MicroRNA Networks in self-sustaining type 2 airway niches in asthma

项目摘要 Th 2高哮喘被定义为持续的气道炎症，其由细胞因子之间的串扰驱动。 上皮细胞和组织驻留的先天性和适应性免疫细胞。我们合作的中心主题 一项研究计划认为，哮喘中发生的2型炎症的局部性质反映了哮喘的发展， 持续性2型气道小生境含有重编程上皮细胞和免疫细胞。破坏这些生态位 可能持久影响哮喘发病机制。在正常情况下，瞬态2型响应的操作是 维持上皮屏障功能。在哮喘中，抑制这些反应的调节机制失效， 伴有上皮细胞重编程和粘蛋白高分泌的2型炎症持续存在于 航空公司.该项目的中心目标是确定细胞重编程的分子决定因素， Th 2-高哮喘持续免疫病理学。 拟议的研究集中在两种细胞类型，其编程直接影响这一病理过程： 调节性T（Treg）细胞和气道上皮细胞。我们的方法建立在初步数据的基础上， 2型炎症和肺功能不全与气道THBG亚群的频率呈负相关， 对控制TcB和上皮细胞编程的不同miRNA家族的阳性鉴定。 该项目分为三个目标：在目标1中，我们将使用单细胞测序，mRNA和miRNA分析 和质量细胞计数来定义气道Treg群体，并比较持续性气道2型中的Treg亚群， 以粘液嵌塞的病灶部位为标志的小生境与那些填充未受影响的气道的小生境。在目标2中，我们 使用miRNA指导的途径发现来剖析Treg编程，这是一种新的实验框架， miRNA：靶基因网络。类似的方法将在Aim 3中应用于人气道上皮细胞， 揭示miRNA：控制细胞重编程和标记气道的粘蛋白高分泌的靶向网络 2型哮喘的小生境。如果成功的话，这项研究将发现新的基因和途径， 哮喘病理学，推进了我们对免疫调节机制如何在2型气道中失效的理解 壁龛，并提出策略，以破坏炎症，维持这种疾病。

Project Summary Th2 high asthma is defined by persistent airway inflammation that is driven by cytokine crosstalk between epithelial cells and tissue resident innate and adaptive immune cells. The central theme of our collaborative research program is that the focal nature of type 2 inflammation that occurs in asthma reflects the development of persistent type 2 airway niches containing reprogrammed epithelial and immune cells. Disrupting these niches may durably impact asthma pathogenesis. Under normal conditions, transient type 2 responses operate to maintain epithelial barrier function. In asthma, regulatory mechanisms that dampen these responses fail, and type 2 inflammation with epithelial cell reprogramming and mucin hypersecretion persists at focal sites in the airways. The central objective of this project is to define molecular determinants of cell reprogramming that sustain persistent immunopathology in Th2-high asthma. The proposed studies focus on two cell types whose programming directly affects this pathological process: T regulatory (Treg) cells and airway epithelial cells. Our approach builds upon preliminary data indicating that type 2 inflammation and lung dysfunction correlate inversely with the frequency of a subset of airway Tregs, and the positive identification of distinct miRNA families that control the programming of Tregs and epithelial cells. The project is organized into three aims: In Aim 1, we will use single cell sequencing, mRNA and miRNA profiling and mass cytometry to define airway Treg populations, and to compare Treg subsets in persistent airway type 2 niches marked by focal sites of mucus impaction with those that populate unaffected airways. In Aim 2, we will dissect Treg programming using miRNA-directed pathway discovery, a novel experimental framework for probing miRNA:target gene networks. A similar approach will be applied in human airway epithelial cells in Aim 3 to reveal miRNA:target networks that control the cell reprogramming and mucin hypersecretion that marks airway type 2 niches in asthma. If successful, the proposed research will uncover novel genes and pathways critical to the pathology of asthma, advance our understanding how immune regulation mechanisms fail in type 2 airway niches, and suggest strategies to disrupt the inflammation that sustain this disease.