STRUCTURE AND BIOSYNTHESIS OF SELECTIN LIGANDS

选择素配体的结构和生物合成

• 批准号: 6573075
• 负责人: MINORU FUKUDA
• 金额: $ 26.5万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-15 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6573075
• 关键词: Peyer's patches; carbohydrate biosynthesis; carbohydrate structure; cell membrane; cytolysis; enzyme mechanism; gene expression; genetically modified animals; glycoproteins; glycosyltransferase; high endothelial venule; human tissue; laboratory mouse; ligands; lymph nodes; metastasis; molecular cloning; natural killer cells; neoplasm /cancer; oligosaccharides; selectins; sialyltransferases; sulfotransferase; tissue /cell culture; transfection

Project I- Structure and Biosynthesis of Selectin Ligands (Minoru Fukuda, Ph.D.) In the previous funding period, we have made critical progress in this field. First, we discovered that 6-sulfo sialyl Lewis x in high endothelial venules (HEV) can be formed in core2 branch and extended core1 O- glycans. Expression of novel sulfotransferase (LSST) and core1 extension enzyme (core1-beta3GlcNAcT) demonstrated that the expression of these structures in biantennary O-glycans leads to highly active L-selectin ligands. Second, that excessive expression of sialyl Lewis x on tumor cells results in cytolysis of those tumor cells by nature killer (NK) cells, while moderate expression of sialyl Lewis x facilitates tumor metastasis. Based on these findings, two major areas of study are proposed: I. Elucidation of structure and function of L-selectin ligands in HEV at peripheral lymph nodes and Peyer's patches. In collaboration with Drs. John Lowe, Michiko Fukuda, and Ole Hindsgaul, we will determine the complete structure of L-selectin ligands at HEV in peripheral and mucosal lymph nodes, and determine the roles of 6-sulfo sialyl Lewis x is an L-selectin ligand by generating a mutant mouse. II. Roles of carbohydrates in tumor metastasis. In collaboration with Drs. Michiko Fukuda and John Lowe, we will determine if the amount of sialyl Lewis x plays a major role in facilitating tumor metastasis or NK cell-mediated cytolysis. We will also determine if cell surface carbohydrates isolated from tumor cells inhibit tumor metastasis or NK cell-mediated cytolysis. These studies will allow us to understand the structure and biosynthesis of selectin ligands and the roles of cell surface carbohydrates of tumor cells in tumor metastasis and immune cell recognition.

项目 I——选择素配体的结构和生物合成（Minoru Fukuda，博士） 在上一个资助期间，我们在这一领域取得了关键进展。首先，我们发现高内皮微静脉 (HEV) 中的 6-磺基唾液酸路易斯 x 可以在 core2 分支和延伸的 core1 O-聚糖中形成。新型磺基转移酶 (LSST) 和 core1 延伸酶 (core1-beta3GlcNAcT) 的表达表明，这些结构在双触角 O-聚糖中的表达可产生高活性的 L-选择素配体。其次，肿瘤细胞上唾液酸Lewis x 的过度表达导致这些肿瘤细胞被自然杀伤（NK）细胞溶解，而唾液酸Lewis x 的适度表达则促进肿瘤转移。基于这些发现，提出了两个主要研究领域： I. 阐明外周淋巴结和派尔氏集结处 HEV 中 L-选择素配体的结构和功能。与博士合作。 John Lowe、Michiko Fukuda 和 Ole Hindsgaul，我们将确定外周和粘膜淋巴结中 HEV 处 L-选择素配体的完整结构，并通过生成突变小鼠。二.碳水化合物在肿瘤转移中的作用。与博士合作。 Michiko Fukuda 和 John Lowe，我们将确定唾液酸 Lewis x 的量是否在促进肿瘤转移或 NK 细胞介导的细胞溶解中发挥主要作用。我们还将确定从肿瘤细胞中分离的细胞表面碳水化合物是否抑制肿瘤转移或 NK 细胞介导的细胞溶解。这些研究将使我们了解选择素配体的结构和生物合成以及肿瘤细胞的细胞表面碳水化合物在肿瘤转移和免疫细胞识别中的作用。