Systems Biology of a MicroRNA Network in Neutrophilic Asthma

嗜中性粒细胞气道炎症与哮喘严重程度增加相关。近50%的患者 哮喘是非嗜酸性粒细胞性的，因此不太可能对皮质类固醇治疗或目前的 生物制剂，说明了未满足的需要，以提高我们对这组患者的了解。尽管有这些 尽管哮喘与哮喘的发病相关，但涉及嗜酸性哮喘（NA）的途径仅部分了解。微rna miRNAs在基因调控中发挥着强大的作用，并与辅助性T细胞2气道炎症有关。 哮喘病然而，我们对miRNA在NA中的作用的理解存在知识差距。 miR-223- 3 p与严重NA相关，并导致气道上皮改变，涉及NA中的miRNA。 我们在耶鲁大学哮喘和气道疾病队列中心（YCAAD）的研究小组发现，miR-223- 3 p 属于痰细胞中十几种miRNAs的网络，与气流阻塞、哮喘 住院、哮喘生活质量下降、痰中淋巴细胞和中性粒细胞计数。到 为了了解这种细胞特异性的联系，我们进行了原位杂交，以确定痰细胞表达 miR-223- 3 p，并发现中性粒细胞表达高水平的miR-223- 3 p。这种小RNA正相关 哮喘患者Toll样受体通路和IL-17的表达。 我们假设痰液中的miRNA网络参与了Th 17通路的调节， 哮喘中的嗜酸性气道炎症。我们将利用两个特征良好的队列，YCAAD和新的 约克大学/贝尔维尤哮喘登记处（NYUBAR），以实现以下目标： 目标1。为了确定痰液miRNA网络的纵向表达及其在嗜酸性粒细胞中的作用， 气道炎症这一目标将确定YCAAD队列中miRNA表达的稳定性， 验证NYUBAR队列中miRNA网络的表达。 目标2.为了确定细胞外囊泡中释放的网络miRNAs及其与嗜酸性粒细胞的关系， 气道炎症这一目标将决定独特的网络miRNAs被释放到细胞中的程度。 通过细胞外囊泡（EV）调节细胞外环境，并发挥其在Th 17调节中的功能作用 气道上皮细胞炎症。 目标3。确定miRNA网络在气道上皮基因表达中的作用及其贡献 嗜肺性气道炎症这一目标将阐明网络中的miRNAs如何与每一个相互作用。 以及它们的调节作用如何汇聚在特定的炎症通路上。 这些研究将使用一种新的miRNA范式来研究一种知之甚少的哮喘内型 调节嗜酸性气道炎症。该项目的成果将为 改进了NA相关基因调控网络的鉴定，并导致潜在的治疗操作 在嗜肺性气道炎症中的作用。

Neutrophilic airway inflammation is associated with increased asthma severity. Nearly 50% of patients with asthma are non-eosinophilic and thus less likely to respond completely to corticosteroid therapy or current biologics, illustrating the unmet need to improve our understanding of this group of patients. Despite these associations, the pathways involved in neutrophilic asthma (NA) are only partially understood. MicroRNAs (miRNAs) exert a powerful effect on gene regulation and have been implicated in T helper 2 airway inflammation in asthma. However, there is a knowledge gap in our understanding of the role of miRNAs in NA. MiR-223-3p is associated with severe NA and leads to airway epithelial changes, implicating miRNAs in NA. Work by our group at the Yale Center for Asthma and Airway Disease cohort (YCAAD) found that miR-223-3p belongs to a network of a dozen miRNAs in sputum cells associated with airflow obstruction, asthma hospitalizations, decreased asthma quality of life, lymphocyte and neutrophil counts in the sputum. To understand this cell-specific association we performed in situ hybridization to identify sputum cells expressing miR-223-3p and found that neutrophils express high levels of miR-223-3p. This miRNA is positively correlated with toll-like receptor pathways and IL-17 expression in patients with asthma. We hypothesize that the sputum miRNA network is involved in the regulation of the Th17 pathway and neutrophilic airway inflammation in asthma. We will utilize two well characterized cohorts, YCAAD and the New York University/Bellevue Asthma Registry (NYUBAR), to execute the following aims: Aim 1. To determine the longitudinal expression of the sputum miRNA network and its role in neutrophilic airway inflammation. This aim will determine the stability of miRNA expression in the YCAAD cohort and will validate the expression of the miRNA network in the NYUBAR cohort. Aim 2. To define the network miRNAs released in extracellular vesicles and their association with neutrophilic airway inflammation. This aim will determine the extent to which unique network miRNAs are released into the extracellular environment via extracellular vesicles (EVs) and exert their functional role in the regulation of Th17 inflammation in airway epithelial cells. Aim 3. To determine the effect of the miRNA network in airway epithelial gene expression and its contribution to neutrophilic airway inflammation. This aim will elucidate how the miRNAs in the network interact with each other and how their regulatory role converges on specific inflammatory pathways. These studies will investigate a poorly understood asthma endotype using a novel paradigm of miRNA regulation of neutrophilic airway inflammation. The results derived from this project will lay the foundation for the improved identification of gene regulatory networks involved in NA and lead to potential therapeutic manipulation of miRNAs in neutrophilic airway inflammation.