Community-derived zinc metal regulation from monolayer to biofilm.

项目摘要和摘要 在人类中，肠道是最广泛的一组不同细菌的家园，它们积极地共同工作，以打破 摄取营养物质，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生存。肠道微生物群在出生后不久就形成了 随着时间的推移而变化，以回应主人的饮食和整体健康状况。当病原体侵入肠道并 会对宿主的健康造成不利影响，需要使用抗生素进行治疗。然而，这种治疗的副作用是 不加区别地改变肠道微生物群，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和 主持人。这种信号系统有可能成为治疗致病毒力的创新方法。 通过招募微生物组自身的防御系统。然而，目前还不清楚哪些代谢物起到了 信号分子来协调行为。过渡金属作为必需的微量营养素发挥着重要作用 进行维持生命所需的复杂化学反应。因此，它们在体内的浓度 细胞受到严格的监管。本研究的重点是锌金属的动态平衡，因为它在催化， 作为革兰氏阴性杆菌的模式菌和常见的 环境、食物和肠道中的细菌。拟议工作的总体目标是确定 在金属环境中，锌是否可以作为一种化学线索来协调细胞群落的行为 动态平衡。我的中心假设是，锌作为信号分子可以影响细胞的 邻域基因表达状态，以解释微营养素处于低水平的环境变化 供过于求由病原体或免疫系统提供、过量或用作攻击的一种形式这一假设将得到检验。 使用微流控设备、化学/遗传操作、光遗传学、 单分子光谱学和整体生物物理/生物分子/细胞分析。拟议中的研究已经 两个特定的目标：1)定义细胞间摄取和排出能力的协调 社区作为锌暴露的函数。2)明确周质锌浓度变化的关系 在一个群落中的单个细胞之间，当它们的金属稳态被扰动时。申请者将是 由一个指导团队提供建议，该团队包括一名在单分子光谱学方面具有专业知识的化学家 细菌金属摄取/外排泵，具有微流控系统专业知识的生物医学工程师，以及 在细菌金属动态平衡方面有专长的微生物学家。这项研究的更广泛的影响是创造 描述锌金属动态平衡是如何在群落水平上实现的，以及 描述菌落中单个细胞在促进动态平衡中的作用。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疗法，利用 金属动态平衡。 。

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. .