Spatially Organized Bacterial Interaction Networks in the Gut Microbiota

肠道微生物群中空间组织的细菌相互作用网络

• 批准号: 10715436
• 负责人: Michael L Patnode
• 金额: $ 37.86万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-20 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10715436
• 关键词: Adhesions; Adult; Bacteria; Bacterial Adhesion; Bacterial Genes; Bacteroidetes; Binding; Cell Wall; Cells; Communicable Diseases; Exclusion; Family; Food; Generations; Gnotobiotic; Heterogeneity; Immune System Diseases; Intestinal Content; Intestines; Label; Libraries; Library Administration; Maps; Measures; Metabolic Diseases; Microbe; Mus; Mutagenesis; Mutation; Mutation Analysis; Oral Administration; Phenotype; Plants; Polysaccharides; Positioning Attribute; Process; Proteins; Site; Specificity; Structure; Technology; Time; dietary; gut microbes; gut microbiota; in vivo; magnetic beads; member; microbial; microbial community; particle; prevent

SUMMARY Each adult human harbors hundreds of bacterial species in their intestine. However, the networks of microbe- microbe interactions that underly the stable co-existence of resident species, and exclude additional species, are not well defined. The intestinal lumen is a turbulent, semi-fluid landscape where microbial cells and dietary plant cell wall fragments are distributed with high heterogeneity, and redistributed on the time scale of seconds. We propose that bacteria selectively adhere to dietary particles in the gut lumen and that interactions with their co-adherent microbial partners dictate whether they persist. We created multiplex libraries of artificial food particles (consisting of glycan-coated magnetic beads) to measure gut bacterial adhesion in vivo, and discovered that many members of the phylum Bacteroidetes adhere to dietary glycan particles in a strain- specific and glycan-specific manner. We will first identify families of adhesion proteins required for these binding phenotypes using transposon mutagenesis and insertion site sequencing. Next, we will identify networks of interacting strains that co-adhere to dietary particles. Using orally administered libraries of fluorescently labeled beads, we will map co-adhesion networks in gnotobiotic mice colonized with strains that have evolved together in a single donor host. Finally, we will establish a mutational selection strategy that permits the simultaneous generation of different binding specificities in genetically intractable gut microbes. Analysis of these mutations will reveal the potential origins of adhesion-dependent interspecies relationships. These studies will shed light on the poorly defined spatial structure of the gut microbiota. The technologies we develop in this proposal hold promise as a means to intentionally position the members of microbial communities in physical configurations that prevent or ameliorate metabolic, immunologic, and infectious diseases.

概括 每个成年人的肠道中都有数百种细菌。但是，微生物网络 居民物种稳定共存的微生物相互作用，并排除其他物种， 定义不当。肠腔是一种湍流的半流体景观，其中微生物细胞和饮食 植物细胞壁碎片分布高异质性，并在秒的时间尺度上重新分布。 我们建议细菌选择性地遵守肠道中的饮食颗粒，并与其相互作用 共遵守的微生物伴侣决定是否坚持。我们创建了人造食品的多重库 颗粒（由聚糖涂层的磁珠组成），用于在体内测量肠道细菌粘附，并且 发现菌群的许多成员都粘在菌株中的饮食聚糖颗粒 特定和特定于聚糖的方式。我们将首先确定这些粘附蛋白所需的家族 使用转座子诱变和插入位点测序的结合表型。接下来，我们将确定 相互作用的菌株网络，与饮食颗粒共同添加。使用口头管理的库 荧光标记为珠子，我们将在gnotobiotic小鼠中绘制与菌株定居的gnotobiotic小鼠的共粘附网络 在单个捐助者主持人中一起进化。最后，我们将建立一种突变选择策略 允许在遗传上棘手的肠道微生物中同时产生不同的结合特异性。 对这些突变的分析将揭示粘附依赖性间隙关系的潜在起源。 这些研究将阐明肠道菌群的定义较差的空间结构。我们的技术 在此提案中发展有望作为有意定位微生物成员的一种手段 预防或改善代谢，免疫和感染力的物理配置的社区 疾病。