Peptidoglycan Fragment Library to Investigate Innate Immune Responses

用于研究先天免疫反应的肽聚糖片段库

• 批准号: 10034684
• 负责人: Catherine Leimkuhler Grimes
• 金额: $ 51.66万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-03 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10034684
• 关键词: Adjuvant; Affinity; Amino Acids; Amino Sugars; Antibiotic Therapy; Antibiotics; Bacteria; Binding; Biological; Biological Markers; Biology; Bone Marrow; Carbohydrates; Cell Wall; Cells; Chemicals; Communities; Complex; Data; Development; Disaccharides; Discipline; Disease; Elements; Enzymes; Event; Family member; Functional disorder; Gene Expression Profile; Generations; Glucosamine; Goals; Health; Healthcare; Host Defense; Human; Immune; Immune response; Immune signaling; Immune system; Immunologic Receptors; Immunologics; Immunology procedure; Immunotherapy; Inflammatory; Inflammatory Bowel Diseases; Innate Immune Response; Innate Immune System; Intracellular Membranes; Investigation; Label; Libraries; Ligands; Link; Maps; Methodology; Modification; Monosaccharides; Muramic Acid; Natural Immunity; Organism; Osmotic Pressure; Pathogenicity; Pathway interactions; Peptides; Peptidoglycan; Phosphotransferases; Polymers; Polysaccharides; Positioning Attribute; Printing; Production; Property; Receptor Signaling; Reproducibility; Research Personnel; Role; Signal Pathway; Signal Transduction; Specificity; Structure; Surface; Therapeutic; Vertebral column; adaptive immunity; base; chemical synthesis; crosslink; design; dimer; drug development; effective therapy; genome-wide; innate immune pathways; macrophage; member; microbial; microbiota; novel; pathogenic bacteria; peptidoglycan receptor; preference; receptor; receptor binding; response; screening; tool; trafficking; transcription factor; transcriptome; vector

PROJECT SUMMARY Bacterial cells surround themselves with a peptidoglycan (PG) cell wall, an essential structure that resists changes in osmotic pressure and other environmental insults. The goal of this proposal is to develop a PG fragment library that captures the natural diversity to facilitate the proper innate and adaptive immunity biological studies. PG fragments are used by the innate immune system to correctly recognize and respond to the presence of bacteria. We hypothesize that PG diversity naturally present across the multitude of bacterial species is essential for generating the correct immune response for host defense. Study of these important PG fragments has been hampered by the lack of reproducible, high purity compounds. Currently, researchers are limited to few carbohydrate probes, such as MDP, and even fewer larger fragments. We propose to functionalize three carbohydrate cores: monosaccharide, disaccharide and peptide dimers, with select amino acids which are highly represented across a variety of bacterial species. The synthetic workflow is modular, builds off our expertise in producing highly pure PG fragments and permits for modification with chemical biology probes at multiple pinpoints on the library members. A large PG fragment library of this type has not been produced before. To overcome this challenge we propose to use common intermediates for all three carbohydrate cores. We aim to use this library to interrogate innate immune responses elicited in macrophages. In Aim One, a large-scale, modular synthetic effort will be undertaken to produce the >400 member library. For Aim Two the disaccharide and peptide linked dimer family members will be employed in an unbiased genome wide transcriptome analysis to unravel gene expression signatures that define the responses to PG classes in bone marrow derived macrophages. We will synthesize the chemical biology probes to investigate receptors and signaling partners. Aim Three will probe the innate immune response of the PG through the production of a PG-microarray. This array will fix the PG derivatives in multiple orientations permitting the sweeping of potential receptors across chemical space. The arrays will be used to assess the substrate binding preferences of a variety of innate immune receptors indicated in PG recognition. This library will accurately capture the PG fragment diversity, providing powerful probes for the proposed immunological assays and new tools for the microbial and immunological communities.

项目摘要 细菌细胞用肽聚糖（PG）细胞壁包围，这是一种必需结构 渗透压和其他环境侮辱的变化。该提议的目的是开发PG 碎片库捕获自然多样性，以促进适当的先天和适应性免疫力 生物学研究。先天免疫系统使用PG片段来正确识别和回应 存在细菌。我们假设PG多样性自然存在于许多细菌中 物种对于为宿主防御产生正确的免疫反应至关重要。这些重要的研究 缺乏可重复的高纯度化合物，PG碎片受到了阻碍。目前，研究人员 仅限于很少的碳水化合物探针，例如MDP，甚至更少的较大碎片。我们建议 功能化三个碳水化合物核：单糖，二糖和肽二聚体，带有精选的氨基 在多种细菌种类中高度代表的酸。合成工作流是模块化的， 建立我们在生产高度纯PG片段和使用化学修饰的许可方面的专业知识。 在图书馆成员上的多个查明点上的生物学探针。这种类型的大型PG碎片库还没有 是之前生产的。为了克服这一挑战，我们建议使用这三个挑战 碳水化合物核心。我们的目的是利用该图书馆询问引起的先天免疫反应 巨噬细胞。在目标中，将采取大规模的模块化合成努力，以生产> 400 成员库。对于目标两个 公正的基因组广泛的转录组分析，以揭示定义的基因表达特征 对骨髓中PG类的反应。我们将合成化学生物学 研究受体和信号伴侣的探针。 AIM三将探究先天免疫反应 PG通过PG微阵列的生产。此数组将在多个方向上修复PG衍生物 允许跨化学空间扫除潜在受体。阵列将用于评估 PG识别中指示的各种先天免疫受体的底物结合偏好。这个库 将准确捕获PG片段多样性，为拟议的免疫学提供强大的探针 微生物和免疫社区的测定和新工具。