Unraveling the enzymatic pathway of gut bacterial mucus degradation to treat inflammation

揭示肠道细菌粘液降解的酶促途径以治疗炎症

• 批准号: 10681374
• 负责人: Nicole M Koropatkin
• 金额: $ 55.4万
• 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: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10681374
• 关键词: Animals; Anti-Inflammatory Agents; Antigens; Bacteria; Bacterial Physiology; Bacteroides; Bacteroides thetaiotaomicron; Biological Assay; Biology; Calorimetry; Carbohydrates; Cell surface; Childhood; Chronic Disease; Citrobacter rodentium; Colitis; Colon; Colorectal Cancer; Communities; Complex; Complex Mixtures; Crystallography; Data; Development; Diet; Dietary Fiber; Discipline; Disease; Early identification; Ecosystem; Enzymatic Biochemistry; Enzyme Inhibition; Enzymes; Epithelium; Event; Family; Fiber; Future; Gene Expression Profiling; Genetic Polymorphism; Germ-Free; Glycobiology; Glycoproteins; Glycoside Hydrolases; Gnotobiotic; Goals; Goblet Cells; Growth; Health; Human; In Vitro; Individual; Infection; Inflammation; Inflammatory Bowel Diseases; Interleukin-10; Intestines; Knowledge; Link; Measures; Microbial Physiology; Modeling; Molecular; Molecular Biology; Molecular Weight; Monosaccharides; Mucins; Mucous Membrane; Mucous body substance; Mus; Mutation; Nutrient; Oligonucleotides; Pathway interactions; Peptide Hydrolases; Play; Polysaccharides; Predisposition; Process; Proteins; Recombinants; Research; Research Personnel; Resort; Role; Series; Signal Transduction; Source; Specificity; Structure; Substrate Specificity; Sulfatases; Sulfate; Surface; System; Testing; Therapeutic; Thick; Tissues; Titrations; Vertebral column; Wild Type Mouse; Work; combinatorial; commensal microbes; cytokine; design; enzyme activity; experience; experimental study; fitness; gastrointestinal; glycosylation; gut bacteria; gut microbes; gut microbiota; host-microbe interactions; human disease; improved; in vivo; in vivo Model; inhibitor; interest; loss of function; member; microbial; microbial community; microbiota; mutant; novel; pathogen; prevent; structural biology; synergism; tool; transcriptome sequencing; whole genome

Summary The composition and physiology of the microbial community (microbiota) in the human colon has been linked to a number of diseases. Mechanistic details for most of these interactions are still badly needed. The shared focus of the four investigators assembled to conduct the proposed project is to understand how gut microbes interact with and metabolize complex carbohydrates—especially the glycans attached to secreted host mucus. Mucus is the first barrier that separates intestinal bacteria from host tissue and is a complex mixture of secreted mucin glycoprotein and other molecules. Some bacteria have evolved to forage on mucus as a source of nutrients. We have previously shown that this mucus foraging activity increases when exogenous dietary fiber polysaccharides are absent. Using a gnotobiotic model of fully sequenced human gut bacteria, we have shown that during fiber deficiency the gut microbiota resorts to degrading mucus for nutrients, leading to erosion of its integrity. In wild-type mice, a reduced mucus barrier increases epithelial access and lethal colitis by the mucosal pathogen, Citrobacter rodentium. More strikingly, when this same synthetic microbiota is assembled in mice deficient in interleukin 10, a cytokine for which loss of function is associated with human pediatric inflammatory bowel disease (IBD), animals develop lethal inflammation in the absence of pathogen, but only on a low fiber diet. Our work has therefore revealed functional connections between mucus integrity, diet and gut microbes in precipitating IBD. The complete deconstruction of mucin glycoproteins requires a consortium of enzymes: peptidases to hydrolyze the protein backbone and sulfatases and glycoside hydrolases that recognize sulfated or unsulfated oligo- and monosaccharides within discrete glycosidic linkage contexts. Our central hypothesis is that mucin is degraded in a series of sequential steps by individual activities in this enzyme consortium and that essential catalytic steps exist, which may be contributed by different species that work synergistically to degrade mucus. We will test this hypothesis by first defining the sequential action, positional specificity and key structural facets of bacterial enzymes required for degradation of gastrointestinal mucins. We will use sequential and combinatorial treatments of various forms of mucin with pure recombinant enzymes, which we have already identified in the members of our synthetic microbiota. In parallel, we will measure the requirement for individual, discrete mucus-degrading steps within genetically- manipulable model species using in vitro and mouse in vivo models as readouts. The research team is composed of four leaders in the disciplines of gut bacterial physiology and molecular biology, structural biology and enzymology, mucin biology and glycoanalytics, all with a shared interest in the mechanisms of mucus degradation and the consequences for human disease. Successful completion of these experiments will define a precise series of mechanistic steps for bacterial mucin degradation and could lead to therapies to limit these events in diseases like IBD.

概括 人类结肠中微生物群落（微生物群）的组成和生理学已连接 对于大多数互动的机械细节仍然是不太需要的 四个调查人员组装以进行支撑项目的重点是了解肠道微生物如何 与分泌的宿主粘液相互作用并代谢，尤其是与分泌的宿主粘液相互作用。 粘液是将肠道细菌与宿主组织分离的第一个障碍物，并且是复杂的混合物 分泌的粘蛋白糖蛋白和其他分子。 营养的来源。 使用完全测序的人类肠道细菌的饮食中没有纤维化糖。 已经表明，在纤维缺乏期间，肠道微生物群诉诸降解营养的粘液，也导致 野生型小鼠的侵蚀，粘液屏障的上皮通道降低 通过粘膜病原体，柠檬酸杆菌更加惊人。 在白介素10中的小鼠中汇集，抗人功能丧失的抗体因子 小儿炎性肠病（IBD），动物在没有病原体的情况下会出现致命的炎症 但是，只有低纤维饮食。 饮食和肠道微生物沉淀IBD。 酶的财团：肽群，将蛋白质主链和磺胺糖和糖苷和糖化 在离散糖苷键内识别硫化或未剥离的寡糖和单糖的水解酶 我们的核心假设是粘蛋白通过单个活动进行了一系列依次 在这个酶联盟中，存在累积的催化步骤，这可能由不同的贡献 与降解粘液降解的协同主义的物种我们将首先抗衡顺序。 降解所需的细菌酶的活动特异性和关键结构方面 胃肠道粘蛋白。 纯重组酶，我们在合成微生物群的成员中鉴定出了小巷 平行于，我们将衡量对基因的单个，离散粘液降解步骤的要求 使用体外和小鼠在体内模型的可操纵模型作为读数。 由迪斯科细菌生理学和分子生物学的四个领导者组成，结构生物学 以及酶学，粘蛋白生物学和甘油分析，都对粘液机制具有共同的兴趣 降解和人类疾病的后果将定义 一个精确的机械步骤，用于粘粘粘膜降解，可能会限制这些 IBD等疾病的事件。

项目成果

Unravelling specific diet and gut microbial contributions to inflammatory bowel disease.

揭示特定饮食和肠道微生物对炎症性肠病的影响。

• DOI: 10.21203/rs.3.rs-2518251/v1
• 发表时间: 2023
• 期刊: Research square
• 作者: Pereira,GabrielVasconcelos;Boudaud,Marie;Wolter,Mathis;Alexander,Celeste;DeSciscio,Alessandro;Grant,EricaT;Trindade,BrunoCaetano;Pudlo,NicholasA;Singh,Shaleni;Campbell,Austin;Shan,Mengrou;Zhang,Li;Willieme,Stéphanie;Kim,Kwi
• 通讯作者: Kim,Kwi

Deprivation of dietary fiber in specific-pathogen-free mice promotes susceptibility to the intestinal mucosal pathogen Citrobacter rodentium.

• DOI: 10.1080/19490976.2021.1966263
• 发表时间: 2021-01
• 期刊: Gut microbes
• 影响因子: 12.2
• 作者: Neumann M;Steimle A;Grant ET;Wolter M;Parrish A;Willieme S;Brenner D;Martens EC;Desai MS
• 通讯作者: Desai MS

Diverse events have transferred genes for edible seaweed digestion from marine to human gut bacteria.

• DOI: 10.1016/j.chom.2022.02.001
• 发表时间: 2022-03-09
• 期刊: CELL HOST & MICROBE
• 影响因子: 30.3
• 作者: Pudlo, Nicholas A.;Pereira, Gabriel Vasconcelos;Parnami, Jaagni;Cid, Melissa;Markert, Stephanie;Tingley, Jeffrey P.;Unfried, Frank;Ali, Ahmed;Varghese, Neha J.;Kim, Kwi S.;Campbell, Austin;Urs, Karthik;Xiao, Yao;Adams, Ryan;Martin, Duna;Bolam, David N.;Becher, Dorte;Eloe-Fadrosh, Emiley A.;Schmidt, Thomas M.;Abbott, D. Wade;Schweder, Thomas;Hehemann, Jan Hendrik;Martens, Eric C.
• 通讯作者: Martens, Eric C.

Sulfated glycan recognition by carbohydrate sulfatases of the human gut microbiota.

• DOI: 10.1038/s41589-022-01039-x
• 发表时间: 2022-08
• 期刊: Nature chemical biology
• 影响因子: 14.8
• 作者: Luis AS;Baslé A;Byrne DP;Wright GSA;London JA;Jin C;Karlsson NG;Hansson GC;Eyers PA;Czjzek M;Barbeyron T;Yates EA;Martens EC;Cartmell A
• 通讯作者: Cartmell A