Engineering microbiomes and their molecular determinants for production of butyrate and secondary bile acids from resistant starch

利用抗性淀粉生产丁酸和次级胆汁酸的工程微生物组及其分子决定因素

• 批准号: 10441582
• 负责人: Thomas M Schmidt
• 金额: $ 37.04万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10441582
• 关键词: Age; Agonist; Anabolism; Anaerobic Bacteria; Attenuated; Bacteria; Bifidobacterium; Bile Acids; Binding; Biogenesis; Bioinformatics; Biological Models; Butyrates; Carbohydrates; Carbon; Cell Fraction; Cell Nucleus; Cell Respiration; Cells; Clinical; Communities; Consumption; Energy-Generating Resources; Engineering; Environment; Enzymes; Epithelial Cells; Excision; Exposure to; Fermentation; Fiber; Food Webs; Gene Expression; Germ-Free; Gnotobiotic; Goals; Gut Mucosa; Harvest; Human; Hydrogen; Hydrolase; Hydrolysis; In Vitro; Incidence; Inflammation; Internet; Measures; Mediating; Mesalamine; Metabolic; Metabolism; Microbe; Mitochondria; Modeling; Molecular; Molecular Probes; Mucous Membrane; Mus; Oxides; Oxygen; PPAR alpha; Patient-Focused Outcomes; Patients; Peroxisome Proliferator-Activated Receptors; Physiological; Potato; Production; Proteins; Proteomics; Receptor Activation; Resistance; Respiration; Ruminococcus; Scaffolding Protein; Severities; Starch; Succinate Dehydrogenase; Taxonomy; Testing; Therapeutic; Time; Tissues; Transplant Recipients; Volatile Fatty Acids; bile salts; cohort; cytochrome c oxidase; dietary supplements; gastrointestinal epithelium; graft vs host disease; gut bacteria; gut inflammation; gut microbes; gut microbiome; gut microbiota; hematopoietic cell transplantation; high throughput screening; in vivo; intestinal epithelium; microbial; microbial community; microbiome; microbiome analysis; mouse model; oxidation; particle; preservation; response; western diet

PROJECT SUMMARY/ABSTRACT – PROJECT 3 The production of butyrate from gut microbiomes results from interactions between microbes in anaerobic food webs. Identifying butyrogenic combinations of microbes, environmental conditions and fermentable fibers will underlie strategies for manipulating the microbiomes in BMT patients to provide therapeutic concentrations of butyrate. Hypotheses: 1) Maximal production of SCFAs from fermentable fibers requires the combination of a primary fiber degraders, secondary fermenters and hydrogen-consuming microbes. 2) The ratio of butyrate to total SCFAs is dependent on the taxonomic membership of communities, which is selected by the concentrations of H2, bile acids, pH and turnover time of the environment. Approaches: We will use covariation analysis of microbiomes from a healthy human cohort to identify butyrogenic combinations of fermentable fibers, physical conditions and microbes. We will test these predictions by assembling synthetic communities of the identified microbes and measuring butyrate production in vitro under various conditions, including the removal of hydrogen by hydrogenotrophic microbes. We will also select co-evolved butyrogenic communities from fecal inocula using multiple passages through media containing fermentable fiber as the primary carbon and energy source. Once transported into an intestinal epithelial cell, butyrate can be oxidized by mitochondria. It and can also stimulate mitochondrial biogenesis through Peroxisome Proliferator-Activated Receptors (PPARs) located on the nucleus. Understanding the interaction between a butyrogenic microbiome and epithelial cells will provide a mechanistic explanation of the temporal dynamics between butyrate production and host cell respiration. Hypothesis: In vitro and in vivo respiration and mitochondrial biogenesis in colonic epithelial cells will be stimulated by butyrate. Approaches: Respiration and PPAR activation will be measured in enteroids exposed to butyrate under varying environmental conditions. For in vivo estimates of respiration, mice at six weeks of age will be placed on a Western diet supplemented with a fermentable fiber(s) or accessible starch. GI tissues will be harvested at intervals up to 12 weeks and succinate dehydrogenase and cytochrome oxidase activities will be measured to quantify the capacity for cellular respiration. Cell respiration and mucosal O2 concentrations will be tracked in germ-free mice treated 5- aminosalicylic acid, a PPAR agonist, to distinguish between mitochondrial biosynthesis and butyrate oxidation. Mice colonized with synthetic communities of microbes to be used to test the impact of microbiomes of different butyrogenic capacity on respiration.

项目摘要/摘要 - 项目3 来自肠道微生物组的丁酸酯的产生是由于厌氧中微生物之间的相互作用而产生的 食物网。鉴定微生物，环境条件和可发酵的丁基组合 纤维将是操纵BMT患者微生物组的策略的基础 丁酸酯的浓度。假设：1）可发酵纤维的最大生产SCFA 需要将主要纤维降解器，次级发酵罐和氢的组合组合 微生物。 2）丁酸酯与总SCFA的比率取决于分类成员资格 社区，由H2，胆汁酸，pH和周转时间的浓度选择 环境。方法：我们将使用来自健康人的微生物组的协方差分析 鉴定可发酵纤维，物理条件和微生物的丁酚组合。我们 将通过组装确定的微生物的合成群落并测量来测试这些预测 丁酸酯在各种条件下在体外产生，包括 营养微生物。我们还将从粪便接种中选择共同进化的丁基群落 使用多个通过包含可发酵纤维作为主要碳和能量的培养基 来源。 丁酸丁酸酯一旦进入肠上皮细胞，就可以用线粒体氧化。它可以 还通过过氧化物体增殖物激活受体（PPAR）刺激线粒体生物发生 位于细胞核上。了解丁基微生物组和上皮之间的相互作用 细胞将对丁酸酯产生和 宿主细胞呼吸。假设：结肠中的体外和体内呼吸和线粒体生物发生 上皮细胞将被丁酸酯刺激。方法：呼吸和PPAR激活将是 在不同的环境条件下暴露于丁酸酯的肠to虫中测量。用于体内估计 在呼吸中，将六周大的小鼠放置在补充可发酵的西方饮食上 纤维或可进入的淀粉。胃肠道组织将以最长12周的间隔收获，并琥珀 将测量脱氢酶和细胞色素氧化物活性，以量化细胞的能力 呼吸。细胞呼吸和粘膜O2浓度将在5--处理的无菌小鼠中追踪 PPAR激动剂氨基酸，以区分线粒体生物合成和丁酸酯 氧化。用微生物的合成群落定殖的小鼠用于测试 不同丁基能力呼吸的微生物组。