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.

项目摘要/摘要 拟议的研究计划的长期目标是提供对复杂对话的见解 人类与正常有益微生物群之间的发展。最近的研究表明有很强的 该对话的成功维护与终身健康之间的相关性。但是，无法获得 哺乳动物中殖民组织及其微生物的高度多样性的深度研究 人类肠道的持续殖民化极具挑战性。当面对如此复杂的时候 现象，生物学家经常转向更简单的模型系统，以提供对进化保守的见解 特征并揭示基本原则。破译了依据的细胞和分子机制 细菌与粘膜上皮的顶端表面的慢性缔合，拟议的程序利用了 特异性的二元共生，纤维杆菌fischeri及其鱿鱼宿主Euprymna scolopes。这 离散的，实验性的协会已被研究了将近三十年，作为该模型 革兰氏阴性细菌对粘膜的长期定植。就像在人类中一样 从每一代开始重新开始，并促进双方的持续健康。共生可以是 用带有定义突变的V. fischeri菌株发起的 生活。可以使用共聚焦显微镜直接对该协会进行成像，这为难得的机会提供了 定义具有较高的时间和空间分辨率，在终身的互惠分子和遗传对话中定义 有益关联。该项目结合了两个共同案件的专业知识，每一个都有经验 共生伙伴之一的生物学，并具有三个合作者的其他分析专业知识。一起， 他们将新方法和技术介绍给宿主微型互动的研究，包括：倒数 共生性共生对双方伴侣的影响的表观分析；杂交 - 链反应，荧光内 原位杂交（HCR-FISH），可以在宿主和共生细胞中可视化稀有转录本 定植组织；纳米弦技术，一种同时分析数十个到数百个的新方法 目标成绩单；并且，高效RNASEQ，从而产生强大的转录库 作为10 ng总RNA（约100,000个细菌）。要解决的具体目标是：（i）确定每日如何 代谢节奏在宿主和其微生物组之间进行了协调。 （ii）发现共生体是如何 保持生长和持久性的节奏循环； （iii）定义表观遗传修饰在 有益共生的发展和维护。对人类微生物组的了解 它的婴儿期，而这个边界领域目前处于建筑范式的阶段。在这种情况下，作为 鱿鱼频率系统过去具有，当前研究的结果将揭示基本原理 通过有益和致病性微生物来控制持续定殖。