Systems Biology of Glycosylation

这是一个补助金，已被支持作为NHLBI系统生物学计划的一部分更新应用。 它涉及的主题是糖基化，一个普遍存在的和复杂的翻译后修饰在哺乳动物 细胞聚糖修饰绝大多数哺乳动物蛋白质。他们绝对控制或微调了一些 高等生物的细胞过程包括发育、免疫、炎症、出血和转移。 聚糖结构的改变是由于细胞代谢、发育和信号传导事件的改变， 潜在的转录组系统地了解控制因素（遗传和表观遗传） 糖基化目前不可用。然而，这是重要的，原因有二：一。这些知识可以帮助 在不同的细胞系统之间建立定量联系，以便在一个系统的研究中获得的知识 可以用来预测另一个世界的结果。二.基因转录测量现在被用于临床 新一代测序（NGS）的出现大大降低了这种诊断的成本。 测定。如果基因表达模式的变化和聚糖结构的改变之间的关系是 可以使用标准基因测序方法来测定已建立的疾病相关聚糖生物标志物。 这可以在精准医疗应用期间实现早期诊断和患者分层。基于 如上所述，当前的提议解决了“基于定量， 分析实验与模型的建立可以帮助相关细胞糖组学的变化， 潜在的转录组”。这一命题将通过一系列使用血液的研究来检验。 白细胞参与先天性免疫，并通过将结果与炎性白细胞-内皮细胞 粘附力学具体目标是：1.使用单细胞分析开发血液聚糖图谱， NGS平台和基于糖探针的质谱法。这一目标将导致一个关系数据库， 描述了调节三种骨髓/单核细胞中特定聚糖结构生物合成的糖酶 血细胞系、原代人血和CD 34+造血干/祖细胞（HSPC）。2：发展 一个互补的实验建模框架，将糖基因表达与聚糖结构联系起来。这一目标 扩展了目标1中的概念，仅关注正在分化为中性粒细胞或 巨噬细胞谱系。新兴数据将产生识别新控制器的糖原调控网络图 细胞的糖基化特征。3：测试所选小分子、转录因子（TF）和 在离体和体内白细胞-内皮细胞粘附过程中的糖基因检查点。在这里，我们确定 数据驱动的计算机预测和分子研究的能力，以确定新的检查点调节人类 炎性白细胞粘附。总的来说，这项研究从分子到细胞和整个动物水平。 它将产生最先进的系统生物学计算和实验工具，以提高我们的 了解细胞糖基化、血液聚糖及其对人类炎症性疾病的影响。

This is the renewal application of a grant that has been supported as part of the NHLBI Systems Biology program. It deals with the subject of glycosylation, a ubiquitous and complex post-translational modification in mammalian cells. Glycans decorate a vast majority of mammalian proteins. They absolutely control or fine-tune a number of cellular processes in higher organisms including development, immunity, inflammation, bleeding and metastasis. Glycan structures change due to alterations in cell metabolism, development and signaling events that perturb the underlying transcriptome. A systematic understanding of the factors (genetic and epigenetic) controlling glycosylation is currently unavailable. This is however important for two reasons: i. Such knowledge can help establish quantitative links between different cell systems, so that knowledge gained in the study of one system can be applied to predict the outcome in another. ii. Gene transcript measurements are now being used in clinical diagnostics, and the advent of next generation sequencing (NGS) has dramatically reduced the cost of such assays. If a relation between changes in the pattern of gene expression and alterations in glycan structures is established, disease-associated glycan biomarkers may be assayed using standard gene sequencing methods. This can enable both early diagnosis and patient stratification during precision medicine applications. Based on the above, the current proposal addresses the hypothesis that “Coupling systems based quantitative, analytical experimentation with model building can help relate cellular glycomics changes to the underlying transcriptome”. This proposition will be tested by performing a series of studies using blood leukocytes involved in innate immunity and by relating findings to inflammatory leukocyte-endothelial cell adhesion mechanics. The specific aims are: 1. To develop a blood glycan atlas using single-cell analysis on NGS platform and glycoProbe based mass spectrometry. This aim will result in a relational database that describes the glycoEnzymes regulating the biosynthesis of specific glycan structures in three myeloid/monocytic blood cell lines, primary human blood and in CD34+ hematopoietic stem/progenitor cells (HSPC). 2: To develop a complementary experiment-modeling framework to relate glycogene expression to glycan structure. This aim extends concepts in Aim 1, only focusing on blood cells that are being differentiated down neutrophil or macrophage lineages. Emerging data will yield glycogene regulatory network maps that identify novel controllers of cellular glycosylation profile. 3: To test the roles of selected small molecules, transcription factors (TFs) & glycogene checkpoints during leukocyte-endothelial cell adhesion ex vivo and in vivo. Here, we determine the ability of data-driven computer predictions and molecular studies, to identify new checkpoints regulating human inflammatory leukocyte adhesion. Overall, the study scales from the molecular, to cell and whole animal levels. It will result in state-of-the-art systems biology computational and experimental tools to enhance our understanding of cellular glycosylation, blood glycans, and its impact on human inflammatory diseases.