Testing the hypothesis that microbial energetic hijacking of the CF immune response selects for specific pathogens during lung function decline

检验以下假设：CF 免疫反应的微生物能量劫持会在肺功能下降期间选择特定病原体

• 批准号: 10175023
• 负责人: Dianne K Newman
• 金额: $ 53.11万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10175023
• 关键词: Adult; Anaerobic Bacteria; Animal Model; Antibiotics; Automobile Driving; Bacteria; Bacterial Physiology; Carbon; Cell Respiration; Charge; Chronic; Complex; Consumption; Cystic Fibrosis; Cystic Fibrosis sputum; Disease Progression; Dissection; Drug Design; Drug Metabolic Detoxication; Earth science; Energy-Generating Resources; Environment; Enzymes; Epithelial Cells; Fingerprint; Floods; Forensic Medicine; Gases; Gene Expression; Generations; Genes; Genetic; Growth; Habitats; Health; Human; Hypoxia; Image; Immune response; Immune system; In Vitro; Individual; Infection; Inflammatory; Isotopes; Knowledge; Learning; Lung infections; Mass Spectrum Analysis; Measurable; Measurement; Measures; Membrane; Metabolic; Metabolism; Microbe; Microbial Biofilms; Molecular Profiling; Morbidity - disease rate; Mucous body substance; Nitric Oxide; Nitrous Oxide; Oxidants; Oxides; Oxygen; Pathway interactions; Patients; Pattern; Physiological; Pilot Projects; Play; Process; Production; Respiratory Failure; Respiratory Process; Role; Site; Soil; Source; Sputum; System; Testing; Tobramycin; antibiotic tolerance; cohort; combat; cystic fibrosis airway; cystic fibrosis infection; cystic fibrosis patients; denitrification; dysbiosis; experimental study; fitness; fungus; host-microbe interactions; in situ imaging; in vivo; innovation; insight; member; microbial; microbial community; microbiome; microbiota; mortality; neutrophil; novel; opportunistic pathogen; oxidation; pathogen; pulmonary function decline; stable isotope; tool; trait; translational approach

PROJECT SUMMARY Individuals living with cystic fibrosis (CF) combat devastating, chronic microbial infections of the airways. Though CF patients are usually colonized by otherwise innocent bacteria and fungi—many of which derive from soil environments—the compromised CF immune system is unable to clear these opportunists, and the ensuing struggle between host and microbes eventually leads to failure of the pulmonary system. Conventional antibiotics are not very effective. One widely held view for why this is the case is that the resident microbiota are growing slowly and their membranes are insufficiently charged to take up commonly-used antibiotics such as tobramycin. Accordingly, if we seek new ways to treat CF lung infections, we must better understand how microbes thrive in this habitat. Different lines of evidence indicate that oxygen is limiting in the CF airways at the microscale relevant to opportunistic pathogens. Though carbon sources are replete in the mucus-filled airways, in the absence of oxygen, the bacteria and fungi that come to dominant this habitat must employ alternative energy generation strategies to aerobic respiration. How do they do this? Energy flow is the driving organizer of any microbial community, including those found in the CF airways. It is well established that the nitric oxide (NO) generated by the immune system in the CF patients is lower than that made by healthy individuals. Indeed, epithelial cells are severely compromised in NO production, yet neutrophils that flood the CF airways can still generate NO; this NO is insufficient to kill the microbes, but has the potential to transform into an energy source for those capable of denitrification. Accordingly, we seek to test the hypothesis that microbial energetic hijacking of the CF immune response via denitrification pathways selects for specific pathogens during lung function decline. To test this hypothesis, we plan to leverage powerful isotopic tools from the Earth sciences that permit the sources of metabolites in complex environments, such as the CF airways, to be determined atom by atom, providing a non-invasive, forensic molecular fingerprint. In particular, we will focus on interpreting N2O, a measurable product of the denitrification pathway in CF sputum and breath gas. In addition to approaches employing bacterial physiology, genetics, and in situ imaging of host-microbe interactions, we will use these isotopic tools to gain insight into how the NO that is reduced to N2O in the CF airways may favor the fitness of particular microbial community members. To do this, we propose three specific aims: Aim 1 will identify the conditions and pathways leading to N2O production by common CF bacteria and fungi, Aim 2 will utilize advanced isotopic analyses to dissect the N2O produced by these microbes to forensically infer its source, and Aim 3 will apply these insights to a pilot study of adult CF patients to determine whether particular types of denitrifiers are favored as lung function declines. Attainment of these objectives will provide the critical knowledge needed to guide early translational approaches for treating infections of the CF airways, as well as establish tools that can be applied more broadly to other studies of dysbiosis.

项目摘要 患有Cystem纤维化（CF）战斗毁灭性的人，气道的慢性微生物感染。 尽管CF患者通常被其他无辜的Bactteria和Fungi殖民 - 其中许多 从土壤环境中 - 受损的CF免疫系统无法清除这些机会主义者，并且 随之而来的宿主和微生物之间的斗争最终导致肺系统失败。 抗生素不是很有效。 正在缓慢增长，其膜不足以承担通常使用的抗体药 相应地，如果我们看到治疗CF肺部感染的方法 微生物在这个栖息地繁衍生息。 与机会性病原体相关的显微镜。 气道在没有氧气的情况下，占主导地位的栖息地的bactteria和真菌必须采用 有氧呼吸的替代能源产生策略。 任何微生物社区的组织者，包括在CF Airways中发现的组织者。 CF患者中免疫系统产生的一氧化氮（NO）较低 个人。 CF航空仍然可以生成不 成为能够剥离的人的能量来源。 通过非硝化途径对CF免疫反应的微生物能量劫持选择特定 肺功能下降期间的病原体。 从允许在复杂环境中代谢物的科学（例如CF） 通过原子确定原子，尤其是非侵入性的法医指纹。 我们将专注于解释N2O，N2O是CF痰液和呼吸中硝化途径的可测量产物 除了采用细菌生理学，遗传学和原位成像的方法之外 互动，我们将使用工具来洞悉CF中的N2O的洞察力 航空公司可能会有利于特定微生物社区成员的适应性。 目的：AIM 1将确定导致通用CF细菌产生N2O的条件和途径 真菌，AIM 2将利用这些微生物产生的高级同位素分析N2O 从法医上讲，AIM 3将把这些见解应用于 随着肺功能下降，在此是否受到特定类型的硝化体。 提供指导早期翻译方法来治疗CF感染所需的关键知识 Airways以及建立工具，可以更多地应用于其他营养不良研究。