Spatial Organization of the Oral Microbiome

口腔微生物组的空间组织

基本信息

批准号：
10803528
负责人：
Jessica Leigh Mark Welch
金额：
$ 78万
依托单位：
ADA FORSYTH INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-03 至 2028-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10803528
关键词：
Actinomyces Address Affect Animals Architecture Bacteria Biological Models Biology Cells Characteristics Communities Complex Complex Mixtures Corynebacterium DNA Sequence Analysis Data Dental Plaque Development Diameter Disease Dyes Ecology Environment Fluorescent in Situ Hybridization Foundations Fusobacterium Generations Goals Grant Growth Health Human Image In Vitro Individual Industrialization Label Learning Measures Metabolism Metagenomics Methods Microbial Biofilms Microdissection Modeling Neighborhoods Neisseria Nitrates Nitric Oxide Oral Oral cavity Oral health Persons Physiology Play Population Positioning Attribute Prevotella Research Resolution Role Saliva Salivary Salivary Glands Shotguns Site Stains Streptococcus Structure Structure-Activity Relationship Supplementation Surface System Taxon Tissues Tongue Vasodilation Veillonella Whole-Genome Shotgun Sequencing Work bacterial community blood pressure control cardiometabolism combinatorial community organizations dietary nitrate host microbiome imaging system improved in vitro Model in vivo live cell imaging member meter microbial microbial community microbiome microbiota microbiota transplantation millimeter novel therapeutics oral biofilm oral microbial community oral microbiome probiotic therapy response spatial relationship spectrograph tongue dorsum volunteer whole genome

项目摘要

TITLE: Spatial Organization of the Oral Microbiome PI: Jessica Mark Welch Project Summary/Abstract Bacteria colonizing the human mouth play important roles not only in oral health, but in systemic health and disease. This proposal addresses the key unsolved question of how spatial organization of the oral microbiota is connected to its function at the micron scale. Our previous work pioneered the imaging of oral microbiome biogeography by developing a Combinatorial Labeling And Spectral Imaging - Fluorescence in situ Hybridization (CLASI-FISH) strategy to analyze the micron-scale architecture of microbial communities at a systems level. In this project we build on these findings to investigate structure-function relationships, assess complexity of consortia at micrometer scales within single human hosts, and develop multiplexed live cell imaging to study the dynamics of multispecies in vitro biofilms at the single-cell level. In Aim 1 we investigate how a key parameter of host-microbiome interaction, salivary nitrate concentration, affects the structure of the bacterial biofilm on the tongue dorsum. The nitrate-reducing capability of bacteria on the tongue plays a major role in the generation of nitric oxide, which impacts vasodilation and control of blood pressure. In turn, the secretion of nitrate into saliva by salivary glands affects the metabolism and composition of the tongue microbiota. Using dietary nitrate supplementation of volunteers, we will quantify the spatial organization of nitrate-metabolizing taxa in tongue biofilm consortia that developed in high and low salivary nitrate concentrations, and we will use whole-genome shotgun sequence data to determine which species and strains are responsive to nitrate. In Aim 2 we address an almost entirely unexplored question: in micron-scale neighborhoods in the oral microbiome, does each species function as a clone or as a complex mixture of strains? The answer to this question is important for learning to manipulate and modulate the oral microbiome and determining whether, for purposes such as microbiome transplants or building an in vitro biofilm model, a single strain is an adequate representative of the community. We will address this question by microdissection of single tongue dorsum consortia 100-200 micrometers in diameter, followed by whole-genome shotgun sequencing and analysis to assess whether natural oral consortia are made up of clones or complex populations. Live imaging is a powerful method of analysis of taxon-taxon interactions, but multiplexed live imaging of bacteria with single-cell resolution has been almost entirely absent from studies of the oral microbiome. The goal of Aim 3 is to develop a live-imaging system, using vital dyes, to differentiate up to 6 taxa within live simplified communities. We will use this system to investigate the dynamics of taxon-taxon interactions in consortia representing components of dental plaque and the tongue biofilm. Our goal is to learn how members of the oral microbiome respond to one another under controlled conditions, and to evaluate whether these in vitro systems develop spatial organization resembling those of natural communities.

标题：口腔微生物组的空间组织 PI：杰西卡·马克·韦尔奇项目概要/摘要人类口腔中的细菌不仅在口腔健康中发挥着重要作用，而且在全身健康中发挥着重要作用和疾病。该提案解决了尚未解决的关键问题：口语的空间组织如何微生物群与其功能在微米尺度上相关。我们之前的工作开创了口腔成像通过开发组合标记和光谱成像 - 原位荧光来进行微生物组生物地理学杂交 (CLASI-FISH) 策略可在一定范围内分析微生物群落的微米级结构系统级别。在这个项目中，我们以这些发现为基础来研究结构-功能关系，评估单个人类宿主内微米级联合体的复杂性，并开发多重活细胞成像在单细胞水平上研究多物种体外生物膜的动态。在目标 1 中，我们研究了宿主-微生物组相互作用的关键参数——唾液硝酸盐如何变化。浓度，影响舌背上细菌生物膜的结构。硝酸盐还原剂舌头上细菌的能力在一氧化氮的产生中起着重要作用，一氧化氮会影响血管舒张和血压控制。反过来，唾液腺将硝酸盐分泌到唾液中会影响舌头微生物群的新陈代谢和组成。使用志愿者的饮食补充硝酸盐，我们将量化开发的舌头生物膜联盟中硝酸盐代谢类群的空间组织在高和低唾液硝酸盐浓度下，我们将使用全基因组鸟枪序列数据来确定哪些物种和菌株对硝酸盐有反应。在目标 2 中，我们解决了一个几乎完全未被探索的问题：在口腔中的微米级邻域中微生物组，每个物种是作为克隆还是作为菌株的复杂混合物发挥作用？这个问题的答案这个问题对于学习操纵和调节口腔微生物组并确定是否，为了微生物组移植或建立体外生物膜模型等目的，单一菌株是足够的社区代表。我们将通过单舌显微解剖来解决这个问题背部联合体直径100-200微米，然后进行全基因组鸟枪测序和分析以评估自然口腔群落是否由克隆或复杂群体组成。实时成像是分析分类单元-分类单元相互作用的强大方法，但多重实时成像口腔微生物组的研究几乎完全不存在单细胞分辨率的细菌。这目标 3 的目标是开发一个实时成像系统，使用活性染料来区分活体中多达 6 个类群简化的社区。我们将使用该系统来研究分类单元-分类单元相互作用的动态代表牙菌斑和舌头生物膜成分的联合体。我们的目标是了解会员如何口腔微生物组在受控条件下相互反应，并评估这些微生物是否在体外系统发展出类似于自然群落的空间组织。