Detection and Histopathology Localization of O-Glycans and Glycosaminoglycans in Tissues

组织中 O-聚糖和糖胺聚糖的检测和组织病理学定位

• 批准号: 9166181
• 负责人: RICHARD R. DRAKE
• 金额: $ 27.36万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9166181
• 关键词: Address; Antibodies; Antigens; Binding; Biological; Biological Markers; Carbohydrates; Cataloging; Catalogs; Cell Extracts; Cell Line; Ceramides; Chondroitin; Chondroitin Sulfates; Chondroitinases; Detection; Digestion; Disaccharides; Disease; Drug or chemical Tissue Distribution; Family suidae; Formalin; Fourier transform ion cyclotron resonance; Freezing; Gangliosides; Glycosaminoglycans; Glycosphingolipids; Goals; Heparin; Heparin Lyase; Heparitin Sulfate; Histopathology; Image; Individual; Lectin; Link; Lipids; Malignant Neoplasms; Malignant neoplasm of pancreas; Maps; Mass Spectrum Analysis; Methods; Modification; Mucins; Mucopolysaccharidoses; Normal tissue morphology; Pathology; Physiological; Polysaccharides; Prostate; Proteins; Reaction; Reagent; Research; Research Personnel; Resolution; Resources; Role; Sialic Acids; Signal Transduction; Source; Spectrometry; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Staining method; Stains; Structure; Testing; Time; Tissue Extracts; Tissue Model; Tissue imaging; Tissues; abstracting; analytical method; analytical tool; aortic valve; cell type; comparative; imaging modality; improved; instrument; mouse model; novel; targeted treatment; tissue fixing; tissue preparation; two-dimensional

Abstract Extensive amounts of research has been done individually on the structure and functional roles ofthe four major glycan types (e.g., N-linked, O-linked, glycosaminoglycan (GAG) andglycosphingolipids (GSL)) in relation to disease states. How these different glycans are actuallydistributed within tissues has barely been studied, and frequently each class is studiedindependent of one another. Furthermore, analysis of these glycan classes are usually done intissue or cell extracts, forfeiting any localization opportunities. Any prior tissue mapping that hasbeen done has relied on broad class carbohydrate binding lectins, or carbohydrate antigenantibodies. These reagents may be used to localize structural glycans motifs within a giventissue, but they do not generally distinguish N-glycan or O-glycan proteins, nor GSL structures,as these could all share the same glycan structural target. To address this, our lab has recentlydeveloped a novel method to profile N-linked glycans directly on tissue using MALDI-imagingmass spectrometry using a high-resolution MALDI-FTICR instrument. This approach is mosteffective using formalin-fixed tissues, offering an unprecedented opportunity to analyze diseaseand normal formalin-fixed tissue blocks stored world-wide. The method is also very effective atidentification of both N-linked glycans and major GSL species in frozen tissues. The goal of thecurrent proposal is to develop related MALDI tissue imaging methods for O- linked mucin typeglycans and chondroitin/heparin GAG classes. This will provide unprecedented capabilities toexamine not only the individual distributions of these four glycan classes in tissues, but alsoallow comparisons of their distributions together in the same tissues. The proposal directlyaddresses the RFA-RM-15-008 goal of developing analytical methods and tools to enable rapidand detailed characterization of the complete glycan representation from a biological source.

抽象的 对结构和功能作用进行了大量研究 四种主要聚糖类型（例如，N-连接、O-连接、糖胺聚糖 (GAG) 和鞘糖脂（GSL））与疾病状态的关系。这些不同的聚糖有何不同 实际上在组织内的分布几乎没有被研究过，并且通常每一类都是 彼此独立地学习。此外，这些聚糖类别的分析通常是 完成组织或细胞提取物，失去任何本地化机会。任何先前的组织映射 已经完成的工作依赖于广泛的碳水化合物结合凝集素，或碳水化合物 抗原抗体。这些试剂可用于定位结构聚糖基序内 给定组织，但它们通常不区分 N-聚糖或 O-聚糖蛋白质，也不区分 GSL 结构，因为这些都可以共享相同的聚糖结构目标。为了解决这个问题，我们的实验室 最近开发了一种直接在组织上分析 N 连接聚糖的新方法，使用 使用高分辨率 MALDI-FTICR 仪器进行 MALDI 成像质谱分析。这 使用福尔马林固定组织的方法最为有效，提供了前所未有的机会 分析世界各地储存的疾病和正常福尔马林固定组织块。方法也是 非常有效地鉴定冷冻组织中的 N 连接聚糖和主要 GSL 种类。 当前提案的目标是开发O-相关的MALDI组织成像方法 连接粘蛋白型聚糖和软骨素/肝素 GAG 类。这将提供 前所未有的能力不仅可以检查这四种聚糖的个体分布 组织中的类别，而且还允许比较它们在相同组织中的分布。 该提案直接涉及 RFA-RM-15-008 开发分析方法和 能够快速、详细地表征完整聚糖表征的工具 生物来源。