Core Fucosylation of N-linked Glycans as a Regulator of CD8+ T cell Activation and Function

N 连接聚糖的核心岩藻糖基化作为 CD8 T 细胞激活和功能的调节剂

基本信息

批准号：
10333397
负责人：
Jeffrey C. Nolz
金额：
$ 19.25万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
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：
2021-01-25 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10333397
关键词：
Affinity Aleuria aurantia lectin Amino Acids Antigens Apoptosis Asparagine Binding Biochemical Biological Bone Marrow CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes Carbohydrates Cell Adhesion Cell physiology Cells Chimera organism Chronic Cytotoxic T-Lymphocytes DNA Data Development Enterobacteria phage P1 Cre recombinase Enzymes Fucose Fucosyltransferase Functional disorder Generations Genetic Code Goals Immunotherapy In Vitro Individual Infection Knockout Mice Label Lectin Ligands Link Lymphocytic choriomeningitis virus Mass Spectrum Analysis Mediator of activation protein Membrane Proteins Modification Molecular Mus Peptides Physiology Play Polysaccharides Post-Translational Protein Processing Production Proliferating Proteins RNA Reagent Role Signal Transduction Structure T cell response T-Cell Activation T-Lymphocyte Techniques Testing Tissues Translating Vaccine Design Virus Diseases anti-PD-L1 antigen-specific T cells base conditional knockout cytokine exhaust exhaustion functional restoration gene product glycosylation glycosyltransferase improved in vivo interleukin-21 receptor mammalian genome novel preservation protein degradation protein function receptor

项目摘要

PROJECT SUMMARY/ABSTRACT Post-translational carbohydrate modifications are important mediators of several cellular processes including protein turnover, cell adhesion, signal transduction, modulating receptor affinity for ligand, and apoptosis. However, in contrast to the well-established genetic code where biological information in DNA results in the generation of RNA which is then translated into protein, no such paradigm exists for predicting the vast array of possible post-translational glycosylation structures that could potentially decorate a given gene product. One such glycan modification has been defined as ‘core fucosylation’ of N-linked glycans, which occurs when L- fucose is covalently linked via an a1,6 linkage to the initial N-acetylglucosamine that originates from the asparagine amino acid. Although core fucosylation of N-linked glycans is necessary for normal development and physiology, whether these N-linked glycans are critical regulators of T cell activation and/or function is largely unknown. Here, we show that CD8+ T cells are decorated with N-linked glycans containing a core fucose and that the abundance of this specialized glycan modification increases significantly following their activation both in vitro and in vivo. Fucosyltransferase 8 (Fut8; a1,6-fucosyltransferase) is the only glycosyltransferase enzyme in the mammalian genome that can facilitate core fucosylation of N-linked glycans and we have now generated a novel Fut8 conditional knockout mouse that allows us to eliminate expression of Fut8 in a cell-specific manner. Using a T cell-specific cre-recombinase, our preliminary data show that expression of Fut8 and the subsequent generation of core fucosylated N-linked glycans is essential to maintain antigen-specific CD8+ T cell function (e.g. production of cytokines) during chronic viral infection. Here, we propose to use our new reagent to 1) identify the landscape of proteins expressed by activated CD8+ T cells that become decorated with core fucosylated N- linked glycans using a mass spectrometry approach and 2) to subsequently determine the biological relevance of this form of post-translational glycosylation in maintaining the function of antigen-specific CD8+ T cells during chronic viral infection.

项目摘要/摘要翻译后碳水化的修饰是几个细胞过程的重要介体蛋白质更新，细胞粘合剂，信号转导，调节配体的受体亲和力和凋亡。但是，与公认的遗传密码相反，DNA中的生物学信息导致然后将RNA转化为蛋白质，没有这种范式来预测大量可能会在可能装饰给定基因产物的转换后糖基化结构。一这种聚糖的修饰已被定义为N连接聚糖的“核心构造”，当L-发生时发生通过A1,6链接与初始N-乙酰葡萄糖的链接共价连接天冬酰胺氨基酸。尽管N连锁聚糖的核心脉络化化对于正常发育和生理学，这些N连接的聚糖是否是T细胞激活和/或功能的关键调节剂未知。在这里，我们表明CD8+ T细胞用含有核心fucose和的N连接的聚糖装饰这种专业的聚糖修饰的抽象在激活后都大大增加体外和体内。岩藻糖基转移酶8（FUT8; A1,6-羟基转移酶）是唯一的糖基转移酶在可以促进N连接聚糖的核心构造化的哺乳动物基因组中，我们现在已经产生一种新型的FUT8条件敲除小鼠，使我们能够以细胞特异性的方式消除FUT8的表达。使用T细胞特异性的CRE聚合酶，我们的初步数据表明FUT8的表达和随后的表达核心基化的N连接聚糖的产生对于维持抗原特异性CD8+ T细胞功能至关重要（例如，在慢性病毒感染过程中产生细胞因子）。在这里，我们建议将新试剂用于1）确定活化的CD8+ T细胞表达的蛋白质的景观，这些细胞被核心基化的N-装饰使用质谱法连接的聚糖和2）随后确定生物学相关性这种形式的翻译后糖基化在维持抗原特异性CD8+ T细胞功能方面的功能慢性病毒感染。