MODULATION OF INTEGRIN FUNCTION BY CHANGES IN N-LINKED OLIGOSACCHARIDE BRANCHING

描述（由申请人提供）：糖蛋白和糖脂在人体生理学和疾病中具有许多功能，已知这些细胞表面分子上聚糖的表达在早期胚胎发育期间发生变化。胚胎干细胞来源于早期胚胎，可以在体外培养并诱导分化为多种细胞类型。控制这种分化和产生分化干细胞的同质群体是生物医学界的主要挑战。发展细胞标记物和技术来丰富细胞群可以大大有助于干细胞研究。我们建议建立一个生物医学糖组学综合资源中心，该中心将专注于开发工具和技术，以详细分析小鼠胚胎干细胞及其分化细胞的糖蛋白和糖脂表达。项目1，胚胎干细胞计划，将开发大量培养干细胞的工具，并详细研究在特定细胞表面糖蛋白上表达的聚糖。项目2将开发技术，以分离干细胞产生的糖肽，并确定表达这些聚糖的亲本蛋白质以及这些聚糖在哪些位点表达。项目3将开发动态实时PCR来研究糖基转移酶转录本的表达。项目4将开发生物信息学工具，将结构和表达数据输入关系数据库进行处理和分析。本资源中开发的技术将使人们了解糖基化在分化过程中是如何控制的，并将允许开发工具来促进干细胞治疗人类疾病。此外，该技术将适用于其他细胞类型的研究，包括正在发展为更具侵袭性表型的癌细胞。这一糖组学资源将导致技术发展，使科学界的其他人能够通过传播开发的新方法和通过资源向其他科学家提供的分析服务参与糖组学研究。还将向科学界提供关于已开发技术的培训课程。合作项目已经构成了大量的拟议研究，并将成为开发糖组学资源的一个越来越重要的组成部分。

DESCRIPTION (provided by applicant): Glycoproteins and glycolipids have many functions in human physiology and disease and the expression of the glycans on these cell surface molecules are known to change during early embryonic development. Embryonic stem cells, derived from early embryos, can be grown in vitro and induced to differentiate into many cell types. Controlling this differentiation and producing homogeneous populations of differentiated stem cells are major challenges for the biomedical community. Development of cell markers and technology to enrich cell populations could aid significantly stem cell research. We propose to establish an Integrated Resource Center for Biomedical Glycomics that will focus on developing tools and technology to analyze in detail the glycoprotein and glycolipid expression of mouse embryonic stem cells and cells into which they differentiate. Project 1, Embryonic Stem Cell Program, will develop the tools to culture large numbers of stem cells and investigate in detail the glycans expressed on specific cell surface glycoproteins. Project 2 will develop technology to fractionate glycopeptides produced from stem cells and determine the parent proteins that express these glycans and at which sites these glycans are expressed. Project 3 will develop kinetic real-time PCR to study the expression of glycosyltransferase transcripts. Project 4 will develop Bioinformatic tools to input structural and expression data into relational databases for processing and analysis. The technology developed in this Resource will allow an understanding of how glycosylation is controlled during differentiation and will allow the development of tools to promote the use of stem cells to treat human disease. In addition, the technology will be applicable to the study of other cell types, including cancer cells that are progressing to a more invasive phenotype. This glycomics resource will result in technology development that will allow others in the scientific community to participate in glycomics research via dissemination of the new methods developed and via analytical service provided by the resource to other scientists requesting help in glycomic analyses. Training courses in the developed technologies will also be provided to the scientific community. Collaborative projects already constitute a significant amount of the proposed research and will become an increasingly important component of the developing glycomics resource.