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

测试以下假设：微生物对 CF 免疫反应的能量劫持会在肺功能下降期间选择特定病原体 - Diversity Supplement

• 批准号: 10745232
• 负责人: Dianne K Newman
• 金额: $ 1.34万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-03 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10745232
• 关键词: Academia; Address; Adopted; Adult; American; Antibiotics; Articulation; Automobile Driving; Bacteria; Bacterial Physiology; Bacteriology; Biology; Biomedical Research; Biotechnology; Carbon; Career Choice; Cell Respiration; Chalk; Charge; Chronic; Communication; Complex; Creativeness; Cystic Fibrosis; Cystic Fibrosis sputum; Data; Development Plans; Earth science; Educational process of instructing; Energy-Generating Resources; Ensure; Environment; Epithelial Cells; Feedback; Floods; Forensic Medicine; Gases; Generations; Genetic; Goals; Government; Group Meetings; Growth; Guidelines; Habitats; Happiness; Human; Image; Immune response; Immune system; Individual; Industry; Internships; Isotopes; Knowledge; Leadership; Learning; Lung infections; Manuscripts; Measurable; Membrane; Microbe; Microbiology; Molecular Profiling; Mucous body substance; Nitric Oxide; Oxygen; Paper; Pathway interactions; Pilot Projects; Policies; Process; Production; Public Speaking; Publications; Pulmonary Cystic Fibrosis; Reproducibility; Research; Research Training; Respiratory Tract Infections; Role; Scientist; Societies; Soil; Source; Structure; Students; System; Technical Expertise; Testing; Time; Tobramycin; Training; Training Programs; United States National Institutes of Health; Work; Writing; career; combat; cystic fibrosis airway; cystic fibrosis infection; cystic fibrosis patients; denitrification; doctoral student; dysbiosis; experience; experimental study; fitness; fungus; host-microbe interactions; in situ imaging; insight; lung failure; meetings; member; microbial; microbial community; microbiota; neutrophil; opportunistic pathogen; oxidation; pathogen; pre-doctoral; programs; pulmonary function decline; responsible research conduct; skills; symposium; tool; 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 患有囊性纤维化（CF）的个体与气道的破坏性慢性微生物感染作斗争。 虽然CF患者通常是由其他无辜的细菌和真菌-其中许多来自 受到损害的CF免疫系统无法清除这些机会， 随后宿主和微生物之间的斗争最终导致肺部系统的衰竭。常规 抗生素不是很有效。一个广泛接受的观点是，为什么会出现这种情况， 生长缓慢，它们的细胞膜不足以吸收常用的抗生素， 妥布霉素。因此，如果我们寻求新的方法来治疗CF肺部感染，我们必须更好地了解如何 微生物在这里繁衍生息不同的证据表明，CF气道中的氧气是有限的， 与机会致病菌相关的微观尺度。尽管充满粘液的 呼吸道，在没有氧气的情况下，细菌和真菌来统治这个栖息地必须雇用 有氧呼吸的替代能源产生策略。他们是怎么做到的？能量流是 任何微生物群落的组织者，包括CF气道中发现的那些。众所周知， CF患者免疫系统产生的一氧化氮（NO）低于健康人， 个体事实上，上皮细胞在NO的产生中受到严重损害，然而， CF气道仍然可以产生NO;这种NO不足以杀死微生物，但有可能转化为 转化为那些能够脱氮的生物的能量来源。因此，我们试图检验假设， 通过反硝化途径对CF免疫应答的微生物能量劫持选择特异性 肺功能下降期间的病原体。为了验证这一假设，我们计划利用强大的同位素工具 从地球科学，允许在复杂的环境中的代谢物的来源，如CF 通过一个原子一个原子地测定，提供一个非侵入性的法医分子指纹。特别是， 我们将重点解释N2 O，一种CF痰和呼吸中脱氮途径的可测量产物 气体除了采用细菌生理学、遗传学和宿主微生物原位成像的方法外， 相互作用，我们将使用这些同位素工具来深入了解如何在CF中还原为N2 O的NO 气道可能有利于特定微生物群落成员的适应性。为此，我们提出了三个具体的建议。 目的：目的1将确定导致常见CF细菌产生N2 O的条件和途径， 真菌，目标2将利用先进的同位素分析来解剖这些微生物产生的N2 O， 法医推断其来源，目标3将这些见解应用于成人CF患者的试点研究，以确定 当肺功能下降时，特定类型的净化剂是否更受欢迎。实现这些目标将 提供指导CF感染的早期转化方法所需的关键知识 气道，以及建立可以更广泛地应用于其他生态失调研究的工具。