Function and regulation of epithelial glycosylation

上皮糖基化的功能和调节

• 批准号: 10621189
• 负责人: Jennifer J Kohler
• 金额: $ 52.57万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10621189
• 关键词: Binding; Biological; Biology; CRISPR screen; Candidate Disease Gene; Cell surface; Cells; Chemicals; Cholera; Cholera Toxin; Collaborations; Communicable Diseases; Development; Disease; Epithelium; Ganglioside GM1; Genes; Glycoconjugates; Goals; Human; Individual; Intoxication; Length; Malignant Neoplasms; Mammalian Cell; Methods; Molecular Structure; Physiology; Polysaccharides; Predisposition; Production; Reagent; Regulation; Research; Role; Shapes; Surface; Universities; airway epithelium; analog; choleragen receptor; crosslink; design; experimental study; glycosylation; host-microbe interactions; interest; intestinal epithelium; novel; programs; receptor; sugar; tool

Summary/abstract This research program focuses on uncovering the biological roles of glycoconjugates. Part of this effort is devoted to the development of chemical biology tools for glycoscience research. In the past, we have developed photocrosslinking sugar analogs that can be incorporated into cellular glycoconjugates and used to covalently crosslink glycoconjugates to their binding partners in a native context. These reagents can be used to identify glycan-dependent binding interactions, and to characterize where and under what conditions that these interactions occur. Over the next five years, we will further expand the scope of experiments that can be performed by preparing additional photocrosslinking sugars, developing new methods for their incorporation, and evaluating their incorporation into additional glycoconjugates. Using one of these photocrosslinking sugars, we made the unexpected observation that cholera toxin can bind fucosylated glycoconjugates in addition to its canonical receptor, the ganglioside GM1. Over the next five years, we will determine the molecular structure of fucosylated glycoconjugates recognized by cholera toxin and characterize their role in host cell intoxication. These studies are supported by our long-term collaboration with the Yrlid group (University of Gothenburg) and their expertise in studying cholera disease mechanisms. Our studies of cholera toxin receptors led us to become interested in the diverse glycoconjugates that line the intestinal and respiratory epithelia. A CRISPR screen designed to identify genes that modulate cholera toxin binding to cell surfaces identified a number of candidate genes that may function in the regulation of glycosylation by diverse mechanisms. Over the next five years, we will characterize novel regulators of glycosylation and determine how they shape the glycome, modulating glycan features such as polyLacNAc chain length and the degree of fucosylation. The long-term goal of these studies is to determine how glycan features vary among individuals, their association with disease states, and their impact on host-microbe interactions.

总结/摘要 该研究项目的重点是揭示糖复合物的生物学作用。这一努力的一部分是 致力于开发用于糖科学研究的化学生物学工具。在过去，我们发展了 光交联糖类似物，其可掺入细胞糖缀合物中并用于共价结合 将糖缀合物与它们在天然环境中的结合配偶体交联。这些试剂可用于鉴定 聚糖依赖性结合相互作用，并表征在何处和在何种条件下，这些 互动发生。在未来五年，我们将进一步扩大实验范围， 通过制备额外的光交联糖，开发新的掺入方法， 并评价它们掺入另外的糖缀合物中。利用其中一种光交联 糖，我们意外地观察到霍乱毒素可以结合岩藻糖基化糖缀合物， 除了它的典型受体，神经节苷脂GM 1。在未来五年，我们将确定 被霍乱毒素识别的岩藻糖基化糖缀合物的分子结构，并表征它们在 宿主细胞中毒这些研究得到了我们与Yrlid集团长期合作的支持 （哥德堡大学）和他们在研究霍乱疾病机制方面的专业知识。我们对霍乱的研究 毒素受体使我们对排列在肠道和胃肠道周围的各种糖复合物产生了兴趣， 呼吸道上皮。CRISPR筛选旨在鉴定调节霍乱毒素与细胞结合的基因 表面鉴定了许多候选基因，这些基因可能通过多种途径调节糖基化。 机制等在接下来的五年里，我们将描述新型糖基化调节剂的特征，并确定其作用机制。 它们形成糖组，调节聚糖特征，如聚LacNAc链长和糖链的程度。 岩藻糖化这些研究的长期目标是确定聚糖特征如何在个体之间变化， 它们与疾病状态的关联，以及它们对宿主-微生物相互作用的影响。