Control of Leukocyte Biology by Fucosylated Glycans

岩藻糖是一种单糖，可以修饰白细胞、内皮细胞和 上皮细胞。α-1，3岩藻糖化多糖是E-、P-和L-选择素受体拮抗剂的基本成分， 从而控制健康和疾病中的白细胞贩运。与丝氨酸和苏氨酸相连的岩藻糖 (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 发育和免疫中依赖缺口的信号转导事件。这些岩藻糖化的多糖，以及 控制它们表达的基因是治疗调节形成的关键靶点。 有助于选择素依赖的免疫和炎症的白细胞，这可能用于 基于细胞的癌症治疗。控制基因靶向缺失的小鼠的分析 α1，3岩藻糖化(Fuct-IV/VII缺失小鼠)揭示了α1，3岩藻糖化多糖在对照中的作用 关于骨髓生成的。解释这种控制的机制尚不清楚，这是 这项提议。一种允许条件性缺陷的基因(Fx)靶向缺陷小鼠的分析 普通岩藻糖化(FX缺失小鼠)推断O-岩藻糖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，也是这项提案的主要重点。 项目II的研究将部分集中于确定依赖于α1，3岩藻糖化的选择素配体 控制骨髓生成的信号转导途径，使用涉及基因表达的方法 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 Michiko)，以确定葡聚糖硫化和N-糖基化是否分别有助于 控制骨髓生成。项目II将依靠体内显微镜核心来开发时间和 空间形态与构成正常和异常骨髓生成的分子事件相关。 项目二还将研究Notchl的O-岩藻糖化在控制骨髓生成中的作用，使用Notchendate 骨髓细胞分析和针对0-岩藻糖化多肽的抗体(来自合成 核心)。项目二还将对重组Notchl和Notch配体使用原子力显微镜方法 确定Notchl与其配体之间的相互作用如何被Olink调节的分子基础 岩藻糖，以及修饰O-连接岩藻糖的多糖。

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.