Chemoenzymatic glycan editing for deciphering biological functions of glycans

化学酶聚糖编辑破译聚糖的生物学功能

基本信息

批准号：
10329937
负责人：
Peng Wu
金额：
$ 76.42万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10329937
关键词：
Affinity Autoimmune Diseases Binding Binding Proteins Biological Process Cell Communication Cell physiology Cell surface Cells Development Disease Progression Engineering Environment FDA approved Fucose Genome Glycoconjugates Glycoproteins Goals Immune Immune System Diseases Immune checkpoint inhibitor Immune signaling Immunoglobulins Immunotherapy In Situ Label Lectin Ligands Malignant Neoplasms Mediating Metabolic Methods Modality Modification Molecular Molecular Biology Techniques Monosaccharides N-acetyllactosamine Natural Killer Cells Oligosaccharides Organism Patients Polysaccharides Production Property Recombinants Role Sialic Acids Signal Transduction Specificity Structure T-Lymphocyte Therapeutic Agents Tumor Immunity Vaccination Visualization anti-CTLA4 anti-PD-1 base checkpoint inhibition cytokine cytotoxic exhaustion glycosylation glycosyltransferase human disease immune activation immunoregulation interest novel novel therapeutics receptor receptor binding response sialic acid binding Ig-like lectin sugar nucleotide tool

项目摘要

PROJECT SUMMARY/ABSTRACT Cell-surface glycans participate in numerous biological processes, including signal transduction, cell-cell communication and development. Aberrant glycosylation is a hallmark of human disease. At a molecular level, glycans represent the first points of contact between cells. However, not directly encoded in the genome, these biomolecules are challenging to study using molecular biology techniques alone. Metabolic oligosaccharide engineering (MOE) developed in late 1990’s has revolutionized the way for the labeling and visualization of glycans in living organisms. In this method, cells’ own glycan biosynthetic machinery is hijacked to incorporate unnatural monosaccharides with linkage promiscuity. Complementary to MOE, chemoenzymatic glycan editing has emerged as a valuable tool to probe and modify glycan structures within a cellular environment. Unlike MOE, chemoenzymatic glycan modification utilizes recombinant glycosyltransferases to transfer natural or unnatural monosaccharides with novel functions from activated nucleotide sugars to glycoconjugates on the cell surface with linkage specificity. For these reasons, chemoenzymatic glycan modification provides a facile and more precise way for probing the function of glycans in their native environments. Building upon our successful application of chemoenzymatic glycan editing, in the next five years we will expand our chemoenzymatic tool kits to study glycans’ cellular functions with a focus on the special roles of N- acetyllactosamine (LacNAc), fucose and sialic acid in immune regulation. Cell-surface LacNAc mediates ligand-receptor binding and sets a threshold for initiating the downstream signaling for immune cell activation. LacNAc residues are dynamically modified by sialic acid and/or fucose. However, the specific roles of these modifications in immune regulation and disease progression remain obscure. We are particularly interested in finding out: (1) if changes in LacNAc and fucosylation status can serve as glycan signatures of T cell exhaustion during which T cells gradually lose their cytokine production, proliferation and cytotoxic capacity; (2) Can cell-surface in situ LacNAc fucosylation be used to boost the efficacy of antitumor immunity of T cells and NK cells? In parallel, we will develop chemoenzymatic tools for profiling sialylated glycoprotein ligands of Siglecs (sialic acid-binding immunoglobulin-type lectins) and for the identification of unnatural, high-affinity and specific ligands to interrogate Siglec functions. Through these studies, we will gain a deeper understanding of how LacNAc, fucose and sialic acid are involved in the regulation of the immune cell activation, effector function and exhaustion. Tools developed in this project can also be used to study other types of glycans and their interactions with glycan binding proteins.

项目摘要/摘要细胞表面聚糖参与许多生物学过程，包括信号转导，细胞细胞沟通和发展。异常糖基化是人类疾病的标志。在分子水平上，聚糖代表细胞之间接触的第一点。但是，不是直接编码的基因组，而是仅使用分子生物学技术来研究生物分子。代谢寡糖工程（MOE）于1990年代末开发，彻底改变了标签和可视化的方式生物中的聚糖。在这种方法中，细胞自己的聚糖生物合成机械被劫持以合并不自然的单糖和连锁杂交。互补的MOE，化学酶聚糖编辑已成为探测和在细胞环境中修改聚糖结构。与MOE不同，化学酶聚糖修饰利用重组糖基转移酶以新功能转移天然或不自然的单糖从活化的核丁基糖到具有连锁特异性在细胞表面上的糖缀合物。为此原因，化学酶聚糖的修饰为探测功能提供了一种简便，更精确的方法在其本地环境中的聚糖。在我们成功应用化学酶聚糖编辑的基础上，在接下来的五年中，我们将扩展我们的化学酶工具套件以研究聚糖的细胞功能，重点是N-的特殊作用免疫调节中的乙酸酯（LACNAC），岩藻糖和唾液酸。细胞表面LACNAC培养基配体受体结合并设置了启动下游信号以进行免疫电池激活的阈值。 LACNAC保留通过唾液酸和/或fucose动态修饰。但是，这些角色的特定角色免疫调节和疾病进展的修改仍然晦涩难懂。我们特别有兴趣找出：（1）如果LACNAC和岩藻糖基化状态的变化可以服务作为T细胞耗尽的聚糖特征，在此期间T细胞逐渐失去细胞因子的产生，增殖和细胞毒性能力；（2）可以使用细胞表面的原位lacnac fecosylation来提高 T细胞和NK细胞抗肿瘤免疫的功效？同时，我们将开发用于化学酶的工具分析siglecs（唾液酸结合免疫球蛋白型讲座）的溶解糖蛋白配体，用于鉴定不自然，高亲和力和特定配体以询问Siglec功能。通过这些研究，我们将对LACNAC，Fucose和Sialic Acid的更深入了解参与了免疫细胞激活，效应子功能和衰竭的调节。开发了工具该项目还可以用于研究其他类型的聚糖及其与聚糖结合蛋白的相互作用。