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. .

项目摘要和摘要 在人类中，肠道是最广泛的潜水细菌的家园 减少养分供食用，防御病原体和训练免疫系统，并积极地 与宿主细胞进行通信以优化其生存。出生后不久形成的肠道微生物组 随着时间的流逝，宿主的饮食和整体健康状况会随着时间的流逝而变化。当病原体侵入肠道时 不利地影响宿主的健康，并用抗生素治疗。但是，治疗具有 不加选择地改变肠道微生物组，使宿主更容易受到未来感染的影响。 细菌细胞的群落通过彼此积极交流，维持稳态的状态 主人。该信号系统有可能用作治疗有毒病原体的创新方法 通过招募微生物组的防御系统。但是，目前尚不清楚哪些代谢物用作 信号分子以协调行为。过渡金属在必要的微量营养素中起着重要作用 进行维持生命所需的复杂化学反应。因此，他们的浓度 细胞受到严格调节。这项研究的重点是锌金属稳态，因为它在催化中的重要作用， 大肠杆菌的结构和调节功能，模型革兰氏阴性细菌和常见 环境中的细菌，食品和肠道。拟议工作的总体目标是确定 在金属背景下，锌是否可以充当化学提示以协调细胞社区的行为 稳态。我的中心假设是，充当信号分子的锌会影响细胞的 邻域基因表达状态可以解释微量营养素不断变化的环境 供应，超过或用作病原体或免疫系统的攻击形式。该假设将进行检验 使用微流体设备，化学/遗传操纵，光遗传学的联合方法 单分子光谱和大量生物物理/生物分子/细胞测定。拟议的研究已经 两个具体的目的：1）定义一个单个细胞中单个细胞中吸收和外排能力的协调 社区是锌暴露的功能。 2）定义周质锌浓度变化的关系 在社区中的单个细胞中，由于其金属稳态扰动。申请人将 由一个心理团队的建议，其中包括一名具有单分子光谱专业知识的化学家 细菌金属摄取/外排泵，具有微流体系统专业知识的生物医学工程师和A 具有细菌金属稳态专业知识的微生物学家。这项研究的更广泛的影响是创作 定量模型，以描述在社区层面上如何实现锌金属稳态和 描述单个细胞在菌落支持稳态中的作用。这项工作的意义是 创建基本知识来帮助设计使用新的创新抗菌疗法 金属稳态。 。