Uniting Mass Spectrometry and Glycoscience to Investigate Cancer Biology

结合质谱和糖科学来研究癌症生物学

• 批准号: 10240582
• 负责人: Nicholas M Riley
• 金额: $ 8.77万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-13 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10240582
• 关键词: Aggressive behavior; Amino Acid Sequence; Biochemical; Biochemistry; Biodiversity; Biological; Biology; Body part; Cancer Biology; Cancer cell line; Cell Line; Cell Proliferation; Cell physiology; Cellular biology; Characteristics; Chemicals; Chemistry; Communities; Complement; Complex; Coupled; Data Set; Development; Dissociation; Electron Transport; Electrons; Environment; Event; Fostering; Foundations; Generations; Glycobiology; Glycopeptides; Goals; Gold; Heterogeneity; Human; Hybrids; Interdisciplinary Study; Investigation; Ions; Knowledge; Laboratories; Length; Link; Malignant - descriptor; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Metabolism; Methodology; Methods; Modification; Molecular; Molecular Biology; Neoplasm Metastasis; Nonmetastatic; Oncogenic; Peptides; Phase; Play; Polysaccharides; Post Translational Modification Analysis; Post-Translational Modification Site; Post-Translational Protein Processing; Postdoctoral Fellow; Property; Protein Analysis; Protein Glycosylation; Protein Region; Proteins; Proteome; Proteomics; Reaction; Research; Research Personnel; Resolution; Role; Scanning; Scientist; Site; Speed; Structure; System; Technical Expertise; Techniques; Technology; Training; Work; analytical method; anticancer research; cancer cell; career; cell growth regulation; data acquisition; experimental study; fitness; frontier; glycoproteomics; glycosylation; improved; innovation; insight; instrument; instrumentation; melanoma; photoactivation; post-doctoral training; pre-doctoral; programs; protein aminoacid sequence; specific biomarkers; success; tandem mass spectrometry; technology development; tumor progression

Project Summary/Abstract. Protein glycosylation is a prevalent, chemically complex, and biologically diverse post-translational modification (PTM) involved in a wide array of cellular functions. Glycosylation plays essential roles in regulation of cellular proliferation and metabolic processes, and changes in glycosylation are universal features of malignant transformation and tumor progression. Metastasis, or the spread of cancer to non-adjacent parts of the body, is a particularly insidious characteristic of many aggressive cancers. Evidence suggests that specific changes in glycosylation may reflect fitness of tumor progression and metastatic potential, making global characterization of glycosylation crucial to understanding the molecular basis of cancer aggression/metastasis. Despite the critical importance of glycosylation in cancer research, current technology for characterizing this PTM is underdeveloped. Mass spectrometry (MS) is the gold standard for analysis of PTMs, but the chemical complexity of glycosylation has significantly slowed progress of MS technology relative to other modifications. This proposal introduces activated-ion electron transfer dissociation (AI-ETD) as a new tandem MS approach for comprehensive characterization of intact glycopeptides. The combination of infrared photo-activation and electron-driven radical fragmentation in AI-ETD generates peptide sequence and glycan composition information in a single tandem MS event. This eliminates multiple tandem MS scans per precursor that are necessary in current approaches, improving sensitivity and effectively doubling throughput. In this application, AI-ETD will be implemented on the newest generation of Orbitrap MS systems (Fusion Lumos) to capitalize on its robust data- acquisition platform, and high-throughput AI-ETD methods for glycoproteomic experiments will be developed. These methods will be utilized in glycoproteomic characterization of three isogenic human cancer cell lines that represent non-, intermediate-, and highly-metastatic forms of melanoma. By enabling characterization of hundreds of glycosites and thousands of glycans, this work will be the most comprehensive glycoproteomic comparison of cancer cell lines yet, allowing investigation of glycosylation signatures of cancer aggression with unprecedented breadth and depth across the glycoproteome. This data set will advance the understanding of cancer metastasis at a molecular level and reveal new insights into the role of glycosylation in cancer. Upon completion of this project, I will seek postdoctoral training in a preeminent cancer glycobiology laboratory, complementing the technology development focus of my graduate research. The combination of expertise in MS instrumentation development and the glycoscience of cancer will uniquely equip me for a prolific career as an independent scientist at the interface of biology and technology and the forefront of cancer research.

项目摘要/摘要。 蛋白质糖基化是一种普遍存在的、化学性质复杂且生物多样性的翻译后修饰 修饰（PTM）涉及广泛的细胞功能。糖基化在调节中发挥重要作用 细胞增殖和代谢过程的变化以及糖基化的变化是细胞增殖和代谢过程的普遍特征 恶性转化和肿瘤进展。转移，或癌症扩散到非邻近部位 身体，是许多侵袭性癌症的一个特别阴险的特征。有证据表明，具体 糖基化的变化可能反映肿瘤进展和转移潜力的适应性，使全球 糖基化的表征对于理解癌症侵袭/转移的分子基础至关重要。 尽管糖基化在癌症研究中至关重要，但目前的表征技术 该 PTM 尚未开发。质谱 (MS) 是分析 PTM 的黄金标准，但化学分析 相对于其他修饰，糖基化的复杂性显着减缓了 MS 技术的进展。 该提案引入了活化离子电子转移解离 (AI-ETD) 作为一种新的串联 MS 方法 用于完整糖肽的综合表征。红外光活化和 AI-ETD 中的电子驱动自由基碎裂生成肽序列和聚糖组成信息 在单个串联 MS 事件中。这消除了每个前体所需的多次串联 MS 扫描 当前的方法，提高灵敏度并有效地使吞吐量加倍。在此应用中，AI-ETD 将 在最新一代 Orbitrap MS 系统 (Fusion Lumos) 上实施，以利用其强大的数据- 将开发用于糖蛋白质组实验的高通量AI-ETD方法。 这些方法将用于三种同基因人类癌细胞系的糖蛋白组学表征， 代表非、中间和高度转移形式的黑色素瘤。通过启用表征 数百个糖位点和数千个聚糖，这项工作将是最全面的糖蛋白组学 尚未对癌细胞系进行比较，从而可以研究癌症侵袭的糖基化特征 糖蛋白组前所未有的广度和深度。该数据集将加深对 在分子水平上研究癌症转移，并揭示糖基化在癌症中的作用的新见解。 完成该项目后，我将寻求卓越的癌症糖生物学的博士后培训 实验室，补充了我研究生研究的技术开发重点。的组合 MS 仪器开发和癌症糖科学方面的专业知识将为我提供独特的多产能力 作为一名独立科学家，从事生物学和技术领域以及癌症研究前沿的工作。

项目成果

MYC-driven synthesis of Siglec ligands is a glycoimmune checkpoint.

• DOI: 10.1073/pnas.2215376120
• 发表时间: 2023-03-14
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Smith, Benjamin A. H.;Deutzmann, Anja;Correa, Kristina M.;Delaveris, Corleone S.;Dhanasekaran, Renumathy;Dove, Christopher G.;Sullivan, Delaney K.;Wisnovsky, Simon;Stark, Jessica C.;V. Pluvinage, John;Swaminathan, Srividya;Riley, Nicholas M.;Rajan, Anand;Majeti, Ravindra;Felsher, Dean W.;Bertozzi, Carolyn R.
• 通讯作者: Bertozzi, Carolyn R.

Lysosomal cathepsin D mediates endogenous mucin glycodomain catabolism in mammals.

• DOI: 10.1073/pnas.2117105119
• 发表时间: 2022-09-27
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1

Synthetic Siglec-9 Agonists Inhibit Neutrophil Activation Associated with COVID-19.

合成 Siglec-9 激动剂抑制与 COVID-19 相关的中性粒细胞激活。

• DOI: 10.26434/chemrxiv.13378148
• 发表时间: 2020
• 期刊: ChemRxiv : the preprint server for chemistry
• 作者: Delaveris,CorleoneS;Wilk,AaronJ;Riley,NicholasM;Stark,JessicaC;Yang,SamuelS;Rogers,AngelaJ;Ranganath,Thanmayi;Nadeau,KariC;StanfordCOVID-19Biobank;Blish,CatherineA;Bertozzi,CarolynR
• 通讯作者: Bertozzi,CarolynR

Targeting hypersialylation in multiple myeloma represents a novel approach to enhance NK cell-mediated tumor responses.

• DOI: 10.1182/bloodadvances.2021006805
• 发表时间: 2022-06-14
• 期刊: BLOOD ADVANCES
• 影响因子: 7.5
• 作者: Daly, John;Sarkar, Subhashis;Natoni, Alessandro;Stark, Jessica C.;Riley, Nicholas M.;Bertozzi, Carolyn R.;Carlsten, Mattias;O'Dwyer, Michael E.
• 通讯作者: O'Dwyer, Michael E.