Chemical Tools for Studying Fucosylated Glycans

用于研究岩藻糖基化聚糖的化学工具

基本信息

批准号：
9320789
负责人：
Peng Wu
金额：
$ 44.75万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9320789
关键词：
Adoptive Cell Transfers Apoptosis Biological Process Biology Breast Cancer Cell Breast Cancer cell line CD209 gene Cancer Immunology Science Cell Adhesion Cell physiology Cell surface Cells Chemicals Chemistry Chronic Lymphocytic Leukemia Clinic Collaborations Dendritic Cells Development Diphosphates Dorsal E-Selectin Embryo Embryonic Development Engineering Epithelial Epitopes Fertilization Fucose Fucosyltransferase Funding Grant Guanidines Guanosine Diphosphate Fucose Hematologic Neoplasms Human Image Immune Immune Cell Activation Immune System Diseases Immune system Immunosuppressive Agents In Situ In Vitro Inflammation Knowledge Leukocytes Ligands Link Lung Adenocarcinoma Lymphoma Malignant Neoplasms Malignant neoplasm of lung Mediating Medicine Membrane Proteins Mesenchymal Metabolic Methods Mus N-acetyllactosamine Neoplasm Metastasis Organism Pathologic Processes Patients Pattern Peripheral Physiological Processes Polysaccharides Process Production Proteins Recruitment Activity Regulation Role Sampling Signal Transduction Structure-Activity Relationship System T-Lymphocyte Tumor Immunity Up-Regulation WNT Signaling Pathway Work Zebrafish angiogenesis base college design early detection biomarkers fucose-binding protein glycosylation human disease in vivo leukocyte activation macrophage model development neoplastic cell public health relevance tool tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): Fucosylated glycans are involved in a variety of physiological and pathological processes in eukaryotic organisms including angiogenesis, fertilization, embryogenesis, cell adhesion, inflammation, and tumor metastasis. Some major gaps in our knowledge of fucosylated glycans include how they are dynamically regulated during embryo development and cancer progression, and their specific cellular functions. The broad objective of this project is to develop chemical tools to advance our understanding of the biological functions of fucosylated glycans in these processes. In the last granting period, we designed chemoenzymatic methods to synthesize guanidine 5′- diphosphate-β-L-fucose, the universal fucosyl donor, and chemically defined fucosides, as well as their structurally related derivatives. Using these chemical tools, we developed a few platforms to study the biological functions of fucosylated glycans both in vitro and in vivo, including a microarray platform to perform structure-activity relationship analysis of fucosides with fucose-binding proteins. In collaboration with Prof. Florence Marlow (Einstein), we developed a method to detect and image newly synthesized fucosides in zebrafish embryos using GDP-fucose functionalized with a bioorthogonal tag as the metabolic precursor. We discovered that up-regulation of N-linked fucosylation disrupts zebrafish dorsal-ventral patterning in the early embryogenesis by inhibiting the Wnt signaling pathway. Using a combination of fucosyltransferase and click chemistry, we developed a method to analysis the dynamic changes of cell-surface peripheral N- acetyllactosamine-bearing glycans in murine and human patient samples. These results validated the functional relevance of our chemoenzymatic tools for unraveling the biology of fucosylated glycans. Building upon this work, in the next granting period, we will invent new chemical tools, which will be used in conjunction with our established tools, to study the role of fucosylated glycans in the interaction between the immune system and tumors. In Aim 1, we will study the role of fucosylated glycans from the aspect of tumor cells. Specifically, we will investigate mechanisms by which tumor cells use fucosylated glycans to suppress immune cell activation (Aim 1). In the second part of this project, we will study the role of fucosylated glycas from the aspect of the immune system. We will comprehensively characterize the changes in cell-surface fucosylation during immune cell activation (Aim 2). Finally, we will design a method to engineer fucosylated glycans in immune cells and evaluate their anti-tumor activities (Aim 3). The chemical biology tools developed in this project will be made accessible to non-specialists for studying the biological functions of fucosides or the related glycans in their own systems.

描述（由申请人提供）：岩藻糖基化聚糖参与真核生物的多种生理和病理过程，包括血管生成、受精、胚胎发生、细胞粘附、炎症和肿瘤转移。我们对岩藻糖基化聚糖的认识中的一些主要空白包括它们如何发挥作用。在胚胎发育和癌症进展过程中受到动态调节，该项目的总体目标是开发化学工具来增进我们对细胞生物学功能的理解。在最后的授权期间，我们设计了化学酶方法来合成通用岩藻糖基供体 5'-二磷酸胍-β-L-岩藻糖和化学定义的岩藻糖苷，以及使用这些衍生物。化学工具，我们开发了一些平台来研究岩藻糖基化聚糖的体外和体内生物学功能，包括一个微阵列平台来进行结构-活性关系分析我们与 Florence Marlow 教授（爱因斯坦）合作，开发了一种使用生物正交标签功能化的 GDP-岩藻糖作为代谢前体来检测和成像斑马鱼胚胎中新合成的岩藻糖苷的方法。 -N-连锁岩藻糖基化的调节通过抑制 Wnt 信号通路来破坏早期胚胎发生中的斑马鱼背腹模式。利用岩藻糖基转移酶和点击化学，我们开发了一种方法来分析小鼠和人类患者样本中细胞表面外周 N-乙酰乳糖胺聚糖的动态变化。这些结果验证了我们的化学酶工具在揭示岩藻糖基化聚糖生物学方面的功能相关性。在这项工作的基础上，在下一个资助期内，我们将发明新的化学工具，这些工具将与我们现有的工具结合使用，以研究岩藻糖基化聚糖的作用在目标1中，我们将从肿瘤细胞的角度研究岩藻糖基化聚糖的作用，具体来说，我们将研究肿瘤细胞利用岩藻糖基化聚糖抑制免疫细胞活化的机制（目标1）。在该项目的第二部分中，我们将从免疫系统的角度研究岩藻糖基化的作用，我们将全面表征免疫细胞激活过程中细胞表面岩藻糖基化的变化。最后，我们将设计一种在免疫细胞中改造岩藻糖基化聚糖并评估其抗肿瘤活性的方法（目标3），该项目开发的化学生物学工具将可供非专业人士使用，以研究岩藻糖苷或岩藻糖苷的生物学功能。自己系统中的相关聚糖。