Extracellular vesicle-driven neutrophilic inflammation in cystic fibrosis lungs

囊性纤维化肺中细胞外囊泡驱动的中性粒细胞炎症

• 批准号: 10650427
• 负责人: Rabindra Marie-Jean Tirouvanziam
• 金额: $ 76.34万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10650427
• 关键词: Adult; Animals; Biogenesis; Bioinformatics; Biology; Cells; Chemotactic Factors; Child; Chromatin; Chronic; Citric Acid Cycle; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Cytoplasmic Granules; Data; Disease; Drug usage; Epithelium; European; Exocytosis; Functional disorder; Genetic Transcription; Genomics; Goals; HDAC11 gene; Histone Deacetylase Inhibitor; Human; Hyperactivity; Immunology; In Vitro; Inflammation; Intervention; Knowledge; Leukocytes; Licensing; Link; Liquid substance; Longevity; Lung; Medicine; Messenger RNA; Metabolic; Methods; Microbe; Mitochondria; Modeling; Mucous body substance; Mus; Mutation; Neutrophil Infiltration; Obstruction; Opportunistic Infections; Pathway interactions; Persons; Pharmaceutical Preparations; Pharmacotherapy; Play; Process; Production; Property; Proteins; Proteomics; Publishing; Pulmonary Cystic Fibrosis; Pulmonary Inflammation; RNA; Reaction; Reporting; Research; Role; Small Interfering RNA; Sputum; Surface; Testing; Therapeutic Intervention; Tissues; Transfection; Transfer RNA; Untranslated RNA; airway inflammation; autosome; c-myc Genes; cystic fibrosis airway; cystic fibrosis infection; cystic fibrosis patients; extracellular vesicles; human disease; human model; immunoregulation; in vivo; inhibitor; innovation; large scale production; migration; neutrophil; novel; pharmacologic; recruit; small molecule; stem; targeted treatment; tool; transcription factor; transcriptomics

PROJECT SUMMARY / ABSTRACT Neutrophils are the most abundant leukocyte subset in humans. Current therapies targeting neutrophils are limited to drugs that block their recruitment to tissues, leaving their functions mostly untouched. Our long-term goal is to develop new knowledge on neutrophil biology and function in humans and derive from it innovative treatments for conditions dominated by intractable neutrophil-driven inflammation, including cystic fibrosis (CF). CF is the most frequent autosomal disease in people of European descent, caused by mutations in the CFTR epithelial channel. Recent research has shown that CFTR dysfunction causes local mucus stasis obstruction, followed by neutrophil recruitment, and paradoxically, opportunistic infections. This paradox stems from the fact that most neutrophils recruited to CF airways in vivo undergo hyperactive granule release, and become immunomodulatory and metabolically licensed, yet fail to kill microbes, in a functional fate we dubbed “GRIM”. Using an organotypic model of human airway inflammation, we showed previously that naïve blood neutrophils can be made to acquire the GRIM fate upon transepithelial migration in vitro into airway fluid from CF patients. Further preliminary studies of GRIM adaptation presented here support an obligate upstream role for extracellular vesicles (EVs) present in airway fluid carrying the long non-coding RNA MALAT-1, and the downstream implication of transcriptional firing and mitochondrial rewrting, with histone deacetylase 11 (HDAC11) and cMyc as key modulators. Remarkably, we show that GRIM neutrophils produced in vitro secrete secondary EVs that can in turn induce GRIM adaptation in a new wave of neutrophils. Also, we show that siRNA against MALAT-1 and HDAC11 and inhibition of HDAC11 by the small molecule drug SIS17 normalize granule release and bacterial killing by GRIM neutrophils, as well as their release of secondary EVs. Our objective is to fully characterize mechanisms of GRIM adaptation in CF airway neutrophils. Our central hypothesis is that chronic CF airway inflammation is a feed-forward process in which resident GRIM neutrophils release EVs that induce the GRIM fate in newly recruited neutrophils. This study introduces a new and innovative basic paradigm of neutrophil-driven human lung inflammation with key roles for EVs, MALAT-1, HDAC11 and cMyc, and innovative methods to deconstruct this paradigm (model for mass production of lung-recruited neutrophils, optimized tools to characterize lung EVs, and drug- / RNA / siRNA interventions targeted at neutrophils). We expect this study to yield transformative findings in our basic understanding and ability to treat chronic neutrophilic inflammation and infection in CF and other diseases.

项目总结/摘要 中性粒细胞是人类中最丰富的白细胞亚群。目前靶向中性粒细胞的疗法是 仅限于阻止它们被组织吸收的药物，使它们的功能基本上不受影响。我们的长期 目标是发展人类中性粒细胞生物学和功能的新知识，并从中获得创新 治疗以难治性嗜酸性粒细胞驱动的炎症为主的疾病，包括囊性纤维化（CF）。 CF是欧洲血统人群中最常见的常染色体疾病，由CFTR突变引起 上皮通道最近的研究表明，CFTR功能障碍导致局部粘液淤滞阻塞， 其次是中性粒细胞聚集，矛盾的是，还有机会性感染。这个悖论源于 事实上，大多数中性粒细胞募集到CF气道在体内经历过度活跃的颗粒释放， 免疫调节和代谢许可，但不能杀死微生物，在一个功能的命运，我们称之为“GRIM”。 使用人体气道炎症的器官型模型，我们先前表明，幼稚的血液中性粒细胞， 可以使其在体外经上皮迁移到CF患者的气道液中时获得GRIM命运。 进一步的初步研究GRIM适应这里提出支持一个强制性的上游作用， 存在于气道液中的携带长非编码RNA MALAT-1的细胞外囊泡（EV），以及 组蛋白去乙酰化酶11对转录激活和线粒体逆转的下游影响 （HDAC 11）和cMyc作为关键调节剂。值得注意的是，我们发现GRIM中性粒细胞在体外产生， 分泌继发性EV，继而在新一波中性粒细胞中诱导GRIM适应。此外，我们显示 针对MALAT-1和HDAC 11的siRNA以及小分子药物SIS 17对HDAC 11的抑制 使GRIM中性粒细胞的颗粒释放和细菌杀伤以及它们的继发性EV释放正常化。 我们的目标是充分表征CF气道中性粒细胞中GRIM适应的机制。我们的中央 假设慢性CF气道炎症是一个前馈过程，其中居民GRIM 中性粒细胞释放EV，其在新募集的中性粒细胞中诱导GRIM命运。 这项研究介绍了一个新的和创新的基本范式，嗜中性粒细胞驱动的人类肺部炎症， EV、MALAT-1、HDAC 11和cMyc的关键作用，以及解构这种范式的创新方法（模型 用于大量生产肺募集的中性粒细胞，表征肺EV的优化工具，以及药物- / RNA / 靶向中性粒细胞的siRNA干预）。我们希望这项研究能在我们的基础研究中产生变革性的发现。 了解并有能力治疗慢性嗜酸性炎症和CF感染及其他疾病。