Mechanisms and functions of cell surface glycoRNAs

细胞表面糖RNA的机制和功能

• 批准号: 10712185
• 负责人: Ryan Alexander Flynn
• 金额: $ 44.25万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-14 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10712185
• 关键词: Anabolism; Biochemical; Biogenesis; Biological Assay; Biology; Biophysics; Caenorhabditis elegans; Carbohydrates; Carboxylic Acids; Cell Communication; Cell model; Cell physiology; Cell surface; Cells; Cellular biology; Chemicals; Communities; Complex; Diagnostic; Dissection; Engineering; Environment; Eukaryota; Exclusion; Fission Yeast; Genetic; Glycobiology; Hybrids; Image; Intracellular Space; Label; Link; Mammalian Cell; Metabolic; Methods; Modality; Molecular; Nature; Neighborhoods; Organism; Pathogenicity; Pathway interactions; Play; Polymers; Polysaccharides; Positioning Attribute; Prokaryotic Cells; RNA; Receptor Signaling; Regulation; Reporter; Role; Saccharomyces cerevisiae; Signal Transduction; Surface; Therapeutic; Therapeutic Intervention; Work; Yeasts; cell type; field study; glycosylation; immunoregulation; innovation; novel; novel strategies; pathogen; receptor; sialic acid binding Ig-like lectin; tool; tool development

PROJECT SUMMARY/ ABSTRACT The cell surface is a platform for physical and regulatory control over cell biology, positioning it to be a key interface for diagnostic targeting and therapeutic intervention. While RNA is a central polymer in biology most thought and experimental effort devoted to RNA biology has been confined to intracellular spaces and excluded from participating in cell surface biology. On the cell surface, carbohydrate polymers (glycans) are of critical importance due to biophysical and signaling activities. Interestingly, despite both polymers playing central roles in biology, RNA and glycans have largely existed in entirely non-overlapping fields of study. However, my work has provided evidence of a hybrid molecule, an RNA-glycan conjugate (glycoRNA); this new class of biomolecule represents a direct link between RNA and glycobiology. Critically, glycoRNAs are localized to the external surface of living cells and can engage with immunomodulatory Siglec receptors. Thus, glycoRNAs are positioned on a surface of critical regulatory importance, with access to cell-cell interactions, pathogens, and signaling receptors on the cell surface. However, we currently lack facile tools to study this new cell surface molecule, we do not understand the molecular or atomic composition of glycoRNAs, and we have a poor understanding of how many species biosynthesize glycoRNAs. This MIRA proposal is focused on developing and implementing methods to uncover functional roles of RNA glycosylation and we will approach the complex biology of glycoRNAs in a systematic fashion. Initially we will develop novel chemical approaches to label glycans in the context of RNA. My proposed strategy of selective carboxylic acid labeling represents an innovative new approach to detecting glycoRNA, without the need for synthetic metabolic reporters. These tools will be easily implemented across cell types and species enabling others in the scientific community. We will apply these tools and other molecular assays to expand our understanding of the composition of the cell surface in the context of glycoRNA. Biochemical, biophysical, and imaging-based strategies will be used to define the molecular neighborhoods of glycoRNAs as well as the chemical nature of the RNA-glycan linkage; all together providing a more complete picture of the mammalian cell surface. Finally, we will develop the first evidence of glycoRNAs in non-mammalian organisms. First focusing on two major strains of yeast (S. cerevisiae and S. pombe) with robust culturing, functional, and genetic tools that will allow for rapid dissection of the biogenesis pathway for eventual engineering purposes. Expanding to other organisms including prokaryotes (pathogenic and not) as well as other multicellular eukaryotes like C. elegans will better define the scope of glycoRNA biosynthesis and more robustly equip us to generate synthetic glycoRNAs. More broadly, we intend to advance the general model of how cells interact with each other, pathogens, and exogenous molecules, as without cell surface glycoRNAs, they are likely incomplete. This proposal develops innovative methods to establish new conceptual and physical layers of regulation between a cell and its environment.

项目摘要/摘要 细胞表面是对细胞生物学的物理和调节控制的平台，将其定位为关键 诊断靶向和治疗干预的接口。虽然RNA是生物学的中心聚合物 专门用于RNA生物学的思想和实验努力已局限于细胞内空间，排除在外 参与细胞表面生物学。在细胞表面，碳水化合物聚合物（聚糖）很关键 由于生物物理和信号活性而引起的重要性。有趣的是，尽管两个聚合物都扮演着核心角色 在生物学中，RNA和Glycans在很大程度上存在于完全非重叠的研究领域中。但是，我的工作 提供了混合分子，RNA - 聚糖偶联物（Glycorna）的证据；这类新的生物分子 代表RNA与糖生物学之间的直接联系。至关重要的是，糖原位于外部 活细胞的表面并可以与免疫调节的Siglec受体接合。因此，糖原被定位 在临界调节重要性的表面上，可以访问细胞 - 细胞相互作用，病原体和信号传导 细胞表面的受体。但是，我们目前缺乏研究这个新细胞表面分子的轻松工具，我们 不了解糖原的分子或原子成分，我们对 有多少物种生物合成糖原。 MIRA提案的重点是开发和实施 发现RNA糖基化功能作用的方法，我们将接近复杂的生物学 以系统的方式进行聚糖。最初，我们将开发新颖的化学方法来将糖标记在 RNA的上下文。我提出的选择性羧酸标签的策略代表了一种创新的新型 无需合成代谢记者而无需检测糖性的方法。这些工具很容易 跨细胞类型和物种实施，在科学界实现了其他人。我们将应用这些工具 和其他分子测定，以扩大我们对细胞表面组成的理解 Glycorna。生化，生物物理和基于成像的策略将用于定义分子 Glycornas的社区以及RNA-Glycan连接的化学性质；一起提供 哺乳动物细胞表面的更完整图片。最后，我们将在 非哺乳动物生物。首先专注于两种主要的酵母菌菌株（S. cerevisiae和S. pombe） 培养，功能和遗传工具将允许生物发生途径快速解剖以最终 工程目的。扩展到包括原核生物（病原体和非原核生物）以及其他生物 诸如秀丽隐杆线虫之类的多细胞真核生物将更好地定义糖原生物合成的范围，更牢固地定义 设备我们生成合成的聚糖。更广泛地，我们打算推进细胞的一般模型 彼此相互作用，病原体和外源分子，例如没有细胞表面糖菌，它们是 可能不完整。该建议开发了创新方法来建立新的概念和物理层 细胞及其环境之间的调节。