-3p与哮喘的关键特征联系起来。最终，这些研究可能允许开发哮喘的新诊断和治疗靶标。