Systems Biology of a MicroRNA Network in Neutrophilic Asthma

中性粒细胞性哮喘中 MicroRNA 网络的系统生物学

• 批准号: 10434046
• 负责人: Jose Luis Gomez-Villalobos
• 金额: $ 62.85万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10434046
• 关键词: Address; Adrenal Cortex Hormones; Affect; Airway Disease; Asthma; Biological; Biological Products; Cell Separation; Cells; Data Set; Environment; Epithelial Cells; Foundations; Future; Gene Expression; Gene Expression Regulation; Genes; Hospitalization; In Situ Hybridization; Inflammation; Inflammatory; Inhalation; Interleukin-17; Knowledge; Lead; Lymphocyte; Messenger RNA; MicroRNAs; Modality; New York; Pathway interactions; Patients; Public Health; Quality of life; Registries; Regulation; Role; Sampling; Severities; Severity of illness; Sputum; Steroid Resistance; Systems Biology; Therapeutic; Toll-Like Receptor Pathway; Transfection; Universities; Untranslated RNA; Work; airway epithelium; airway inflammation; airway obstruction; asthma exacerbation; cohort; experimental study; extracellular; extracellular vesicles; gene regulatory network; improved; improved outcome; member; miRNA expression profiling; neutrophil; novel; response; therapeutic development; therapeutic target; transcriptome sequencing; treatment response

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.

中性粒细胞气道炎症与哮喘严重程度增加有关。近50%的患者患有 哮喘是非嗜酸性的，因此不太可能对皮质类固醇治疗或电流产生完全反应 生物制品，说明了未得到满足的需求，以提高我们对这一群体患者的了解。尽管如此 对于中性粒细胞哮喘(NA)所涉及的通路，人们只有部分了解。MicroRNAs (MiRNAs)在基因调控中发挥强大的作用，并与T辅助分子2的气道炎症有关 得了哮喘。然而，在我们对miRNAs在NA中的作用的理解中存在着知识差距。 MiR-223-3p与严重的NA相关，并导致呼吸道上皮细胞变化，涉及NA中的miRNAs。 我们在耶鲁大学哮喘和呼吸道疾病队列中心(YCAAD)的研究小组发现miR-223-3p 属于痰细胞中十几个miRNA组成的网络，与气流阻塞、哮喘有关 住院时间、哮喘患者的生活质量、痰中淋巴细胞和中性粒细胞计数下降。至 为了了解这种细胞特异性的联系，我们进行了原位杂交来鉴定痰细胞表达 MiR-223-3p，发现中性粒细胞表达高水平的miR-223-3p。这种miRNA是正相关的 哮喘患者存在Toll样受体通路和IL-17的表达。 我们假设痰中的miRNA网络参与了Th17途径的调节，并 哮喘患者的中性粒细胞呼吸道炎症。我们将利用两个特征良好的队列，YCAAD和New 约克大学/贝尔维尤哮喘登记处(NYUBAR)，以实现以下目标： 目的1.确定痰中miRNA网络的纵向表达及其在中性粒细胞中的作用 呼吸道发炎。这一目标将决定miRNA在YCAAD队列中表达的稳定性，并将 验证miRNA网络在NYUBAR队列中的表达。 目的2.确定细胞外小泡中释放的网络miRNAs及其与中性粒细胞的关系 呼吸道发炎。这一目标将决定独特的网络miRNAs释放到 通过胞外小泡(EVS)的胞外环境，发挥其在Th17调节中的作用 呼吸道上皮细胞的炎症。 目的3.确定miRNA网络对呼吸道上皮细胞基因表达的影响及其作用 中性粒细胞的呼吸道炎症。这一目标将阐明网络中的miRNA如何与每个miRNAs相互作用 以及它们的调节作用如何汇聚在特定的炎症途径上。 这些研究将使用miRNA的新范例来研究一种知之甚少的哮喘内型。 中性粒细胞气道炎症的调节。该项目的成果将为下一步的 改进与NA相关的基因调控网络的识别，并导致潜在的治疗操作 中性粒细胞炎症中的miRNAs。