Mucin Glycans in the Regulation of Microbial Virulence

项目概要/摘要 这个项目的目标是破译调节微生物毒力的粘蛋白聚糖密码。一层厚厚的， 水化良好的粘液是上皮衬里（例如口腔、胃肠道、胃肠道或胃肠道）上的关键防御机制。 气管和肺与粘蛋白相关的聚糖的特殊分子多样性和复杂性 聚合物是粘液的凝胶形成构件，几十年来一直被认识到。然而，他们的潜力 调节宿主及其相关微生物之间相互作用的机制几乎没有被利用， 聚糖的单独生物活性难以分析。我们在过去的资助期内的结果 支持粘蛋白聚糖通过调节跨界毒力在宿主保护中的核心作用;我们的结果 还强烈支持拟议的确定聚糖结构的努力的相关性和可行性， 负责抗毒性作用的机制。我们建议将联合收割机功能分析与自底向上相结合 粘蛋白样聚糖和聚合物的工程化，以揭示聚糖信号的设计原理， 粘蛋白调控编码。这些知识将使我们能够开始阐明并最终利用 聚糖和粘蛋白对微生物及其宿主的无数生物学后果。在目标1中，我们将收获 从分离的粘蛋白0-聚糖产生注释的文库，用于功能性的 从身体的主要粘膜表面进行分析，包括口腔、肺和消化道。这些 库将允许，第一次，功能研究，以获得深入了解的机制和化学O- 影响宿主-微生物相互作用的聚糖，如聚糖大小，特定残基序列，聚糖键， 和几何学在目标2中，我们将确定粘蛋白O-聚糖减弱两种病毒毒力的机制。 重要的人类粘膜病原体，铜绿假单胞菌和白色念珠菌，正成为 对治疗越来越有抵抗力。在目标3中，我们将描述粘蛋白O-聚糖的抗毒性作用 在完善的临床前体内感染模型中。在目标4中，我们将整合目标1中的知识- 3以先前无法达到的精确度设计优先的O-连接的糖基化粘蛋白样聚合物。这 该项目直接解决了一个紧迫的卫生保健问题：抗菌素耐药性正在迅速蔓延， 需要新的方法来对抗有问题的病原体。我们希望提供新的混凝土化学品 设计参数和分子来管理两种问题病原体， 抵抗治疗。多学科团队拥有将基础知识 临床前验证和尖端工程应用的科学问题：生物学家， 在粘液水凝胶系统的实验和理论专业知识，一个微生物学家与体内的专业知识， 感染模型，以及在受控聚糖和聚合物合成和表征方面具有专业知识的化学家。

PROJECT SUMMARY/ABSTRACT The goal of this project is to decipher the mucin glycan code that regulates microbial virulence. A layer of thick, well-hydrated mucus is a key defense mechanism on epithelial linings such as on the mouth, gastrointestinal tract, and lungs. The exceptional molecular diversity and complexity of glycans associated with mucin polymers, the gel-forming building blocks of mucus, has been recognized for decades. However, their potential for regulating interactions between a host and its associated microbes has barely been tapped because the individual bioactivities of glycans have been intractable to analysis. Our results from the past funding period support a central role for mucin glycans in host protection by regulating cross-kingdom virulence; our results also strongly support the relevance and feasibility of the proposed efforts to identify the glycan structures and mechanisms responsible for antivirulence effects. We propose to combine functional analysis with bottom-up engineering of mucin-like glycans and polymers to unravel the design principles of glycan signals and the mucin regulatory code. This knowledge will empower us to begin to elucidate, and ultimately harness, the myriad biological consequences of glycans and mucins on microbes and their hosts. In Aim 1, we will harvest bioactive glycans from mucins to generate annotated libraries from isolated mucin O-glycans for functional analysis from the major mucosal surfaces in the body, including the mouth, lungs, and digestive tract. These libraries will allow, for the first time, functional studies to obtain insight into mechanisms and chemistries of O- glycans that affect host-microbe interactions, such as glycan size, specific residue sequence, glycan linkages, and geometry. In Aim 2, we will identify the mechanisms by which mucin O-glycans attenuate virulence in two important human mucosal pathogens, Pseudomonas aeruginosa and Candida albicans, which are becoming increasingly resistant to treatment. In Aim 3, we will characterize the anti-virulence effects of mucin O-glycans in a well-established pre-clinical in vivo infection model. In Aim 4, we will integrate the knowledge from Aims 1- 3 to engineer prioritized O-linked glycosylated mucin-like polymers with previously unattainable precision. This project directly addresses an urgent health care problem: antimicrobial resistance is spreading rapidly and demands new approaches to combat problematic pathogens. We expect to deliver new concrete chemical design parameters and molecules to manage two problematic pathogens that are becoming increasingly resistant to treatments. The multidisciplinary team has the expertise necessary for combining fundamental science questions with pre-clinical validation and cutting-edge engineering applications: a biologist with experimental and theoretical expertise in mucus hydrogel systems, a microbiologist with expertise in in vivo infection models, and a chemist with expertise in controlled glycan and polymer synthesis and characterization.