Community-derived zinc metal regulation from monolayer to biofilm.

从单层到生物膜的群落衍生锌金属调节。

• 批准号: 10462373
• 负责人: Felix Steven Alfonso
• 金额: $ 6.76万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10462373
• 关键词: Affect; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Bacteria; Behavior; Binding; Biomedical Engineering; Biophysics; Birth; Cell Survival; Cells; Cellular Assay; Chemicals; Communication; Communities; Complex; Consumption; Cues; Data; Dependence; Development; Diet; Disease; Ecosystem; Environment; Escherichia coli; Excision; Fluorescence; Food; Future; Gene Expression; Genes; Gram-Negative Bacteria; Health; Home; Homeostasis; Human; Immune system; Individual; Infection; Intestines; Invaded; Knowledge; Life; Measures; Mediating; Mentors; Metals; Microbe; Microbial Biofilms; Microfluidic Microchips; Microfluidics; Micronutrients; Mission; Modeling; Monitor; National Institute of General Medical Sciences; Neighborhoods; Nutrient; Organism; Outcome; Pathway interactions; Play; Predictive Factor; Process; Proteins; Proxy; Public Health; Pump; Recording of previous events; Regulation; Research; Research Personnel; Role; Scheme; Side; Signal Transduction; Signaling Molecule; Social Network; Spectrum Analysis; System; Techniques; Testing; Time; Training; Transition Elements; Virulence; Virulent; Work; Zinc; antimicrobial; bacterial community; cell community; chemical genetics; chemical reaction; combat; design; efflux pump; environmental change; environmental stressor; fitness; genetic manipulation; genetically modified cells; gut microbiome; image processing; imaging platform; innovation; member; metal poisoning; microbial; microbial community; microbiome; monolayer; novel therapeutic intervention; opportunistic pathogen; optogenetics; pathogen; pathogenic microbe; periplasm; quorum sensing; recruit; response; side effect; single molecule; uptake

Project Summary and Abstract In humans, the gut is home to the most extensive set of diverse bacteria actively working together to break down nutrients for consumption, defend against pathogens, and train the immune system, as well as actively communicating with the host cells to optimize their survival. The gut microbiome formed shortly after birth changes over time in response to the diet and overall health of the host. When a pathogen invades the gut and adversely affects the host’s health, it is treated with antibiotics. However, the treatment has the side effect of indiscriminately altering the gut microbiome, leaving the host even more vulnerable to a future infection. Communities of bacterial cells maintain a state of homeostasis by actively communicating with each other and the host. This signaling system has the potential to serve as an innovative approach to treat virulent pathogens by recruiting the microbiome’s own defense system. However, it is unclear what metabolites serve as a signaling molecule to coordinate behavior. Transition metals play significant roles as micronutrients necessary to carry out complex chemical reactions required to sustain life. Consequently, their concentrations inside the cells are tightly regulated. This study focuses on zinc metal homeostasis due to its vital role in catalytic, structural, and regulatory functions in Escherichia coli, a model Gram-negative bacterium and a common bacterium in the environment, foods, and intestines. The overall objective of the proposed work is to determine whether zinc can act as a chemical cue to coordinate behavior in a community of cells in the context of metal homeostasis. My central hypothesis is that zinc acting as a signaling molecule can influence the cell’s neighborhood gene expression state to account for a changing environment in which the micronutrient is in low supply, excess, or used as a form of attack by a pathogen or the immune system. The hypothesis will be tested using combined approaches of microfluidics devices, chemical/genetic manipulations, optogenetics, single-molecule spectroscopy, and bulk biophysical/biomolecular/cellular assays. The proposed research has two specific aims: 1) Define the coordination of uptake and efflux capabilities among individual cells in a community as a function of zinc exposure. 2) Define the relation of periplasmic zinc concentration changes among individual cells in a community upon perturbation of their metal homeostasis. The applicant will be advised by a mentoring team that includes a chemist with expertise in single-molecule spectroscopy of bacterial metal uptake/efflux pumps, a biomedical engineer with expertise in microfluidic systems, and a microbiologist with expertise in bacterial metal homeostasis. The broader impact of this research is the creation of a quantitative model to describe how zinc metal homeostasis is achieved at the community level and delineate the role of the individual cells in a colony in facilitating homeostasis. The significance of this work is the creation of fundamental knowledge for help designing new innovative antimicrobial therapy that utilizes metal homeostasis. .

项目概要和摘要 在人类中，肠道是最广泛的各种细菌的家园，它们积极地共同努力， 减少营养素的消耗，抵御病原体，训练免疫系统，以及积极 与宿主细胞沟通以优化它们的存活。肠道微生物组在出生后不久形成 随着时间的推移而变化，以响应宿主的饮食和整体健康。当病原体侵入肠道， 对宿主的健康产生不利影响，则用抗生素治疗。然而，这种治疗有副作用， 不加区别地改变肠道微生物组，使宿主更容易受到未来的感染。 细菌细胞群落通过积极地相互交流来维持稳态， 主持人这种信号系统有可能作为一种创新的方法来治疗致命的病原体 通过招募微生物自身的防御系统。然而，目前还不清楚什么代谢物作为一种代谢物。 信号分子来协调行为。过渡金属作为人体必需的微量营养素， 进行维持生命所需的复杂化学反应。因此，它们在 细胞受到严格控制。这项研究的重点是锌金属稳态，因为它在催化， 大肠杆菌，一种革兰氏阴性菌模型和一种常见的 环境、食物和肠道中的细菌。拟议工作的总体目标是确定 锌是否可以作为一种化学线索，在金属环境中协调细胞群落的行为， 体内平衡我的中心假设是锌作为一种信号分子可以影响细胞的 邻近基因表达状态，以解释其中微量营养素处于低水平的变化环境。 病原体或免疫系统的供应、过量或用作攻击的形式。假设将被检验 使用微流体装置，化学/遗传操作，光遗传学， 单分子光谱和大量生物物理/生物分子/细胞测定。拟议的研究已 两个具体的目标：1）定义在一个特定的细胞中单个细胞之间的吸收和流出能力的协调， 作为锌暴露的函数。2)确定细胞周质锌浓度变化的关系 在一个群体中的单个细胞中，它们的金属稳态受到干扰。申请人将被 由一个指导团队提供建议，该团队包括一名具有单分子光谱学专业知识的化学家， 细菌金属吸收/外排泵，具有微流体系统专业知识的生物医学工程师，以及 在细菌金属稳态方面有专长的微生物学家这项研究更广泛的影响是创造了 一个定量模型来描述锌金属稳态是如何在社区水平上实现的， 描述了群体中单个细胞在促进体内平衡中的作用。这项工作的意义在于 创造基础知识，帮助设计新的创新抗菌治疗， 金属稳态 .