Control of Leukocyte Biology by Fucosylated Glycans

岩藻糖基化聚糖对白细胞生物学的控制

• 批准号: 7534123
• 负责人: JOHN B LOWE
• 金额: $ 42.96万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7534123
• 关键词: Accounting; Acute; Address; Antibodies; Apoptosis; Atomic Force Microscopy; Binding; Biological Assay; Biology; Birth; Cell Proliferation; Cells; Characteristics; Collaborations; Complex; Couples; Development; Disabled Persons; Disease; Endothelial Cells; Enzymes; Epithelium; Event; Fuc-TIV; Fuc-TVII; Fucose; Fucosyltransferase; Gene Expression; Generations; Genes; Health; Hematopoietic; Homeostasis; Homing; Image; Immunity; In Vitro; Inflammation; Inorganic Sulfates; Knockout Mice; Lead; Leukocyte Trafficking; Leukocytes; Life; Ligands; Link; Literature; Location; Lymphopoiesis; Mannosidase; Marrow; Methods; Molecular; Monoclonal Antibodies; Monosaccharides; Mus; Myelogenous; Myeloid Progenitor Cells; Myelopoiesis; P-Selectin; Pathway interactions; Peptide Synthesis; Phenotype; Polysaccharides; Process; Proteins; Proteomics; Rate; Reagent; Recombinants; Regulation; Relative (related person); Research Personnel; Role; Selectins; Serine; Signal Transduction; Signal Transduction Pathway; Site; Specificity; Stem cells; Structure; Surface Plasmon Resonance; System; Testing; Thinking; Threonine; Time; Unspecified or Sulfate Ion Sulfates; Wild Type Mouse; Work; base; cancer therapy; embryonic stem cell; glycosylation; granulocyte; in vivo; intravital microscopy; monocyte; mutant; neutrophil; notch protein; progenitor; programs; receptor; reconstitution; research study; sulfation; therapeutic target; tool

Fucose is a monosaccharide that modifies N- and O-linked glycans on leukocytes, endothelial cells and epithelia. Alpha1,3fucosylated glycans are essential components of the counter-receptors for E-, P-, and Lselectin, and thus control leukocyte trafficking in health and disease. Fucose linked to serine and threonine (O-linked fucose) is characteristic of some EGF-like repeats of Notch and its ligands, and contributes to Notch-dependent signal transduction events in development and immunity. These fucosylated glycans, and the genes that control their expression represent pivotal targets for therapeutic regulation of the formation of leukocytes that contribute to selectin-dependent immunity and inflammation, and that may be used for cell-based treatment of cancer. Analyses of mice with targeted deficiencies in genes that control alpha1,3fucosylation (FucT-IV/VII-null mice) disclose a role for alpha1,3fucosylated glycans in the control of myelopoiesis. Mechanisms to account for such control are not yet known, and are a primary focus of this proposal. Analyses of mice with a targeted deficiency in a gene (FX) that allows conditional deficiency of general fucosylation (FX null mice) infer a role for O-fucosylation in the control of myelopoiesis. Mechanisms to account for such control are not yet known, and are also a primary focus of this proposal. Studies in Project II will focus in part on identifying the alpha1,3fucosylated selectin ligand-dependent signal transduction pathways that control myelopoiesis, using approaches that involve gene expression arrays and proteomics analyses. This work will include collaborations with Projects I (Dr. Minoru Fukuda) and III (Michiko Fukuda) to determine if glycan sulfation and N-glycosylation, respectively, contribute to the control of myelopoiesis. Project II will rely on the Intravital Microscopy Core to develop temporal and spatial morphological correlates for the molecular events that underlie normal and aberrant myelopoiesis. Project II will also study the role of O-fucosylation of Notchl in controlling myelopoiesis, using Notchdependent myelopoietic assays, and antibodies specific for 0-fucosylated peptides (from the Synthesis Core). Project II will also use atomic force microscopy methods with recombinant Notchl and Notch ligands to define the molecular basis for how the interactions between Notchl and its ligands are modulated by Olinked fucose, and by the glycans that modify O-linked fucose.

岩藻糖是一种单糖，可修饰白细胞、内皮细胞和 上皮细胞。 Alpha1,3 岩藻糖基化聚糖是 E-、P- 和 L 选择素反受体的重要组成部分， 从而控制健康和疾病中的白细胞贩运。岩藻糖与丝氨酸和苏氨酸连接 （O-连接岩藻糖）是 Notch 及其配体的一些 EGF 样重复序列的特征，并有助于 发育和免疫中Notch依赖性信号转导事件。这些岩藻糖基化聚糖，以及 控制其表达的基因代表了形成治疗调节的关键靶点 有助于选择素依赖性免疫和炎症的白细胞，可用于 基于细胞的癌症治疗。对控制基因有针对性缺陷的小鼠进行分析 α1,3 岩藻糖基化（FucT-IV/VII-null 小鼠）揭示了 α1,3 岩藻糖基化聚糖在对照中的作用 骨髓生成。解释这种控制的机制尚不清楚，并且是主要关注点 这个建议。对具有允许条件性缺陷的基因（FX）目标缺陷的小鼠进行分析 一般岩藻糖基化（FX 无效小鼠）的研究推断 O-岩藻糖基化在控制骨髓生成中的作用。 解释这种控制的机制尚不清楚，这也是该提案的主要焦点。 项目 II 的研究将部分集中于鉴定 α1,3 岩藻糖基化选择素配体依赖性 使用涉及基因表达的方法控制骨髓生成的信号转导途径 阵列和蛋白质组学分析。这项工作将包括与 Projects I（福田稔博士）的合作 和 III (Michiko Fukuda) 分别确定聚糖硫酸化和 N-糖基化是否有助于 控制骨髓生成。项目 II 将依靠活体显微镜核心来开发时间和 正常和异常骨髓生成​​的分子事件的空间形态学相关性。 项目II还将利用Notchdependent研究Notch1的O-岩藻糖基化在控制骨髓生成中的作用 骨髓生成测定和 0-岩藻糖基化肽特异性抗体（来自 Synthesis 核）。项目 II 还将使用重组 Notch1 和 Notch 配体的原子力显微镜方法 定义 Olinked 如何调节 Notch1 及其配体之间相互作用的分子基础 岩藻糖，以及修饰 O-连接岩藻糖的聚糖。