Methods for measuring matrisome molecule similarity during disease processes

测量疾病过程中基质体分子相似性的方法

• 批准号: 10330789
• 负责人: JOSEPH ZAIA
• 金额: $ 27.23万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10330789
• 关键词: Age; Aging; Animals; Award; Binding; Bioinformatics; Biological; Brain; Brain region; Cell Surface Proteins; Cells; Clinical Trials; Cytoplasmic Granules; Data; Detection; Disease; Extracellular Matrix; Focus Groups; Funding; Glycoproteins; Glycosaminoglycans; Goals; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Methods; Molecular; National Institute of General Medical Sciences; Neurodegenerative Disorders; Neurologic; Peptides; Periodicity; Physiological; Physiology; Polysaccharides; Process; Proteins; Proteoglycan; Proteomics; Research; Research Personnel; Sampling; Serine; Signal Transduction; Slide; Sulfate; Surface; Technology; Tissues; Water; analytical method; antibody detection; detection method; glycoprotein structure; glycoproteomics; glycosylation; instrument; ion mobility; liquid chromatography mass spectrometry; normal aging; pathogen; programs

Project Summary/Abstract: The application responds to NIGMS PAR-19-367 “Maximizing Investigators' Research Award (R35 - Clinical Trial Optional)” My research group focused initially on analytical methods for glycosaminoglycans (GAGs). These linear, sulfated polysaccharides are attached to serine residues of proteoglycan molecules found on cellular surfaces, in intracellular granules and in extracellular matrices. Binding interactions between GAGs and proteins are central to aspects of animal physiology including cellular signaling, the cellular microenvironment, and host- host, and host-pathogen recognition. My group developed methods for liquid chromatography-mass spectrometry analysis and sequencing of GAGs, extraction of GAGs from wet tissue and tissue slides, and bioinformatics programs for interpretation of GAG MS and tandem MS data. We then pioneered methods to acquire glycomics (GAGs and N-glycans) and proteomics from biospecimen tissue slides. We applied these methods to study of neurological aging, cancer, neurodevelopmental, and neurodegenerative diseases. As funded through NIGMS R01GM133963 “Methods for determination of glycoprotein glycosylation similarities among disease states”, my group is developing analytical and bioinformatics methods for glycoproteomics of the extracellular matrix molecules (known as the matrisome) of brain. The primary focus has been on glycoproteomics MS acquisition methods and bioinformatics for rigorous statistical determination of molecular similarities for matrisome molecules and how these change during normal aging versus disease processes. I now propose to expand our brain glycoproteomics molecular similarity scope to include characterization of GAGs that modify specific matrisome molecules. Our goals will be to provide information on the pathophysiological changes to matrisome molecules that escape detection using traditional antibody-based detection methods. We will employ a new Omnitrap platform to characterize multiply glycosylated peptides and peptides modified with GAG chains. We will also exploit the capabilities of ion mobility for measuring molecular similarities from glycoproteomics data using a Waters Cyclic ion mobility-mass spectrometry instrument. We will demonstrate these approaches by comparing matrisome molecular similarity among brain regions and as a function of age.

项目摘要/摘要： 该应用程序响应 NIGMS PAR-19-367“最大化研究者研究奖（R35 - 临床 试用可选）” 我的研究小组最初专注于糖胺聚糖 (GAG) 的分析方法。 硫酸化多糖附着在细胞表面的蛋白聚糖分子的丝氨酸残基上， 在细胞内颗粒和细胞外基质中，GAG 和蛋白质之间存在结合相互作用。 动物生理学的核心，包括细胞信号传导、细胞微环境和宿主 我的小组开发了液相色谱-质谱法的方法。 GAG 的光谱分析和测序，从湿组织和组织玻片中提取 GAG，以及 然后我们开创了解释 GAG MS 和串联 MS 数据的生物信息学程序。 我们应用这些从生物样本组织玻片中获取糖组学（GAG 和 N-聚糖）和蛋白质组学。 研究神经衰老、癌症、神经发育和神经退行性疾病的方法。 由 NIGMS R01GM133963“糖蛋白糖基化相似性测定方法”资助 在疾病状态中”，我的小组正在开发糖蛋白组学的分析和生物信息学方法 主要关注点是大脑的细胞外基质分子（称为基质体）。 糖蛋白组学 MS 采集方法和生物信息学，用于分子的严格统计测定 基质体分子的相似性以及它们在正常衰老与疾病过程中如何变化。 我现在建议扩大我们的大脑糖蛋白组分子相似性范围，以包括以下特征： 我们的目标是提供有关特定基质体分子的 GAG。 基质体分子的病理生理变化逃避了传统基于抗体的检测 我们将采用新的 Omnitrap 平台来表征多重糖基化肽和 我们还将利用离子淌度测量分子的能力。 使用沃特世循环离子淌度质谱仪获得的糖蛋白组学数据的相似性。 将通过比较大脑区域之间的基质体分子相似性并作为 年龄的函数。