Development of Small Molecule Probes for the Selective Modification and Labeling of the Mycobacterial Cell Wall

开发用于选择性修饰和标记分枝杆菌细胞壁的小分子探针

• 批准号: 10394129
• 负责人: Daria Eldarovna Kim
• 金额: $ 1.93万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-06 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10394129
• 关键词: Academia; Address; Anabolism; Antibiotics; Antitubercular Antibiotics; Arabinose; Azides; Bacteria; Benign; Binding; Bypass; Carbohydrates; Cell Culture Techniques; Cell Wall; Cell surface; Cells; Cellular Assay; Cessation of life; Chemicals; Cord Factors; Corynebacterium; Coupling; Destinations; Development; Disease; Educational process of instructing; Environment; Enzymes; Fatty Acids; Flow Cytometry; Genus Mycobacterium; Glean; Glycolipids; Goals; Grant; Granuloma; Image; Immune; Immune response; Immune system; Infection; Infectious Agent; Institutes; Isotope Labeling; Label; Laboratories; Lactones; Learning; Leprosy; Life; Logic; Mannose; Mass Spectrum Analysis; Massachusetts; Mentors; Metabolic; Methods; Microbiology; Microscopy; Modification; Multi-Drug Resistance; Mycobacterium bovis; Mycobacterium leprae; Mycobacterium tuberculosis; Mycolic Acid; Organic Chemistry; Organism; Pathway interactions; Pharmaceutical Preparations; Play; Polysaccharides; Process; Proteomics; Protocols documentation; Reporting; Research; Resources; Signal Transduction; Specificity; Structure; Surface; System; Techniques; Technology; Testing; Training; Tuberculosis; Tuberculosis Vaccines; Virulence Factors; Visualization; Work; arabinofuranose; base; career; cell envelope; design; extracellular; glycosyltransferase; graduate student; human pathogen; immunoregulation; insight; lipoarabinomannan; live cell imaging; metabolic engineering; method development; monomer; mycobacterial; mycolate; novel strategies; pathogen; reconstitution; resistant strain; skills; small molecule; sugar; tool; treatment strategy; undergraduate student

PROJECT SUMMARY/ABSTRACT All cells throughout the three domains of life are coated in an array of structurally and compositionally diverse glycans which play a key role in cell signaling and recognition. In eukaryotic systems, a number of strategies for the perturbation of these structures with unnatural sugar probes have been disclosed. These probes have proven to be a powerful tool for interrogating glycan structure, function and dynamics in living systems. However, because these techniques all rely on extensive metabolic processing of the probe prior to incorporation, they have been largely ineffective in bacteria, which utilize a multitude of catabolic pathways that can degrade and reconstitute parts of the probe on the way to the final destination. This proposal aims to circumvent this problem by designing small molecule probes that engage directly with the extracellular enzymes that construct important cell wall motifs. This strategy, termed ‘biosynthetic incorporation’ will be applied to the selective modification of two important glycolipid virulence factors within the cell wall of Mycobacterium tuberculosis (Mtb), the lipoarabinomannan (LAM) and trehalose dimycolate (TDM), which help the pathogen infect host cells and evade the body’s immune defense system. This proposal focuses primarily on the design and synthesis of azide-modified arabinofuranose, mannose and mycolic acid probes and studies intended to quantify the degree of incorporation and distribution of the probes. The tools developed in the course of this work will be used to visualize the localization and dynamics of the LAM and TDM during host cell infection using live cell imaging. Insights gleaned from these studies could elucidate new strategies for the treatment of mycobacterial diseases like Mtb. The Kiessling research group at the Massachusetts Institute of Technology (MIT) is an ideal environment to pursue the research outlined in this proposal and to achieve the training goals that will prepare me for a career in academia. In the Kiessling group, I will be able to continue to hone and utilize my skills in synthetic organic chemistry while also gaining expertise in a diverse array of new techniques including microscopy, microbiology, flow cytometry and cell culture. I will have the opportunity to mentor undergraduates and graduate students within the group. In addition to this, the MIT Teaching and Learning Lab provides programming and a number of resources that I will utilize in order to continue to build my teaching skills. I am confident that the exceptional laboratory and institutional environment at MIT will provide me with the tools necessary for a successful independant career.

项目概要/摘要 生命三个领域的所有细胞都被一系列结构和成分所覆盖 多种聚糖在细胞信号传导和识别中发挥关键作用。在真核系统中，许多 已经公开了用非天然糖探针扰动这些结构的策略。这些 探针已被证明是研究生命体中聚糖结构、功能和动态的强大工具 系统。然而，因为这些技术都依赖于探针在使用之前的广泛代谢处理。 掺入后，它们对细菌基本上无效，细菌利用多种分解代谢途径 可以在到达最终目的地的途中降解和重建探针的某些部分。该提案旨在 通过设计直接与细胞外酶结合的小分子探针来解决这个问题 构建重要的细胞壁基序。这一战略被称为“生物合成整合”，将应用于 选择性修饰分枝杆菌细胞壁内两种重要的糖脂毒力因子 结核病 (Mtb)、阿拉伯脂甘露聚糖 (LAM) 和海藻糖二霉菌酸酯 (TDM)，有助于病原体 感染宿主细胞并逃避人体的免疫防御系统。本提案主要关注设计 叠氮化物修饰的阿拉伯呋喃糖、甘露糖和霉菌酸探针的合成和研究旨在 量化探针的掺入和分布程度。在这项工作过程中开发的工具 将用于使用活体观察宿主细胞感染期间 LAM 和 TDM 的定位和动态 细胞成像。从这些研究中收集的见解可以阐明治疗的新策略 分枝杆菌疾病，如 Mtb。 麻省理工学院 (MIT) 的 Kiessling 研究小组是一个理想的环境 进行本提案中概述的研究并实现为我的职业生涯做好准备的培训目标 在学术界。在基斯林集团，我将能够继续磨练和利用我在合成有机方面的技能 化学，同时还获得了各种新技术的专业知识，包括显微镜、微生物学、 流式细胞术和细胞培养。我将有机会指导本科生和研究生 组内。除此之外，麻省理工学院教学实验室还提供编程和许多 我将利用这些资源来继续提高我的教学技能。我坚信，非凡的 麻省理工学院的实验室和机构环境将为我提供成功所需的工具 独立的职业生涯。