Transcriptional Regulation of Migrating Neutrophils during Pneumonia

肺炎期间迁移中性粒细胞的转录调节

• 批准号: 10642707
• 负责人: Katrina Traber
• 金额: $ 41.25万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10642707
• 关键词: Address; Affect; Air; Alveolar; Automobile Driving; Biology; Bone Marrow; COVID-19 pandemic; Cell Separation; Cell membrane; Cells; Circulation; Clinical; Cytokine Signaling; Data; Development; Environment; Epithelium; Equilibrium; Exposure to; Flow Cytometry; Future; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Heterogeneity; Immune System Diseases; Immune response; Immunologics; In Vitro; Infection; Inflammatory; Injury; Intervention; Knowledge; Label; Lead; Liquid substance; Location; Lung; Maintenance; Mediating; Metabolism; Modeling; Morbidity - disease rate; Mucous Membrane; Mus; Outcome; Pathway interactions; Pneumonia; Population; Proteins; Recording of previous events; Site; Sorting; Stimulus; Surface; Testing; Therapeutic; Thinness; Tissues; Transcriptional Regulation; Travel; Up-Regulation; Work; biomarker identification; in vivo; in vivo Model; loss of function; migration; mortality; neutrophil; novel; novel marker; pathogen; pneumonia model; programs; response; single-cell RNA sequencing; transcription factor; transcriptome; transcriptome sequencing; transcriptomics

Project Summary/Abstract Pneumonia remains a significant cause of morbidity and mortality, which has come into sharp focus recently during the COVID-19 pandemic. The lung is immunologically unique, and neutrophils traversing the lungs must be ready to respond to potential pathogen challenges without causing injury to surrounding tissues. Disruptions of this balance lead to many severe complications of pneumonia, but we lack therapeutics directed toward neutrophil-mediated immune dysfunction. Our preliminary data demonstrate that neutrophils undergo transcriptional remodeling as they travel from the circulation to the alveolar space during pneumonia. A subset of pro-inflammatory genes is upregulated as neutrophils move from the vasculature to the interstitium, while genes related to metabolism and degranulation are upregulated as cells move from interstitium to the alveolar space. Maintenance of appropriate neutrophil function and development requires tight regulation of gene changes, yet the specific pathways underlying such changes constitute a major knowledge gap. The goal of this proposal is to delineate the transcriptional and functional changes neutrophils undergo as they migrate from the circulation to the site of infection during pneumonia. Our central hypothesis is that during pneumonia, appropriate neutrophil function is achieved by proscribed responses to specific environmental stimuli, resulting in a heterogenous population defined by their interaction history. Aim 1: Test the hypothesis that location-specific neutrophil responses in pneumonia are achieved by the tuning of neutrophil transcriptome and function by environmental stimuli. Using complementary in vitro and in vivo models we will determine the contributions of migration, pathogen interaction and cytokine signaling to neutrophil gene programs and function. Outcomes will include transcriptional and functional changes predicted by our preliminary studies. These findings will then be confirmed in an in vivo model of pneumonia, employing exogenously added labeled neutrophils. Aim 2: Test the hypothesis that during transit to the alveolar space, neutrophils differentiate into subpopulations unique to the lung, with transcriptional and functional heterogeneity. Using a combination of in vivo cell labeling, flow cytometry assisted cell sorting, and single cell RNA sequencing, we will track modulations in the neutrophil transcriptome from circulation to alveolar space at a single cell level. Aim 3: NFκB-RelA is a central transcription factor in neutrophils, driving the upregulation of a pro-inflammatory transcriptional program during migration to the alveolar space. Using complementary in vitro and in vivo loss-of function models we will determine the effect of neutrophil NFκB-RelA on pneumonia outcomes neutrophil transcriptome and neutrophil function. The results of these studies will have a positive impact the field of pneumonia biology furthering our knowledge of neutrophil function and laying the groundwork for future development of novel host-directed neutrophil-based pneumonia therapeutics.

项目概要/摘要 肺炎仍然是发病和死亡的一个重要原因，最近已成为人们关注的焦点 在 COVID-19 大流行期间，肺部具有独特的免疫功能，中性粒细胞必须穿过肺部。 准备好应对潜在的病原体挑战，而不会对周围组织造成伤害。 这种平衡的破坏会导致肺炎的许多严重并发症，但我们缺乏针对这些并发症的治疗方法 我们的初步数据表明，中性粒细胞介导的免疫功能障碍。 肺炎期间从循环系统移动到肺泡腔时的转录重塑。 当中性粒细胞从脉管系统移动到间质时，促炎基因的表达上调，而 当细胞从间质移动到肺泡时，与代谢和脱颗粒相关的基因上调 维持适当的中性粒细胞功能和发育需要基因的严格调控。 变化，但这些变化背后的具体途径构成了主要的知识差距。 该提案旨在描述中性粒细胞在迁移时经历的转录和功能变化 我们的中心假设是在肺炎期间从循环到感染部位。 肺炎，适当的中性粒细胞功能是通过对特定的禁止反应来实现的 环境刺激，导致由其相互作用历史定义的异质群体。 目标 1：检验肺炎中位置特异性中性粒细胞反应是通过调节实现的假设 通过环境刺激研究中性粒细胞转录组和功能。 我们将通过模型确定迁移、病原体相互作用和细胞因子信号传导的贡献 结果将包括预测的中性粒细胞基因程序和功能变化。 通过我们的初步研究，这些发现将在肺炎体内模型中得到证实。 外源添加标记的中性粒细胞 目标 2：检验在转运至肺泡腔期间， 中性粒细胞分化为肺特有的亚群，具有转录和功能 结合使用体内细胞标记、流式细胞术辅助细胞分选和单细胞。 RNA 测序，我们将跟踪中性粒细胞转录组从循环到肺泡空间的调节 目标 3：NFκB-RelA 是中性粒细胞中的核心转录因子，驱动 a 的上调。 使用互补的体外迁移过程中的促炎转录程序。 和体内功能丧失模型，我们将确定中性粒细胞 NFκB-RelA 对肺炎的影响 结果 中性粒细胞转录组和中性粒细胞功能将产生积极的结果。 影响肺炎生物学领域，加深我们对中性粒细胞功能的了解并奠定基础 为未来开发新型基于宿主的中性粒细胞肺炎疗法奠定了基础。