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: 8308589
• 负责人: JOHN B LOWE
• 金额: $ 41.65万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8308589
• 关键词: Accounting; Adhesions; Antibodies; Atomic Force Microscopy; Biological Assay; Biology; Carbohydrates; Cells; Characteristics; Collaborations; Development; Disease; EGF gene; Endothelial Cells; Epithelium; Event; Fucose; Gene Expression; Genes; Health; Immunity; Inflammation; Knockout Mice; Leukocyte Trafficking; Leukocytes; Ligands; Link; Methods; Molecular; Monosaccharides; Mus; Myelopoiesis; Normal Cell; Peptide Synthesis; Polysaccharides; Proteomics; Recombinants; Regulation; Role; Selectins; Serine; Signal Transduction; Signal Transduction Pathway; Threonine; Work; base; cancer therapy; glycosylation; intravital microscopy; neoplastic cell; notch protein; receptor; sulfation; therapeutic target

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，其修饰白细胞、内皮细胞和血管内皮细胞上的N-和O-连接聚糖。 上皮细胞α 1，3岩藻糖基化聚糖是E-、P-和L-选择素的反受体的基本组分， 从而控制健康和疾病中的白细胞运输。与丝氨酸和苏氨酸连接的岩藻糖 （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 Notch-dependent signal transduction events in development and immunity.这些岩藻糖基化聚糖，以及 控制其表达的基因代表了治疗性调节形成的关键靶点 白细胞，有助于选择素依赖性免疫和炎症，并可用于 基于细胞的癌症治疗对具有控制基因的靶向缺陷的小鼠的分析 α 1，3岩藻糖基化（FucT-IV/VII缺失小鼠）公开了α 1，3岩藻糖基化聚糖在对照中的作用。 骨髓增生解释这种控制的机制尚不清楚，这也是本报告的主要重点。 这个提议。对具有允许条件性缺陷的基因（FX）靶向缺陷的小鼠的分析 一般岩藻糖基化（FX无效小鼠）的影响推断O-岩藻糖基化在骨髓生成控制中的作用。 目前尚不清楚这种控制的机制，这也是本提案的一个主要重点。 项目II的研究将部分集中于鉴定α 1，3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。这项工作将包括与项目I（福田实博士）的合作 和III（Fukuda），以确定聚糖硫酸化和N-糖基化是否分别有助于 控制骨髓生成。项目II将依靠活体显微镜核心， 空间形态学与正常和异常骨髓生成的分子事件相关。 项目II还将使用Notchdependent方法研究Notchl的O-岩藻糖基化在控制骨髓生成中的作用。 骨髓生成测定和对0-岩藻糖基化肽特异性的抗体（来自合成 核心）。项目II还将使用原子力显微镜方法与重组Notchl和Notch配体 为了定义Notchl及其配体之间的相互作用如何被Olinked调节的分子基础， 岩藻糖和修饰O-连接岩藻糖的聚糖。