Harnessing the gut microbiome to detect and quantify glycans

利用肠道微生物组来检测和量化聚糖

• 批准号: 10693304
• 负责人: Guy Edmund Townsend
• 金额: $ 39.4万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10693304
• 关键词: Affinity; Affinity Chromatography; Animals; Automobile Driving; Bacteroides; Bacteroides thetaiotaomicron; Binding Proteins; Biological; Biological Process; Carbohydrates; Complement; Complex; Complex Mixtures; Detection; Disease; Dose; Engineering; Environment; Genes; Goals; Harvest; Human; In Vitro; Individual; Intestinal Content; Intestines; Libraries; Life; Mass Spectrum Analysis; Measurement; Measures; Metabolic; Metabolism; Methods; Microbe; Modeling; Modification; Natural regeneration; Nuclear Magnetic Resonance; Nucleic Acids; Nutrient; Oligonucleotides; Organism; Output; Plants; Polymers; Polysaccharides; Proteins; Recombinants; Reporter; Reporting; Resources; Sampling; Sensitivity and Specificity; Source; Specificity; Standardization; Structure; Surveys; System; Therapeutic; biological preparation; clinically relevant; cost; detection method; detection platform; gut bacteria; gut microbes; gut microbiome; gut microbiota; health determinants; interest; member; microbial; microbiome composition; novel; predictive tools; response; therapeutic target; tool; tool development; Affinity; Affinity Chromatography; Animals; Automobile Driving; Bacteroides; Bacteroides thetaiotaomicron; Binding Proteins; Biological; Biological Process; Carbohydrates; Complement; Complex; Complex Mixtures; Detection; Disease; Dose; Engineering; Environment; Genes; Goals; Harvest; Human; In Vitro; Individual; Intestinal Content; Intestines; Libraries; Life; Mass Spectrum Analysis; Measurement; Measures; Metabolic; Metabolism; Methods; Microbe; Modeling; Modification; Natural regeneration; Nuclear Magnetic Resonance; Nucleic Acids; Nutrient; Oligonucleotides; Organism; Output; Plants; Polymers; Polysaccharides; Proteins; Recombinants; Reporter; Reporting; Resources; Sampling; Sensitivity and Specificity; Source; Specificity; Standardization; Structure; Surveys; System; Therapeutic; biological preparation; clinically relevant; cost; detection method; detection platform; gut bacteria; gut microbes; gut microbiome; gut microbiota; health determinants; interest; member; microbial; microbiome composition; novel; predictive tools; response; therapeutic target; tool; tool development

Project Summary Detecting various compositionally similar but structurally distinct glycans present in heterogenous mixtures has traditionally been challenging due to the limitations facing current glycomics approaches. These limitations have hindered our understanding of the availability and abundance of individual glycans in the mammalian gut environment, which is replete with diverse mixtures of microbial, mammalian, and plant-derived oligo- and polysaccharides. Moreover, the intestinal glycan composition is a primary driver of gut microbiome composition and metabolism, which represents an increasingly important human health determinant, and necessitates a deep understanding of the glycomic-microbial interface to identify important biological interactions and putatively develop glycan-derived therapeutics to target specific microbial activities. Therefore, new tools are necessary to detect and measure the relative abundance of individual glycan substrates present in the heterogenous mixtures prepared from biological sources such as mammalian intestinal contents. We have harnessed the glycan detection machinery employed by dominant members of the gut microbiota to detect, measure and isolate individual glycan substrates present in heterogenous mixtures extracted from the mammalian intestine. Herein, we demonstrate robust, specific, and scalable approaches by which engineered microbes report the presence of individual glycan substrates with incredible sensitivity. Furthermore, we demonstrate that this approach can accurately measure the abundance of individual glycans present in mixtures across wide linear ranges and that the specificity and sensitivity of these measurements can be tuned by modifying particular microbial glycan utilization genes. Finally, we demonstrate that microbially-encoded glycan-binding proteins can be used to isolate individual target glycans from mixtures for downstream compositional and structural determination. We propose to 1.) develop arrayed libraries of distinct gut microbial species, each engineered to report the presence of unique target glycans, 2.) develop a rapid glycan isolation pipeline to purify individual substrates of interest for downstream structural and functional characterization, and 3.) develop genetically modified microbial strains with enhanced sensitivity or target specificity. In addition to offering a rapid and inexpensive alternative to quantifying known glycans, we believe that further development of these tools will reveal the presence and abundance of previously undetectable glycans and dramatically enhance our understanding of the interactions between gut microbes and their mammalian hosts.

项目摘要 在异质混合物中发现各种成分相似但结构上不同的聚糖具有 传统上，由于当前的糖基质方法面临的局限性，这是充满挑战的。这些限制有 阻碍了我们对哺乳动物肠中个体聚糖的可用性和丰度的理解 环境，充满了微生物，哺乳动物和植物衍生的寡做和的多种混合物 多糖。此外，肠道聚糖成分是肠道微生物组组成的主要驱动力 和代谢，这代表了越来越重要的人类健康决定因素，因此需要深入 了解糖菌微生物界面，以识别重要的生物学相互作用和推测 开发基于聚糖的治疗剂，以靶向特定的微生物活性。因此，新工具是必要的 检测并测量异源混合物中存在的单个聚糖基质的相对丰度 由生物学来源（例如哺乳动物肠含量）制备。我们利用了聚糖 肠道微生物群主导成员使用的检测机制来检测，测量和分离 从哺乳动物肠中提取的异质混合物中存在的各个聚糖底物。在此处， 我们展示了强大的，特定和可扩展的方法，工程化的微生物报告了存在 具有令人难以置信的灵敏度的个体聚糖基质。此外，我们证明这种方法可以 准确地测量跨线性范围混合物中存在的单个聚糖的丰度， 这些测量值的特异性和灵敏度可以通过修改特定的微生物聚糖来调整 利用基因。最后，我们证明了微生物编码的聚糖结合蛋白可用于分离 来自混合物的单个靶向糖，用于下游组成和结构测定。我们建议 到1.）开发不同肠道微生物物种的阵列库，每个库都旨在报告存在独特的存在 目标聚糖，2。）开发快速的聚糖隔离管道，以净化感兴趣的单个基板 下游结构和功能表征，以及3.）开发具有转基因的微生物菌株 增强灵敏度或目标特异性。除了提供量化快速且廉价的替代方案 已知的聚糖，我们认为这些工具的进一步开发将揭示出大量的存在和丰富 以前无法检测到的聚糖并大大增强了我们对肠相互作用的理解 微生物及其哺乳动物宿主。