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.

呼吸道粘液清除是一种主要的肺防御机制，在新生儿早期 代表着肺部健康的一个关键的“易感窗口”。围产期脑白质疏松主要成分的变化 粘液清除系统已被定性地描述，但这些系统的细节和功能意义 变化是未知的。我们在小鼠和人类身上的初步数据表明：1)呼吸道粘液成分是 发育调节；和2)粘液清除系统的成分受 早期生命中的宿主微生物群。我们已经确定了一种可测试的机制，将宿主微生物区系与 新生儿呼吸道粘液生物学与肺内稳态，即IL-22+ILC3介导的肠肺轴。我们也 假设人类在促进呼吸道粘液清除的关键因素上表现出年龄相关的变化， 例如，MUC5AC级别。为了验证这些假设，我们提出了以下目标：1)测试宿主 微生物区系失调或阻断IL-22+ILC3介导的肠肺轴会影响丰度， 新生小鼠与成年小鼠呼吸道粘液的组成和功能。无菌饲养的同基因小鼠 或者无特定病原体的情况，无论是幼稚的还是用抗生素治疗的，都将在出生后一天进行评估。 (PND)10和PND66用于：1)支气管肺泡灌洗(BAL)蛋白成分、粘液浓度和 分泌性粘蛋白定量；2)呼吸道粘液纤毛清除；3)肺mRNA测序。特定的 将评估IL-22信号和结果在呼吸道表面脱水方面的作用 分别用IL-22KO小鼠和Scnn1b-TG小鼠。宿主微生物区系的变化将由以下因素决定 细菌16S核糖体RNA基因定量。2)检测类CF型宿主微生物区系失调 影响正常的IL-22+ILC3介导的新生小鼠肠-肺轴的建立。同源基因 ∆F508CF小鼠和WT仔鼠，无论是幼稚的还是用抗生素治疗的，都将在PND10上用于研究 IL22+ILC3细胞流入肺的情况参考肠道微生物区系的定量分析。与之平行的是肺 并收集BAL样本，以评估CF肠道生物失调对早期肺转录组和 新生儿呼吸道分泌物的组成。与Scnn1b-TG小鼠的杂交将被用来加剧 ∆F508CF型肺表型。3)测试“健康”呼吸道粘液的组成是否呈现年龄- 人类之间依赖的、数量上的差异。从气管插管采集的呼吸道粘液样本 用于新生儿/婴儿/儿童(0-5岁)和成人(25-40岁)，无以下病史或症状 将对呼吸道疾病进行粘蛋白和蛋白质成分分析。组织学标本取自 《龙图》 生物信息库(BRINDL) 将探索特定粘液的mRNA和蛋白质表达 记号笔。完成这些目标将建立一个新的概念框架，并提供亟需的 研究发病机制、开发治疗方法和解释宿主的实验证据 微生物区系对“早期”肺部健康和早发性粘液阻塞性肺疾病的影响。