SELECTIN MEDIATED CELL ADHESION UNDER HYDRODYNAMIC SHEAR

选择流体力学剪切下介导的细胞粘附

基本信息

批准号：
7819181
负责人：
SRIRAM NEELAMEGHAM
金额：
$ 2.05万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-08 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7819181
关键词：
2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole Acute Address Adhesions Affect Affinity Anabolism Area Binding Biochemical Biochemistry Biological Assay Blood Vessels Carbohydrates Cardiovascular Diseases Cell Adhesion Cell Adhesion Molecules Cell Line Cell surface Cells Chemicals Chimera organism Complement Complex Data Development Disaccharides Disease Dose Drug Design Engineering Epitopes Family Family member Figs - dietary Flow Cytometry Fucose Future Glycolipids Glycoproteins HL60 Human Immune response Infiltration Inflammation Inflammatory Kinetics Lead Lectin Leukocytes Ligand Binding Ligands Link Liquid Chromatography Measurement Measures Mediating Membrane Glycoproteins Metabolic Metabolic Pathway Methods Modeling Modification Molecular Biology Monosaccharides O-Glycans Biosynthesis Pathway Oligosaccharides P-Selectin P-selectin ligand protein Pathology Pathway interactions Peritonitis Physiological Polysaccharides Process Proteins Reaction Reagent Reporting Research Personnel Research Project Grants Running Selectins Simulate Source Structure Surface Plasmon Resonance Systems Biology Testing Thioglycolates Tissues Trisaccharides Ursidae Family Vascular Diseases Vascular Endothelial Cell Western Blotting Work analog base carbohydrate binding protein carbohydrate receptor carbohydrate structure chemical synthesis combat data modeling design feeding fluid flow glycosylation glycosyltransferase in vivo inhibitor/antagonist insight interest mathematical model mouse model novel numb protein prevent programs reaction rate research study small molecule sulfated glycoprotein p50 tool

项目摘要

This research project examines the function of carbohydrate chains that are O-linked to leukocyte cell- surface glycoproteins. By acting as the natural ligands of the selectin family of adhesion molecules, these glycoproteins control the rates of leukocyte adhesion in the human vasculature during normal immune response, inflammatory diseases and certain types of cardiovascular disorders. It is widely believed that controlling the rate of leukocyte adhesion in vascular disorders can lead to new therapies to combat these ailments. Thus, in the current proposal, we evaluate two mechanisms for controlling selectin-ligand binding. In Aim 1, we develop and test the ability of unique molecules based on an unusual disaccharide carbohydrate structure (GalNAc(31,3GalNAca-O-Methyl) to competitively inhibit selectin binding interactions with its ligand. Our preliminary data suggests that this disaccharide alone can bind P- selectin. We also demonstrate that appropriate modification of this unit can dramatically enhance the binding affinity of the resulting carbohydrate for selectins, when compared with the prototypic selectin ligand sialyl Lewis-X. In Aim 2, we test an approach where small-molecule metabolic inhibitors are designed based on the structure of monosaccharides that compose natural selectin ligands. These modified monosaccharidesare fed to cells in order to interfere with the biosynthesis of specific carbohydrate epitopes on the glycoprotein ligands of selectins. More specifically, these molecules are directed to alter either the core or terminal residues of glycans expressed by an important leukocyte selectin-ligand called PSGL-1 (P-selectin glycoprotein ligand- 1). We evaluate the ability and mechanism by which these chemical inhibitors permeate cells, engage and modify glycan biosynthetic pathways and inhibit cell adhesion. In Aim 3, to complement the experimental work above, a Systems Biology based mathematical model is developed to simulate biochemical networks that regulate O-glycan biosynthesis in leukocytes. Many of the assumptions in this mathematical model are experimentally validated. Diverse experimental methods are applied to accomplish the above three aims. These include cell adhesion studies under controlled flow, in vivo experiments in a mouse model of acute inflammation, western blot analysis, molecular biology based approaches, flow cytometry, surface plasmon resonance and liquid chromatography. In the long run, we anticipate that small-molecule selectin-antagonists will be identified from this work that may aid future drug design. Mathematical models developed will enhance the application of metabolic engineering principles in the area of biological chemistry. Such analysis can also provide the rationale for the chemical synthesis of new inhibitors and for interpretation of experimental observations.

该研究项目研究了碳水化合物链的功能，这些链条与白细胞细胞的O连接表面糖蛋白。通过充当粘附分子选择素家族的天然配体，这些糖蛋白控制正常免疫期间人血管中白细胞粘附的速率反应，炎症性疾病和某些类型的心血管疾病。人们普遍相信控制血管疾病中白细胞粘附的速率可能会导致新疗法与这些疗法作斗争疾病。因此，在当前建议中，我们评估了控制选择素 - 配体结合的两种机制。在AIM 1中，我们基于异常二糖来发展和测试独特分子的能力碳水化合物结构（galnac（31,3galnaca-o-甲基）以竞争性抑制选择素结合相互作用带有配体。我们的初步数据表明，仅这种二糖可以结合p-selectin。我们也是证明对本单元的适当修改可以显着增强与原型选择配体siallyl lewis-x相比，选择蛋白的碳水化合物产生的碳水化合物。目标 2，我们测试一种方法，该方法是根据基于结构的结构而设计的小分子代谢抑制剂构成天然选择配体的单糖。这些改良的单糖饲料喂入细胞为了干扰特定碳水化合物表位的生物合成在糖蛋白配体上 Selectins。更具体地说，这些分子被指示以改变由重要白细胞选择素配体表达的聚糖称为PSGL-1（P-选择素糖蛋白配体 - 配体 - 1）。我们评估了这些化学抑制剂渗透细胞，参与和的能力和机制修饰聚糖生物合成途径并抑制细胞粘附。在AIM 3中，以补充实验上面的工作，开发了基于系统生物学的数学模型来模拟生化网络调节白细胞中的O-聚糖生物合成。这个数学模型中的许多假设是经过实验验证。采用各种实验方法来实现上述三个目标。这些包括在受控流动下的细胞粘附研究，急性小鼠模型中的体内实验炎症，蛋白质印迹分析，基于分子生物学的方法，流式细胞仪，表面等离子体共振和液相色谱。从长远来看，我们预计小分子选择素 - 将从这项工作中确定可能有助于未来的药物设计。开发的数学模型将增强代谢工程原理在生物化学领域的应用。这样的分析还可以提供新抑制剂化学合成的理由和解释实验观察。