Chemical Tools for Studying Fucosylated Glycans

用于研究岩藻糖基化聚糖的化学工具

• 批准号: 9105947
• 负责人: Peng Wu
• 金额: $ 47.06万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 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/9105947
• 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; Hematologic Neoplasms; Human; Image; Immune; Immune Cell Activation; Immune System Diseases; Immune system; Immunity; 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; Modeling; Mus; N-acetyllactosamine; Neoplasm Metastasis; Organism; Pathologic Processes; Patients; Pattern; Peripheral; Physiological Processes; Polysaccharides; Process; Production; Proteins; Recruitment Activity; Regulation; Role; Sampling; Signal Pathway; Signal Transduction; Structure-Activity Relationship; System; T-Lymphocyte; Up-Regulation; Work; Zebrafish; angiogenesis; base; college; design; early detection biomarkers; fucose-binding protein; glycosylation; human disease; in vivo; leukocyte activation; macrophage; neoplastic cell; public health relevance; tool; tumor; tumor progression

 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(申请人提供)：岩藻糖化多聚糖在真核生物中参与多种生理和病理过程，包括血管生成、受精、胚胎发生、细胞黏附、炎症和肿瘤转移。在我们对岩藻糖化多糖的了解中，一些主要的空白包括它们在胚胎发育和癌症进展过程中是如何动态调节的，以及它们特定的细胞功能。这个项目的广泛目标是开发化学工具，以促进我们对岩藻糖化多糖在这些过程中的生物学功能的理解。在最后一个授权期，我们设计了化学酶方法合成了通用岩藻糖供体5‘-二磷酸-β-L-岩藻糖、岩藻糖苷及其结构相关的衍生物。利用这些化学工具，我们开发了几个平台来研究岩藻糖化多糖的体内外生物学功能，包括一个进行岩藻糖苷与岩藻糖结合蛋白构效关系分析的微阵列平台。与弗洛伦斯·马洛教授(爱因斯坦)合作，我们开发了一种检测和成像斑马鱼胚胎中新合成的岩藻糖苷的方法，该方法使用带有生物正交标签的GDP-岩藻糖作为代谢前体。我们发现，上调N-连接岩藻糖基化通过抑制Wnt信号通路，破坏了斑马鱼早期胚胎发育的背腹模式。利用岩藻糖基转移酶和点击化学相结合的方法，我们发展了一种分析小鼠和人类患者样本中细胞表面含N-乙酰乳糖胺的糖链的动态变化的方法。这些结果验证了我们的化学酶工具在解开岩藻糖化多糖生物学方面的功能相关性。在这项工作的基础上，在下一个授权期，我们将发明新的化学工具，这些工具将与我们现有的工具一起使用，以研究岩藻糖化多糖在免疫系统和肿瘤之间相互作用中的作用。在目标1中，我们将从肿瘤细胞的角度研究岩藻糖化多糖的作用。具体地说，我们将研究肿瘤细胞使用岩藻糖化多糖抑制免疫细胞激活的机制(目标1)。在本项目的第二部分，我们将从免疫系统的角度研究岩藻糖化糖的作用。我们将全面描述免疫细胞激活过程中细胞表面岩藻糖化的变化(目标2)。最后，我们将设计一种在免疫细胞中设计岩藻糖化多糖的方法，并评估它们的抗肿瘤活性(目标3)。在该项目中开发的化学生物学工具将向非专业人员提供，以研究岩藻糖苷或相关多糖在其自身系统中的生物功能。