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的，化学上的复杂且生物学上不同的翻译后 涉及各种细胞功能的修改（PTM）。糖基化在调节中起着至关重要的作用 细胞增殖和代谢过程以及糖基化的变化是普遍特征 恶性转化和肿瘤进展。转移，或癌症传播到非粘量的部分 身体是许多侵略性癌症的特别阴险的特征。证据表明特定 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攻击/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，但当前的技术用于表征 该PTM不发达。质谱法（MS）是用于分析PTM的黄金标准，但是化学 相对于其他修饰，糖基化的复杂性显着放缓了MS技术的进度。 该提案引入了激活的离子电子传递解离（AI-ETD）作为新的串联MS方法 为了全面表征完整的糖肽。红外照片激活和 AI-ETD中电子驱动的自由基碎片产生肽序列和聚糖组成信息 在单个串联MS事件中。这消除了每个先前的多个串联MS扫描，这是必要的 当前的方法，提高灵敏度并有效增加吞吐量。在此应用程序中，AI-ETD将是 在最新一代的Orbitrap MS系统（Fusion Lumos）上实施，以利用其强大的数据 - 将开发获取平台，以及用于糖蛋白质组学实验的高通量AI-ETD方法。 这些方法将用于三种等源性人类癌细胞系的糖蛋白质组学表征， 表示黑色素瘤的非，中间和高度转移性形式。通过启用表征 数百种糖材料和数千种聚糖，这项工作将是最全面的糖蛋白质组学 癌细胞系的比较，允许研究癌症攻击的糖基化特征 整个糖蛋白酶的空前的宽度和深度。该数据集将提高对 分子水平的癌症转移，并揭示了糖基化在癌症中的作用的新见解。 该项目完成后，我将寻求杰出癌症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。结合 MS仪器开发和癌症的聚糖方面的专业知识将为我提供多产 在生物学和技术界面以及癌症研究的最前沿的独立科学家职业。

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.