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: 9320983
• 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 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9320983
• 关键词: Address; Antibodies; Antigens; Binding; Biological; Biological Markers; Carbohydrates; 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; GAG Gene; 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; analytical method; analytical tool; aortic valve; cell type; chemical release; comparative; imaging modality; improved; instrument; mouse model; novel; targeted treatment; 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 大量研究已在结构和功能角色上单独进行 在四种主要聚糖类型（例如，N连接，O连接，糖胺聚糖（GAG）） 与疾病状态有关的糖磷脂（GSL）。这些不同的聚糖如何 实际上几乎没有研究在组织内部的分布，每个班级经常是 研究彼此独立。此外，对这些聚糖类的分析通常是 完成了静电或细胞提取物，没收任何本地化机会。任何先前的组织映射 Hosbeen完成的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。这些试剂可用于将结构聚糖基序定位在 Giventissue，但通常不会区分N-聚糖或O-聚糖蛋白，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 最近开发了一种新的方法，可以直接在组织上直接在组织上介绍N连接的聚糖 使用高分辨率MALDI-FEFTICR仪器的MALDIMIMAGGANGASS光谱法。这 使用福尔马林固定的组织最有效的方法，提供了前所未有的机会 分析福尔马林固定的组织块疾病全球存储的疾病。该方法也是 在冷冻组织中非常有效地识别N连锁的聚糖和主要GSL物种。 提议的目的是开发相关的MALDI组织成像方法 连接的粘蛋白类型糖和软骨素/肝素插入类。这将提供 前所未有的功能，不仅是这四个聚糖的单个分布 组织中的类别，但还可以在同一组织中一起对其分布进行比较。 该提案直接添加了RFA-RM-15-008开发分析方法和 实现快速和详细表征完整的聚糖表示形式的工具 生物来源。