Microbial Dysbiosis and Epithelial Dysfunction in Vitamin A-deficient Lungs

维生素 A 缺乏肺中的微生物失调和上皮功能障碍

• 批准号: 10607617
• 负责人: Kiloni Quiles
• 金额: $ 4.77万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10607617
• 关键词: Adult; Affect; Asthma; Biology; Bronchoalveolar Lavage Fluid; Cell Culture Techniques; Cells; Chronic Obstructive Pulmonary Disease; Collection; Communities; Complex; Core Facility; Data; Diet; Disease; Distal; Ecosystem; Environment; Epithelium; Event; Exhibits; Exposure to; Functional disorder; Future; Gene Expression; Genes; Health; Homeostasis; Human; Immune; Impairment; In Vitro; Injections; Knowledge; Laboratories; Link; Lower respiratory tract structure; Lung; Lung diseases; Lung infections; Maintenance; Malnutrition; Mammals; Measures; Mediating; Metabolic; Metabolic Pathway; Metabolism; Microbe; Microbial Biofilms; Microscope; Microscopy; Modeling; Mucociliary Clearance; Mucous Membrane; Mus; Natural regeneration; Outcome; Paraffin Embedding; Pathogenesis; Pathology; Pathway interactions; Patients; Pattern; Peer Review; Phenotype; Physiological Processes; Protocols documentation; Publishing; Pulmonary Pathology; Quantitative Reverse Transcriptase PCR; RNA; Respiratory Disease; Respiratory Tract Infections; Risk; Role; SARS-CoV-2 infection; Severities; Sterility; Structure of parenchyma of lung; System; Systems Biology; Techniques; Technology; Testing; Time; Tissue Stains; Tissues; Trachea; Tracheal Epithelium; Transcript; Tretinoin; Tuberculosis; Viral; Vitamin A; Vitamin A Deficiency; Work; airway epithelium; airway immune response; antimicrobial; cell motility; comparison control; design; dietary; differential expression; dysbiosis; epithelial injury; fluorescence imaging; germ free condition; gut dysbiosis; gut microbiome; host-microbe interactions; immune function; in silico; in vivo; interest; lung health; lung injury; lung microbiome; machine learning algorithm; metatranscriptomics; microbial; microbial colonization; microbial composition; microbiome; next generation sequencing; novel; permissiveness; programs; protein expression; protein structure; pulmonary function; repaired

Abstract Once believed to be sterile, recent studies now show microbes inhabiting healthy lungs that are dysregulated in patients with chronic obstructive pulmonary disease (COPD), asthma, tuberculosis (TB), and SARS-CoV-2 infection. Malnourished patients have an increased risk of respiratory infections and pathogenesis according to recent studies, indicating a potential link between metabolic status and lung homeostasis. These and other studies indicate the presence of host-microbe-vitamin A interactions that are present in the lung and influence the respiratory immune response, however details of whether changes to the lung microbiome is a cause or consequence of lung pathology, or whether retinoic acid (RA) – the bioactive metabolite of Vitamin A – can directly influence the lung microbiome in a host-independent manner are largely unknown. We hypothesize that dietary VAD-induced airway epithelial remodeling promotes microbial dysbiosis in the lung, further perpetuating respiratory epithelial dysfunction via host-microbe interactions. Aim 1 will determine whether dietary VAD directly alters metabolic pathways of opportunistic microbes found upregulated in the lower respiratory tract (LRT) of adult VAD mouse lungs. Our Microscopy core facilities offer state-of-the-art microscopes designed to capture high-quality fluorescent images that will enable us to identify microbial composition, protein expression, and localization within the airway (1.1). Using qRT-PCR machines freely accessible to me in the Pulmonary Center, we will measure changes in relative abundance of the opportunistic microbes upregulated in VAS and VAD lungs at 3 weeks and 8 weeks post-dietary modulation (1.2). Aim 2 will determine whether locally-derived vs. distally-derived metabolic changes in the host could influence airway epithelial remodeling in the lower respiratory tract. We will measure ciliary motility in tracheal explants of VAS and VAD lungs using a ciliary motility protocol already completed by our laboratory (2.1). We will also investigate whether microbes have the ability to influence epithelial remodeling and ciliary function independent of host metabolic status. We aim to expose VAS and VAD tracheal explants to microbes from opposing diets in-vitro (2.2) and repeat the ciliary motility technique proposed in Aim 1. Our lab’s metatranscriptomic work paired with the Pulmonary department’s proven expertise in intratracheal viral injections and cell culture techniques makes this sub-aim easily achievable within the proposed time frame. Aim 3 will identify mechanisms associated with microbial metabolic functions that are dysregulated in the absence of vitamin A. We will measure RNA transcripts of our upregulated microbes via qRT-PCR gathered from our explanted tissues mentioned in Aim 2 (3.1). We will also investigate microbe-microbe and microbe-environment interactions using an in-silico community-based modeling program called COMETS (3.2), a platform created by our collaborators at the BU Microbiome Initiative. The results of this work aims to uncover specific host-microbe and microbe-microbe interactions in VAD lungs that are essential for healthy lung function.

抽象的 一旦被认为是无菌的，最近的研究现在表明居住在健康肺部的微生物在 慢性阻塞性肺疾病（COPD），哮喘，结核病（TB）和SARS-COV-2患者 感染。根据呼吸道感染和发病机理的风险增加了。 最近的研究表明，代谢状况与肺稳态之间存在潜在的联系。这些和其他 研究表明存在肺中存在的宿主 - 微叶酸vitamin A相互作用和影响 呼吸免疫反应，但是有关肺微生物组的变化是原因还是 肺病理学的结果，或视黄酸（RA） - 维生素A的生物活性代谢产物 - 可以 直接以宿主独立的方式影响肺微生物组是未知的。我们假设这一点 饮食VAD诱导的气道上皮重塑促进肺中的微生物营养不良，进一步 通过宿主 - 微叶相互作用使呼吸上皮功能障碍永续存在。 AIM 1将确定是否 饮食VAD直接改变了在较低的机会中发现的机会性微生物的代谢途径 成年VAD小鼠肺的呼吸道（LRT）。我们的显微镜核心设施提供最先进的 显微镜旨在捕获高质量的荧光图像，这将使我们能够识别微生物 气道中的组成，蛋白质表达和定位（1.1）。自由使用QRT-PCR机器 我可以在肺中心接触，我们将衡量机会主义的相对丰度变化 在近期调节后3周零8周中，微生物在VAS和VAD肺中更新（1.2）。 AIM 2意志 确定本地衍生的与单独衍生的代谢变化是否可能影响气道 下呼吸道重塑。我们将测量VAS气管外植体中的睫状运动 使用睫状运动协议已经完成了我们的实验室（2.1），使用睫状运动方案。我们还将调查 微生物是否能够影响独立于宿主的上皮重塑和睫状功能 代谢状态。我们旨在将VAS和VAD气管外植体暴露于微生物中，以相反的饮食量 （2.2）并重复AIM 1提出的睫状运动技术。 肺部在气管内病毒注射和细胞培养技术方面的经过验证的专业知识使得 在拟议的时间范围内可以很容易地实现此子aim。 AIM 3将确定与 在没有维生素A的情况下失调的微生物代谢功能。我们将测量RNA转录本 我们通过QRT-PCR从AIM 2（3.1）中提到的外植物组织收集的QRT-PCR上调的微生物。我们 还将使用基于silico的社区的微生物微生物和微生物 - 环境相互作用 建模程序称为COMET（3.2），这是我们的合作者在BU Microbiome计划中创建的平台。 这项工作的结果旨在发现VAD肺中特定的宿主微生物和微生物相互作用 对于健康的肺功能至关重要。