Vibrio colonization determinants in symbiosis

共生中弧菌定植的决定因素

• 批准号: 10797149
• 负责人: MARGARET J MC FALL-NGAI
• 金额: $ 20万
• 依托单位: CARNEGIE INSTITUTION OF WASHINGTON, D.C.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10797149
• 关键词: Address; Adopted; Animals; Apical; Bacteria; Biological Models; Biology; Breeding; Cells; Chronic; Communication; Complex; Confocal Microscopy; DNA Methylation; Development; Developmental Biology; Epigenetic Process; Epithelium; Euprymna scolopes; Fluorescent in Situ Hybridization; Fostering; Generations; Genetic; Genetic Transcription; Genomics; Goals; Gram-Negative Bacteria; Growth; Health; Human; Human Microbiome; Image; Libraries; Life; Light; Maintenance; Mammals; Metabolic; Methods; Modeling; Modification; Molecular; Mucous Membrane; Mutation; Periodicity; Physiological; RNA; Reaction; Regulation; Research; Resolution; Role; Scientist; Squid; Surface; Symbiosis; Techniques; Technology; Tissues; Transcript; V. fischeri-squid system; Vibrio; Vibrio fischeri; Visualization; Work; bacterial genetics; beneficial microorganism; epigenomics; experience; frontier; host-microbe interactions; infancy; innovation; insight; metabolomics; microbiome; microbiota; nano-string; new technology; novel; novel strategies; pathogenic microbe; programs; symbiont; transcriptome sequencing; transcriptomics

Project Summary/Abstract The long-term goals of the proposed research program are to provide insight into the complex dialogue that has evolved between humans and their normal beneficial microbiota. Recent research has indicated a strong correlation between the successful maintenance of this dialog and life-long health. However, the inaccessibility of colonized tissues in mammals and the high diversity of their microbiomes render an in-depth study of persistent colonization of the human gut track extremely challenging. When faced with such complex phenomena, biologists often turn to simpler model systems to provide insights into evolutionarily conserved features and reveal basic principles. To decipher the cellular and molecular mechanisms underlying the chronic association of bacteria with apical surfaces of mucosal epithelia, the proposed program exploits the specific, binary symbiosis between the bacterium Vibrio fischeri and its squid host, Euprymna scolopes. This discrete, experimentally tractable, association has been studied for almost three decades as a model for the long-term colonization of mucosa by Gram-negative bacteria. As in humans, the squid-vibrio association begins anew each generation, and fosters the continuing health of both partners. The symbiosis can be initiated with V. fischeri strains with defined mutations, and the host can be bred and maintained for its entire life. The association can be directly imaged using confocal microscopy, which offers the rare opportunity to define, with high temporal and spatial resolution, the reciprocal molecular and genetic dialogue in a life-long beneficial association. This project combines the expertise of the two co-PIs, each with experience in the biology of one of the symbiotic partners, with additional analytical expertise of three collaborators. Together, they introduce new approaches and technology to the study of host-microbe interactions, including: reciprocal epigenomic analyses of the effect of symbiosis on both partners; hybridization-chain-reaction, fluorescent in situ hybridization (HCR-FISH), which enables visualization of rare transcripts in both host and symbiont cells in colonized tissues; NanoString Technology, a new method for simultaneous analysis of dozens to hundreds of targeted transcripts; and, high-efficiency RNAseq, which produces robust transcriptional libraries from as little as 10 ng total RNA (~100,000 bacteria). Specific aims to be addressed are: (i) determining how a daily metabolic rhythm is coordinated between a host and its microbiome; (ii) discovering how the symbionts maintain a rhythmic cycle of growth and persistence; and, (iii) defining the role of epigenetic modifications in the development and maintenance of a beneficial symbiosis. An understanding of the human microbiome is in its infancy, and this frontier field is currently at the stage of building paradigms. Within this context, as the squid-vibrio system has in the past, the results of the current study will shed light upon fundamental principles that govern persistent colonization by both beneficial and pathogenic microbes.

项目概要/摘要 拟议研究计划的长期目标是深入了解复杂的对话， 是在人类与其正常有益微生物群之间进化而来的。最近的研究表明，强烈的 成功维持这种对话与终生健康之间的相关性。然而，交通不便 哺乳动物的定植组织及其微生物组的高度多样性使得深入研究 人类肠道轨迹的持续定植极具挑战性。当面对如此复杂的情况时 对于现象，生物学家经常转向更简单的模型系统来提供对进化保守性的见解 特征并揭示基本原理。破译潜在的细胞和分子机制 细菌与粘膜上皮顶端表面的慢性关联，拟议的计划利用了 费氏弧菌及其宿主鱿鱼 Euprymna scolopes 之间存在特定的二元共生关系。这 离散的、实验上易于处理的关联作为一个模型已经被研究了近三十年。 革兰氏阴性菌在粘膜上长期定植。与人类一样，鱿鱼-弧菌协会 每一代人都重新开始，并促进双方的持续健康。共生可以是 由具有明确突变的 V. fischeri 菌株开始，并且宿主可以在整个过程中进行繁殖和维持 生活。该关联可以使用共焦显微镜直接成像，这提供了难得的机会 以高时间和空间分辨率定义终生的相互分子和遗传对话 有益的关联。该项目结合了两位联合 PI 的专业知识，每人都拥有以下方面的经验： 共生伙伴之一的生物学，以及三名合作者的额外分析专业知识。一起， 他们引入了新的方法和技术来研究宿主-微生物相互作用，包括： 共生对双方影响的表观基因组分析；杂交链式反应，荧光 原位杂交 (HCR-FISH)，可实现宿主细胞和共生细胞中稀有转录本的可视化 定植组织； NanoString技术，一种同时分析数十至数百个的新方法 目标转录本；以及高效的 RNAseq，它可以用尽可能少的量生成强大的转录文库 为 10 ng 总 RNA（约 100,000 个细菌）。要解决的具体目标是： (i) 确定每天如何 代谢节律在宿主与其微生物组之间协调； (ii) 发现共生体如何 保持有节奏的生长和持久循环； (iii) 定义表观遗传修饰在 发展和维持有益的共生关系。对人类微生物组的了解在于 该前沿领域尚处于起步阶段，目前正处于构建范式的阶段。在此背景下，作为 鱿鱼弧菌系统过去已经存在，当前的研究结果将揭示基本原理 控制有益微生物和病原微生物的持续定植。