In vivo molecular analysis of probiotic impact on a defined microbiota and host

益生菌对特定微生物群和宿主影响的体内分子分析

• 批准号: 7896901
• 负责人: JUSTIN L SONNENBURG
• 金额: $ 5.4万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7896901
• 关键词: Address; Affect; Archaea; Arts; Bacteria; Bacteroides thetaiotaomicron; Bacteroidetes; Bifidobacterium; Biological Markers; Biology; Cecum; Cell Count; Cells; Clinical Research; Communities; Complex; Consumption; Country; Coupled; Cues; Development; Diet; Digestive System Disorders; Disease; Distal; Ecosystem; Electron Microscopy; Ensure; Environment; Epithelial; Epithelium; Eubacterium; Eukaryota; Evolution; Exposure to; Faculty; Fatty acid glycerol esters; Fermentation; Food; Fruit; Fucosyltransferase; Gastrointestinal tract structure; Genes; Genome; Glycobiology; Gnotobiotic; Grant; Harvest; Health; Homo sapiens; Human; Human Genome; Immune system; Indigenous; Individual; Inflammatory Bowel Diseases; Ingestion; Intestines; Inulin; Investigation; Lactic acid; Lactobacillus; Lasers; Life; Mammals; Mass Spectrum Analysis; Measures; Metabolic; Metabolic syndrome; Metabolism; Microbe; Modeling; Molecular; Molecular Analysis; Mucins; Mucous Membrane; Mucous body substance; Mus; Nutrient; Play; Polysaccharides; Positioning Attribute; Probiotics; Refrigeration; Research; Research Design; Role; Sanitation; Science; Societies; Source; Students; Symbiosis; Technology; Time; Training; Transmission Electron Microscopy; Universities; Washington; base; behavior influence; falls; foodborne; fortification; functional genomics; genome sequencing; glycosylation; gut microbiota; improved; in vivo; insight; lactic acid bacteria; member; microbial; microbiome; microorganism interaction; mouse model; response; trait

Ten to 100 trillion microbes inhabit our gastrointestinal tract (Gl) and have co-evolved with us to form an optimized symbiosis. Reduced occurrence of food-borne bacteria and increased sanitation in westernized countries over the past century has resulted in our Gl tract and resident microbiota being deprived exposure to lactic acid bacteria common in the diets of our ancestors. This proposal employs a gnotobiotic mouse model host to assess the impact of lactic-acid-producing probiotic bacterial species on (i) a model human microbiota composed of sequenced representatives of the two dominant bacterial divisions in our intestinal ecosystem, Bacteroides thetaiotaomicron (Bacteroidetes division) and Eubacterium rectale (Firmicutes division), and (ii) on the host. Bifidobacterium longum and Lactobacillus case/will represent the two common genera of probiotic bacteria. Aim 1 defines the impact of each probiotic species on the two-component model microbiota in the ceca of gnotobiotic mice. I will use whole genome transcriptional profiling of B. theta, E. rectale, and the probiotic species recovered from mouse ceca, transmission electron microscopy to assess bacterial localization, and mass-spectrometry of cecal nutrients. Aim 2 characterizes host epithelial responses and mucin remodeling induced by probiotics using transcriptional profiling of laser-capturedcecal epithelium. Mass-spectrometric analysis of cecal mucin glycosylation will provide a structural view of how glycans, which are utilized by microbes for nutrients and tethering, are modified as a function of probiotic exposure. Aim 3 assesses the impact of defined perturbations in the cecal nutrient environment on probiotic- symbiont-host interactions using (i) a defined inulin-rich diet, and (ii) a fucosyltransferase-deficient mouse that has altered cecal mucus composition. I have extensive training in host-microbial interactions, gnotobiotics, functional genomics, and glycobiology. The Washington University Center for Genome Sciences is a dynamic research environment that combines faculty and students with diverse expertise with state of the art technology, ensuring the support required as I transition into an independent faculty position. Relevance: This grant investigates the mechanisms by which probiotic bacteria affect the microbiota and Gl tract biology: given their wide use, these studies should provide a conceptual framework and biomarkers for hypothesis-based clinical studies designed to measure the impact of probiotics in healthy individuals and those with diseases inside and outside of the gut.

十到100万亿微生物居住在我们的胃肠道（GL），并与我们共同发展以形成 优化的共生。粮食传播细菌的发生减少和西方化的卫生 过去一个世纪的国家导致我们的GL区和居民微生物群被剥夺了暴露 在我们祖先的饮食中常见的乳酸细菌。该提案采用gnotobiotic鼠标 模型宿主评估乳酸产生益生菌细菌物种对（i）人类模型的影响 菌群由我们肠道中两个主要细菌分裂的测序代表组成 生态系统，细菌性thetaiotaomicron（杆菌分区）和菌群直肠（Firmicutes） 部门）和（ii）主机。双歧杆菌长杆菌和乳酸杆菌案例/将代表这两个常见 益生菌的属。 AIM 1定义了每种益生菌对两个组件的影响 在Gnotobiotic小鼠的CECA中模拟菌群。我将使用B. theta的整个基因组转录分析， E.直肠和从小鼠CECA回收的益生菌物种，传输电子显微镜至 评估细菌定位和盲肠营养素的质谱法。 AIM 2表征主机上皮 益生菌使用激光摄取的转录分析引起的反应和粘蛋白重塑 上皮。盲肠粘蛋白糖基化的质谱分析将提供结构性的视图 通过微生物用于营养和束缚的聚糖，被修饰为益生菌的函数 接触。 AIM 3评估定义的扰动在盲肠营养环境中对益生菌的影响 使用（i）定义的富含菊粉的饮食和（ii）岩藻糖基转移酶缺陷型小鼠的共生体 - 宿主相互作用 这改变了盲肠粘液组成。我对宿主微生物相互作用进行了广泛的培训， gnotobiotics，功能基因组学和糖生物学。华盛顿大学基因组中心 科学是一种充满活力的研究环境，将教职员工和具有不同专业知识的学生与 最先进的技术，确保当我过渡到独立教师职位时所需的支持。 相关性：该赠款研究了益生菌细菌影响菌群和GL的机制 研究生物学：鉴于它们的广泛使用，这些研究应为概念框架和生物标志物提供 基于假设的临床研究旨在衡量益生菌对健康个体的影响 肠道内外患有疾病的人。