Host microbiota and airway mucus: a novel paradigm for lung defense and immune homeostasis during early post-natal development

宿主微生物群和气道粘液：产后早期发育期间肺防御和免疫稳态的新范例

• 批准号: 10535465
• 负责人: Alessandra Maria Livraghi-Butrico
• 金额: $ 38.88万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10535465
• 关键词: 5 year old; Ablation; Adult; Affect; Age; Animals; Antibiotic Therapy; Antibiotics; Asthma; Bacterial Infections; Bacterial Pneumonia; Biology; Bronchoalveolar Lavage; Cell Differentiation process; Cells; Child; Congenic Mice; Cystic Fibrosis; Data; Defense Mechanisms; Dehydration; Development; Disease; Elements; Epithelial Cells; Exhibits; Gene Expression; Germ-Free; Harvest; Homeostasis; Human; Immune; Infant; Inflammatory; Inhalation; Knockout Mice; Knowledge; Life; Link; Lung; Lung immune response; MUC5AC gene; Mediating; Messenger RNA; Modeling; Mucins; Mucociliary Clearance; Mucous body substance; Mus; Neonatal; Operative Surgical Procedures; Organ; Outcome; Pathogenesis; Pattern; Perinatal; Phenotype; Predisposition; Proteins; Publications; Pulmonary Cystic Fibrosis; RNA, Ribosomal, 16S; Recording of previous events; Respiratory Disease; Respiratory Signs and Symptoms; Respiratory System; Sampling; Secretory Cell; Shapes; Signal Transduction; Surface; Syndrome; System; Testing; Therapeutic; Tube; Wild Type Mouse; age related; biobank; congenic; cystic fibrosis mouse; dysbiosis; early onset; endotracheal; germ free condition; gut dysbiosis; gut microbiota; gut-lung axis; histological specimens; host microbiome; host microbiota; interleukin-22; lung health; lung microbiota; mRNA Expression; mRNA sequencing; mucus clearance; mucus-associated lung diseases; neonatal mice; neonatal period; neonate; novel; pediatric patients; postnatal; postnatal development; postnatal period; protein expression; rRNA Genes; transcriptome

Airway mucus clearance is a primary lung defense mechanism and the early neonatal period represents a critical “window of susceptibility” for lung health. Perinatal changes in key components of the mucus clearance system have been qualitatively described, but the details and functional significance of these changes are unknown. Our preliminary data in mice and humans indicate that: 1) airway mucus composition is developmentally regulated; and 2) components of the mucus clearance system are affected by the status of the host microbiota in early life. We have identified a testable mechanism to link the host microbiota with neonatal airway mucus biology and lung homeostasis, i.e., the IL-22+ILC3-mediated gut-lung axis. We also posit that humans exhibit age-dependent changes in key elements contributing to airway mucus clearance, e.g., MUC5AC levels. To test these hypotheses, we propose the following aims: 1) To test whether host microbiota dysbiosis or blockade of the IL-22+ILC3-mediated gut-lung axis affect the abundance, composition, and function of airway mucus in neonatal vs. adult mice. Congenic mice raised in germ-free or specific-pathogen free conditions, either naïve or treated with antibiotics, will be assessed at post natal day (PND)10 and PND66 for: 1) bronchoalveolar lavage (BAL) protein composition, mucus concentration, and secreted mucins quantification; 2) airway mucociliary clearance; and 3) lung mRNA sequencing. Specific contributions of IL-22 signaling and outcomes in the context of airway surface dehydration will be assessed using IL-22 KO mice and Scnn1b-Tg mice, respectively. Changes in host microbiota will be determined by bacterial 16S ribosomal RNA gene quantification. 2) To test whether CF-like host microbiota dysbiosis affects the establishment of the normal, IL-22+ILC3-mediated gut-lung axis in neonatal mice. Congenic ∆F508 CF mice and WT littermates, either naïve or treated with antibiotics, will be used at PND10 to study the influx of IL22+ILC3 cells into the lung referenced to quantitative analysis of their gut microbiota. In parallel, lung and BAL samples will be collected to assess the effect of CF gut dysbiosis on the early lung transcriptome and the composition of neonatal airway secretions. Crosses with Scnn1b-Tg mice will be used to exacerbate the ∆F508 CF lung phenotype. 3) To test whether the composition of “healthy” airway mucus presents age- dependent, quantitative differences in humans. Airway mucus samples harvested from endotracheal tubes used in neonates/infants/children (0-5 years old) and adults (25-40 years old) with no history or symptoms of respiratory diseases will be analyzed for mucin and protein composition. Histologic specimens obtained from the LungMAP Biorepository (BRINDL) will be probed for mRNA and protein expression of specific mucus markers. Completion of these aims will establish a new conceptual framework and provide much needed experimental evidence to study the pathogenesis, develop therapeutic approaches, and account for host microbiota influences on “early” lung health and early onset muco-obstructive lung diseases.

气道粘液清除是主要的肺防御机制，是新生儿早期 代表着肺部健康的关键“易感性”。关键组成部分的围产期变化 粘液清除系统已被定性地描述，但是这些细节和功能意义 变化未知。我们在小鼠和人类中的初步数据表明：1）气道粘液组成是 开发监管； 2）粘液清除系统的组件受状态的影响 早期的寄主微生物群。我们已经确定了一种可测试机制，可以将宿主微生物群与 新生儿气道粘液生物学和肺稳态，即IL-22+ILC3介导的肠肺轴。我们也是 假定人类在关键因素中表现出年龄的变化，导致气道粘液清除率， 例如，MUC5AC水平。要检验这些假设，我们提出以下目的：1）测试主机是否 IL-22+ILC3介导的肠道轴的微生物群或封锁会影响抽象， 新生儿与成年小鼠中气道粘液的组成和功能。无菌饲养的先天小鼠 或将在出生日期进行评估，或用抗生素治疗的特异性无病原疾病 （PND）10和PND66：1）支气管肺泡灌洗（BAL）蛋白质成分，粘液浓度和 分泌的粘蛋白定量； 2）气道粘膜扫膜清除； 3）肺mRNA测序。具体的 将评估IL-22信号传导和结果的贡献 分别使用IL-22 KO小鼠和SCNN1B-TG小鼠。宿主微生物群的变化将由 细菌16S核糖体RNA基因定量。 2）测试是否像CF一样宿主微生物群营养不良 影响正常的IL-22+ILC3介导的新生儿小鼠中的肠道轴。高等人物 ∆F508 CF小鼠和WT同窝仔，幼稚或用抗生素治疗，将在PND10使用 将IL22+ILC3细胞涌入肺中，参考了其肠道菌群的定量分析。并联，肺 并将收集BAL样品，以评估CF肠道营养不良对早期肺转录组和 新生儿气道分泌物的组成。用SCNN1B-TG小鼠的十字架将用于加剧 ∆F508 CF肺表型。 3）测试“健康”气道粘液的组成是否呈现年龄 - 人类的依赖性，定量差异。从气管管中收获的气道粘液样品 用于新生儿/婴儿/儿童（0-5岁）和成人（25-40岁），没有病史或症状 呼吸道疾病将分析粘蛋白和蛋白质组成。从 肺部 生物座席（Brindl） 将探测特异性粘液的mRNA和蛋白质表达 标记。这些目标的完成将建立一个新的概念框架，并提供急需的 研究发病机理，开发治疗方法并说明宿主的实验证据 微生物群会影响“早期”肺部健康和早期发作性粘液刺激性肺部疾病。