Mechanism of CX3CR1+ macrophage-mediated resolution of eosinophilic allergic lung inflammation

CX3CR1巨噬细胞介导的嗜酸性过敏性肺部炎症消退机制

• 批准号: 10210655
• 负责人: Gye Young Park
• 金额: $ 58.56万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-10 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10210655
• 关键词: Active Sites; Adopted; Allergens; Allergic; Allergic inflammation; Alveolar Macrophages; Apoptotic; Asthma; Binding; Biological Assay; Bronchial Provocation Tests; CCL26 gene; CX3CL1 gene; Cell Death; Cell physiology; Cells; Characteristics; Complement 1q; Cytometry; Data; Eosinophilia; Excision; Extrinsic asthma; Family; Functional disorder; Gene Expression Profile; Gene Expression Profiling; Healthcare; Homeostasis; Human; In Vitro; Inflammation; Inflammatory; Institutional Review Boards; Investigation; Lead; Ligands; Liquid substance; Longevity; Lung; Lung Inflammation; Macrophage Activation; Mediating; Modeling; Molecular; Morphology; Mus; Organ; Pathogenesis; Pathway interactions; Patients; Phagocytes; Phase; Phenotype; Play; Prevention; Process; Protocols documentation; Refractory; Reporter; Research; Resolution; Role; Series; Site; Societies; Source; Structure of parenchyma of lung; Techniques; Testing; Time; Tissues; Transcript; Transgenic Mice; airway inflammation; asthma model; asthmatic; base; care burden; cell type; chemokine; conditional knockout; eosinophil; experimental study; gain of function; improved; in vivo; in vivo Model; inflammatory lung disease; insight; macrophage; mouse model; novel; pulmonary function; receptor; recruit; response; sialic acid binding Ig-like lectin; single-cell RNA sequencing; targeted treatment; transcriptome sequencing; translational study

Title: Mechanism of CX3CR1+ macrophage-mediated resolution of eosinophilic allergic lung inflammation Abstract: Recent studies show that tissue-resident macrophages participate in not only the initiation of inflammation but also in the resolution and prevention of local inflammation. To precisely determine the subsets of macrophages engaged in resolving lung inflammation and gain insight into their functions, we adopted new techniques of mass cytometry and single-cell RNA-seq (sc-RNA-seq) to analyze human and mouse macrophages in the lung. Our supporting data showed that alveolar macrophages (AMs) are phenotypically diverse and highly dynamic in response to allergen challenge. Based on the sc-RNA-seq data, AMs can be clustered into a few groups at a steady status. Among these groups, a subset of CX3CR1-expressing AMs (CX3CR1+ AMs) are unique in terms of their phenotype and patterns of gene expression, compared to classical resident AMs which are CX3CR1 negative. In patients with allergic asthma and a mouse model of asthma, we found that CX3CR1+ AMs are markedly increased in BAL by allergen challenge. The CX3CR1+ AMs express not only the macrophage but also eosinophil markers such as human Siglec-8. Further investigation with the CX3CR1- reporter and Epx-cre (a.k.a. Eo-cre) reporter mice reveals that CX3CR1+ macrophages engulf eosinophils at a steady state and in allergic lung inflammation. Depletion of CX3CR1+ macrophages in mouse models resulted in spontaneous tissue eosinophilia at a steady status and prolonged tissue eosinophilia in allergic lung inflammation. Based on this data, we hypothesized that the newly recruited CX3CR1+ AM subset promote the clearance of tissue eosinophils and facilitates the resolution of allergic lung inflammation. In aim 1, we will focus on the cellular dynamics of CX3CR1+ macrophages in allergic lung inflammation. Regarding the molecular mechanism of CX3CR1+ mediated-eosinophil clearance, we examined the potential ligands for CX3CR1 – CX3CL1 and CCL26. We discovered that CCL26 plays a key role in activating CX3CR1+ macrophages, whereas CX3CL1 is indispensable. Our sc-RNA-seq data revealed that CX3CR1+ AM subset is the sole source of the transcript of C1q - a key molecule for efferocytosis. In in-vitro setting, CCL26 triggers CX3CR1+ macrophages to secrete C1q in a CX3CR1 receptor-mediated manner. Furthermore, C1q and CCL26 are increased in BAL by allergen challenge in patients with allergic asthma. This data suggests CCL26 activates CX3CR1+ macrophages to facilitate efferocytosis via C1q secretion. In aim 2, we will examine the detailed mechanisms of CX3CR1+ macrophage activation through CCL26-mediated C1q secretion. Finally, we will extend the study to translational human research using the IRB-approved protocol for the segmental provocation with an allergen to evaluate the human relevance of the above proposed experiments. The proposed study is based on our strong supporting data on the new roles of CX3CR1+ macrophages in the resolution of allergic lung inflammation. This study will lead to a better understanding of the resolution process of allergic asthma.

标题：CX3CR1+巨噬细胞介导的嗜酸性过敏性肺注射的分辨率 抽象的： 最近的研究表明，组织居民巨噬细胞不仅参与炎症的倡议，而且参与 还可以解决局部炎症。精确确定巨噬细胞的子集 我们从事解决肺部感染并洞悉其功能，我们采用了新技术 质量细胞仪和单细胞RNA-seq（SC-RNA-Seq）分析人类和小鼠巨噬细胞 肺。我们的支持数据表明，肺泡巨噬细胞（AMS）在表型上是多种多样的，高度 响应过敏原挑战的动态。基于SC-RNA-seq数据，可以将AM聚集到几个 团体处于稳定状态。在这些组中，CX3CR1表达AM（CX3CR1+ AMS）的子集为 与经典居民AM相比 是CX3CR1负。在过敏性哮喘和哮喘小鼠模型的患者中，我们发现CX3CR1+ 过敏原挑战中BAL的AM明显增加。 CX3CR1+ AMS不仅表达 巨噬细胞和嗜酸性粒细胞标记物，例如人类Siglec-8。对CX3CR1-的进一步研究 记者和EPX-CRE（又称EO-CRE）记者小鼠揭示了CX3CR1+巨噬细胞吞噬了嗜酸菌 稳态和过敏性肺注射。在小鼠模型中CX3CR1+巨噬细胞的耗竭导致了 在自发性组织嗜酸性粒细胞稳定状态，并在过敏性肺中长时间组织嗜酸性粒细胞 炎。基于这些数据，我们假设新招募的CX3CR1+ AM子集促进了 清除组织嗜酸性粒细胞并促进过敏性肺注射的分辨率。在AIM 1中，我们将 专注于过敏性肺注射中CX3CR1+巨噬细胞的细胞动力学。关于 CX3CR1+介导的嗜酸性粒细胞清除的分子机制，我们检查了潜在的配体 CX3CR1 - CX3CL1和CCL26。我们发现CCL26在激活CX3CR1+中起关键作用 巨噬细胞，而CX3CL1是必不可少的。我们的SC-RNA-seq数据显示CX3CR1+ AM子集为 C1Q转录本的唯一来源 - 肿瘤的关键分子。在体外环境中，CCL26触发器 CX3CR1+巨噬细胞以CX3CR1受体介导的方式到秘密C1q。此外，C1Q和CCL26 过敏性哮喘患者的过敏原挑战在BAL中增加。该数据表明CCL26 激活CX3CR1+巨噬细胞，以通过C1Q分泌促进吞噬作用。在AIM 2中，我们将检查 通过CCL26介导的C1Q分泌，CX3CR1+巨噬细胞激活的详细机制。最后，我们 将扩展研究以使用IRB批准的方案进行分段来翻译人类研究 与过敏原一起挑衅以评估上述实验的人类相关性。 拟议的研究基于我们关于CX3CR1+巨噬细胞新作用的强大支持数据 过敏性肺注射的分辨率。这项研究将使人们对解决过程有更好的了解 过敏性哮喘。