T cell regulation by N-glycosylation

通过 N-糖基化调节 T 细胞

• 批准号: 8650769
• 负责人: MICHAEL DEMETRIOU
• 金额: $ 36.78万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-12-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8650769
• 关键词: 129/Sv Mouse; Affinity; Alleles; Amino Acid Sequence; Anabolism; Animal Model; Antigens; Autoimmune Diseases; Autoimmunity; Binding; Carbohydrates; Cell surface; Child; Cholesterol; Complex; DNA; Data; Disease; Endocytosis; Environment; Enzymes; Experimental Autoimmune Encephalomyelitis; F-Actin; Galactose Binding Lectin; Galectin 3; Gene Expression; Genetic; Glycoproteins; Golgi Apparatus; Growth; Haplotypes; Hexosamines; Human; Human Genetics; Hyperactive behavior; Immune; Inbred NOD Mice; Inbred Strains Mice; Individual; Inflammatory Bowel Diseases; Kidney; LCP2 gene; Life; Ligands; Macromolecular Complexes; Medial Golgi; Mediating; Membrane; Membrane Glycoproteins; Metabolic; Metabolic Control; Molecular; Molecular Structure; Multiple Sclerosis; Mus; N-Acetylglucosaminyltransferases; PL/J Mouse; PTPRC gene; Pathway interactions; Peptide Sequence Determination; Phenotype; Polysaccharides; Predisposition; Protein Glycosylation; Protein Tyrosine Phosphatase; Proteins; Receptor Signaling; Regulation; Regulatory T-Lymphocyte; Resistance; Rheumatoid Arthritis; Risk; Role; Signal Transduction; Site; Supplementation; Surface; Synapses; T cell regulation; T-Cell Receptor; T-Lymphocyte; Uridine; Variant; cell growth; glycosylation; immune function; prevent; public health relevance; src-Family Kinases; sugar

DESCRIPTION (provided by applicant): Genetic information flows from DNA to macromolecular structures; the dominant force in the molecular organization of life. However, virtually all cell surface and secreted proteins in metazoans are modified by the addition of complex carbohydrates in the ER/Golgi secretory pathway. We find that metabolite availability to the Golgi N- glycosylation pathway exerts autonomous control over the assembly of macromolecular complexes on the cell surface, and in this capacity, acts upstream of signaling and gene expression to influence cell growth, differentiation and disease states. GlcNAc branching and number of N-glycans per protein molecule (i.e. occupied N-X-S/T sites) cooperate to regulate binding to galectins and thereby the distribution, clustering and endocytosis of surface glycoproteins in a predictable manner. The number of occupied N-X-S/T sites is an encoded feature of protein sequences, while the extent of GlcNAc- branching is conditional to the enzymatic activity and metabolic supply of UDP-GlcNAc to the medial Golgi N-acetylglucosaminyltransferases I, II, IV and V (Mgat1, 2, 4 and 5). In naive T cells, galectin-3 binds the T cell receptor (TCR) and prevents spontaneous TCR oligomerization in the absence of antigen ligands. This in turn blocks recruitment of Nck, WASp, SLP-76 and CD4-Lck to TCR, F-actin remodeling and transfer of the complex to GM1/cholesterol-enriched microdomains (GEMs). This inhibits basal signaling by the Src tyrosine kinase Lck and TCR clustering and signaling at the immune synapse. The lattice also binds and localizes the tyrosine phosphatase CD45 to GEMs and the immune synapse, thereby suppressing Lck activity and TCR signaling. Once activated, membrane turnover increases and Lck/PI3K/Erk growth signaling stimulates metabolite flux to UDP-GlcNAc biosynthesis and Mgat5 activity, thereby enhancing GlcNAc branching and CTLA-4 surface retention. Mgat5 deficient 129/Sv mice develop late onset spontaneous kidney autoimmunity and are more susceptible to experimental autoimmune encephalomyelitis (EAE), an animal model of MS. EAE susceptibility among inbred mouse strains correlates inversely with GlcNAc branching in T cells. In EAE-susceptible PL/J mice, reduced GlcNAc branching is distributed across deficiencies in at least three Golgi enzymes (Mgat1, 2 and 5). Supplementing T cells with hexosamine pathway metabolites (i.e. GlcNAc, uridine) increases UDP-GlcNAc supply and GlcNAc branching, and in turn suppresses TCR hyperactivity, enhances CTLA-4 surface retention and dampens T cell mediated autoimmunity in PL/J and Non-Obese Diabetic (NOD) mice. In humans, a haplotype of MGAT1 that decreases N-glycan branching in glycoproteins by ~20% synergistically interacts with an allele of CTLA-4 that reduces N-glycan number by ~50%, increasing the risk of Multiple Sclerosis (MS) and Rheumatoid Arthritis (RA) by ~2 fold. The two variants are expected to independently reduce CTLA-4 affinity for galectins and indeed, they act cooperatively to limit CTLA-4 surface levels, a phenotype rescued by metabolic supplementation to UDP-GlcNAc biosynthesis. Taken together our data suggest that genetic and metabolic control of the galectin lattice regulates basal, activation and arrest signaling in T cells, thereby inhibiting autoimmunity in mice and humans. To extend these results, we propose three specific aims. Specific Aim 1 will further characterize critical molecules in T cells regulated by GlcNAc branching. Specific Aim 2 will explore the role of GlcNAc branching in TH17 and Treg differentiation and function. Specific Aim 3 will further define metabolic regulation of GlcNAc branching in T cells.

描述（由申请人提供）：遗传信息从DNA到大分子结构；生命分子组织中的主要力量。然而，通过在ER/Golgi分泌途径中添加复杂的碳水化合物来改变后生动物中的所有细胞表面和分泌的蛋白质。我们发现，高尔基N-糖基化途径的代谢物可利用性对大分子分子络合物组装的自主控制在细胞表面上，并以这种能力起作用信号传导和基因表达的上游，以影响细胞生长，分化和疾病状态。 GlcNAC分支和每个蛋白质分子的N-聚糖数量（即占据的N-X-S/T位点）配合以调节与半乳糖素的结合，从而以可预测的方式对表面糖蛋白的分布，聚类和内吞作用。占用的N-X-S/T位点的数量是蛋白质序列的编码特征，而GlcNAC分支的程度则是酶促活性和代谢供应UDP-GLCNAC向中内侧Golgi N-乙酰基甲基葡萄氨基转移酶I，II，II IV和V（Mgat and V（Mgat和Mgat1，2，2，4和5）。 在幼稚的T细胞中，Galectin-3结合T细胞受体（TCR），并在没有抗原配体的情况下防止自发的TCR低聚。这反过来又阻止了NCK，WASP，SLP-76和CD4-LCK的募集到TCR，F-肌动蛋白重塑和复合物向GM1/胆固醇添加的微区域（GEMS）。这抑制了SRC酪氨酸激酶LCK以及免疫突触的TCR聚类和信号传导的基础信号传导。晶格还将酪氨酸磷酸酶CD45与宝石和免疫突触结合并定位，从而抑制LCK活性和TCR信号传导。一旦激活，膜离心额就会增加，LCK/PI3K/ERK生长信号传导刺激代谢物通量到UDP-GLCNAC生物合成和MGAT5活性，从而增强了GlcNAC分支和CTLA-4表面保留。 MGAT5缺乏129/SV小鼠会发育迟发自发自身免疫性，并且更容易受到MS动物模型的实验性自身免疫性脑脊髓炎（EAE）。近交小鼠菌株之间的EAE敏感性与T细胞中的GlcNAC分支成反比。在耐Eee敏感的PL/J小鼠中，降低的GlcNAC分支分布在至少三种高尔基酶（MGAT1、2和5）的缺陷之间。用己糖胺途径代谢物（即GLCNAC，尿苷）补充T细胞会增加UDP-GLCNAC的供应和GlcNAC分支，从而抑制TCR的过度活跃性，从而增强CTLA-4表面保留率，并抑制T PLEL/J和非obese diobice（Nod）的T细胞介导的自身免疫性。在人类中，MGAT1的单倍型可将糖蛋白中N-聚糖分支降低约20％的n-聚糖分支与CTLA-4等位基因相互作用，该等位基因将N-聚糖数量降低了约50％，从而增加了多发性硬化症（MS）和Rheumatoid Althritis（RA）（RA）的风险。预计这两种变体将独立降低CTLA-4对甲状腺蛋白的亲和力，实际上，它们的作用限制了CTLA-4表面水平，这是通过补充UDP-GLCNAC生物合成的代谢而产生的表型。综上所述，我们的数据表明，对半乳肠蛋白晶格的遗传和代谢控制调节T细胞中的基础，激活和停滞信号，从而抑制小鼠和人类的自身免疫性。为了扩展这些结果，我们提出了三个具体目标。具体目标1将进一步表征受GlcNAC分支调节的T细胞中的临界分子。具体目标2将探索GlcNAC分支在TH17和Treg分化和功能中的作用。具体目标3将进一步定义T细胞中GlcNAC分支的代谢调节。

项目成果

Metabolism, cell surface organization, and disease.

• DOI: 10.1016/j.cell.2009.12.008
• 发表时间: 2009-12-24
• 期刊: Cell
• 影响因子: 64.5
• 作者: Dennis JW;Nabi IR;Demetriou M
• 通讯作者: Demetriou M

Interleukin-2, Interleukin-7, T cell-mediated autoimmunity, and N-glycosylation.

• DOI: 10.1111/j.1749-6632.2011.06391.x
• 发表时间: 2012-04
• 期刊: Annals of the New York Academy of Sciences
• 影响因子: 5.2
• 作者: Grigorian A;Mkhikian H;Demetriou M
• 通讯作者: Demetriou M

T-cell growth, cell surface organization, and the galectin-glycoprotein lattice.

• DOI: 10.1111/j.1600-065x.2009.00796.x
• 发表时间: 2009-07
• 期刊: Immunological reviews
• 影响因子: 8.7
• 作者: Grigorian A;Torossian S;Demetriou M
• 通讯作者: Demetriou M

Genetics and the environment converge to dysregulate N-glycosylation in multiple sclerosis.

• DOI: 10.1038/ncomms1333
• 发表时间: 2011
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Mkhikian, Haik;Grigorian, Ani;Li, Carey F.;Chen, Hung-Lin;Newton, Barbara;Zhou, Raymond W.;Beeton, Christine;Torossian, Sevan;Tatarian, Gevork Grikor;Lee, Sung-Uk;Lau, Ken;Walker, Erin;Siminovitch, Katherine A.;Chandy, K. George;Yu, Zhaoxia;Dennis, James W.;Demetriou, Michael
• 通讯作者: Demetriou, Michael

Mgat5 deficiency in T cells and experimental autoimmune encephalomyelitis.

• DOI: 10.5402/2011/374314
• 发表时间: 2011-01-01
• 期刊: ISRN neurology
• 作者: Grigorian, Ani;Demetriou, Michael
• 通讯作者: Demetriou, Michael