Deciphering Helicobacter pylori's glycocode: uncovering and harnessing drug targets

破译幽门螺杆菌的糖码：发现和利用药物靶点

• 批准号: 10351196
• 负责人: Danielle H. Dube
• 金额: $ 40.07万
• 依托单位: BOWDOIN COLLEGE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10351196
• 关键词: Address; Amino Sugars; Anabolism; Antibiotic Therapy; Antibiotics; Architecture; Area; Bacteria; Bacterial Antibiotic Resistance; Bacterial Infections; Biology; Cell surface; Cessation of life; Chemicals; Chemistry; Complex; Data; Defect; Deoxy Sugars; Development; Drug Targeting; Engineering; Enzymes; Gastritis; Gastrointestinal Diseases; Genes; Glycocalyx; Glycoproteins; Helicobacter Infections; Helicobacter pylori; Human; Immune; Immune Evasion; Infection; Knowledge; Label; Laboratories; Metabolic; Mission; Monosaccharides; Oligosaccharides; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Polysaccharides; Production; Protein Glycosylation; Public Health; Refractory; Reporter; Reporting; Research; Role; Stomach; Structure; System; Testing; Therapeutic Intervention; Translating; Ulcer; United States National Institutes of Health; Virulence Factors; Work; analog; base; cell envelope; emerging pathogen; fitness; glycoprotein biosynthesis; glycosylation; gut microbiome; gut microbiota; host colonization; human disease; inhibitor/antagonist; innovation; insight; microbial community; microbiota; mutant; new therapeutic target; pathogen; pathogenic bacteria; priority pathogen; sugar; tool; undergraduate student

Antibiotic resistant bacterial pathogens cause more than 2.8 million new infections and 32,000 deaths each year in the US,1 indicating the ineffectiveness of our existing arsenal of antibiotics. Even when antibiotics are effective at eradicating infection, most are inherently nonspecific for pathogens and have unintended conse- quences on beneficial microbiota.2 Bacterial cell surface glycans are compelling drug targets as they contain exclusively bacterial structures that are critical for strain fitness and pathogenesis.6-8 Despite the importance of glycan biosynthesis as a virulence factor, the systematic study and inhibition of bacterial glycans remains chal- lenging due to their utilization of rare deoxy amino sugars and their production of products that are refractory to traditional glycan analysis. The deployment of chemical tools to study bacterial glycans is a crucial step toward developing new glycosylation-based strategies to eradicate pathogenic infections. My laboratory seeks to apply metabolic oligosaccharide engineering (MOE) tools to diverse microbial communities and to develop new tools to study and perturb glycan biosynthesis in the gastric pathogen Helicobacter pylori (Hp). The central hypothe- sis of the application is that MOE with unnatural monosaccharides, including rare bacterial monosaccharide analogs11 and fluorescent inhibitors, will facilitate profiling glycopatterns in complex microbial communities, tracking of bacterial glycan biosynthesis, and disarming of bacteria by perturbing their glycan armor. Our hy- pothesis has been formulated on the basis of strong preliminary data produced in my laboratory, including the application of MOE to install detectable reporters into bacterial glycans,9-11 thus permitting the discovery of a protein glycosylation system,12, 13 the identification of glycosylation genes,14 and the development of metabolic glycan inhibitors.15 The proposed work will reveal new targets of therapeutic intervention, resulting in innovative approaches to treat bacterial infection. The central hypothesis will be tested by pursuing three specific aims. In Aim 1, metabolic glycan reporters will be used to profile and perturb glycopatterns in complex microbial com- munities, revealing information about glycan structure, underlying glycosylation machinery, and the selectivity with which bacteria can be targeted based on their distinctive glycan coating. In Aim 2, fluorescent substrate- based monosaccharide analogs will be used to track glycan biosynthesis intermediates and end products in Hp glycosylation mutants versus wildtype strains, yielding insight into how Hp glycosylation enzymes act together to produce glycans. In Aim 3, metabolic glycan inhibitors will disrupt glycoprotein biosynthesis in Hp, allowing us to sensitize Hp to host immune attack and potentiate the use of existing antibiotics. The proposed work will uncover bacterial glycans from the human gut microbiome that are amenable to study and inhibition with un- natural sugars, yield a bacterial glycomics approach to study glycan construction, and probe the effect of al- tered glycan architecture on immune recognition and antibiotic efficacy. Broadly, this work will introduce ap- proaches to study and harness glycans of priority pathogens to create urgently needed antibiotics.

抗生素耐药性细菌病原体导致280多万新感染和32，000人死亡 这表明我们现有的抗生素库无效。即使抗生素 有效根除感染，大多数是固有的非特异性病原体，并有意想不到的后果， 2细菌细胞表面聚糖是引人注目的药物靶标，因为它们含有 细菌结构对菌株适应性和发病机制至关重要。6 -8 聚糖生物合成作为一种毒力因子，细菌聚糖的系统研究和抑制仍然是挑战， 由于它们利用稀有的脱氧氨基糖和它们生产的产品是难处理的， 传统的聚糖分析。使用化学工具来研究细菌聚糖是实现这一目标的关键一步。 开发新的基于糖基化的策略以根除病原性感染。我的实验室想申请 代谢寡糖工程（莫伊）工具，使微生物群落多样化，并开发新的工具 目的研究和干扰幽门螺杆菌（Helicobacter pylori，Hp）多糖的生物合成。中央皮下组织- 该应用姊妹是具有非天然单糖的莫伊，包括稀有细菌单糖 类似物11和荧光抑制剂，将有助于在复杂的微生物群落中分析糖蛋白酶， 跟踪细菌聚糖的生物合成，以及通过扰乱细菌的聚糖装甲来解除细菌的武装。我们的hy- 假设已经制定了强有力的初步数据在我的实验室的基础上，包括 应用莫伊将可检测的报告基因安装到细菌聚糖中，9-11从而允许发现一种 蛋白质糖基化系统，12，13糖基化基因的鉴定，14和代谢的发展 15拟议的工作将揭示治疗干预的新靶点，从而产生创新的治疗方法。 治疗细菌感染的方法。中心假设将通过追求三个具体目标来检验。在 目的1、代谢性糖链报告基因将用于复杂微生物群落中糖链蛋白的分析和干扰。 的结构，揭示了有关聚糖结构，潜在的糖基化机制和选择性的信息。 基于其独特的聚糖涂层，可以用其靶向细菌。在目标2中，荧光底物- 的单糖类似物将用于追踪Hp中聚糖生物合成的中间体和终产物 糖基化突变体与野生型菌株，深入了解Hp糖基化酶如何共同作用 以产生聚糖。在目标3中，代谢聚糖抑制剂将破坏Hp中的糖蛋白生物合成， 我们可以使Hp对宿主免疫攻击敏感，并加强现有抗生素的使用。拟议的工作将 揭示来自人类肠道微生物组的细菌聚糖，这些细菌聚糖适合研究和抑制， 天然糖，产生细菌糖组学方法来研究聚糖结构，并探测al- 对免疫识别和抗生素功效的影响。从广义上讲，这项工作将介绍ap- 研究和利用优先病原体的聚糖来制造急需的抗生素。