Glycophage arrays for the discovery of biomarkers in disease

用于发现疾病生物标志物的噬菌体阵列

• 批准号: 7611816
• 负责人: Adam Charles Fisher
• 金额: $ 20万
• 依托单位: GLYCOBIA, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2010-10-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7611816
• 关键词: Anabolism; Analytical Chemistry; Antigens; Asparagine; Bacteria; Bacteriophages; Biochemistry; Biological Markers; Biological Process; Blood; Campylobacter jejuni; Carbohydrates; Cell Adhesion; Cell Line; Cell surface; Cells; Cellular biology; Characteristics; Complex; Core Facility; Coupling; Custom; Deposition; Detection; Development; Diagnostic; Disease; Engineering; Ensure; Environment; Epitopes; Escherichia coli; Fluorescent Dyes; Foundations; Future; Galactosamine; Genetic; Glass; Glycoconjugates; Glycoproteins; Goals; HIV; Heart Diseases; Human; Immobilization; Learning; Life; Link; Lipids; Longevity; Lung; Malignant Neoplasms; Mammals; Methodology; Methods; Modeling; Organism; Pathology; Pattern; Phage Display; Play; Polysaccharides; Process; Production; Protein Glycosylation; Protein-Carbohydrate Interaction; Proteins; Reaction; Recombinants; Research; Retroviridae Infections; Robotics; Role; Scanning; Sensitivity and Specificity; Serum; Slide; Solid; Solutions; Source; Spottings; Staining method; Stains; Structure; Surface; System; Technology; Therapeutic; Therapeutic Agents; Universities; Variant; Work; analytical tool; base; cDNA Arrays; carbohydrate binding protein; carbohydrate structure; design; glycosylation; large scale production; link protein; method development; novel; particle; pathogen; public health relevance; research study; soybean lectin; structural biology; sugar; tool

DESCRIPTION (provided by applicant): Many biological processes throughout the lifespan of a living organism are governed by discrete sugar structures and their interaction with protein and lipid molecules. Such carbohydrates play essential roles in processes such as cell-cell recognition, adhesion, cell activation and host- pathogen interactions. In humans, variations in the synthesis of complex carbohydrates are known to cause pathologies including cancer, retrovirus infection and disorders of the heart, lung and blood. Substantial information has been learned about carbohydrate function from studies employing traditional analytical tools, yet there remains a demand for high-throughput methods that allow for the characterization and engineering of protein-carbohydrate interactions. High-throughput methods for characterizing carbohydrate/antigen interactions have been lacking for a variety of reasons including the absence of template driven synthesis and the significant structural complexity of glycans. However, even the current toolkit for detecting changes in glycan structure has shown great potential in the discovery of novel biomarkers for diseases such as cancer and HIV. Recently, it was discovered that the Campylobacter jejuni asparagine-linked (N-linked) protein glycosylation system can be functionally transferred into E. coli, conferring the ability to glycosylate proteins. Although the bacterial N-glycan is structurally different from its eukaryotic counterparts, such an accomplishment opens the door for engineered glycosylation reactions that mimic the processing of N-glycans in humans and other higher mammals. Glycobia seeks to apply such "bacterial glycoengineering" for the development of diagnostic and therapeutic agents. Towards this goal, the objective of this particular application is to synthesize a functional microarray of phage-displayed glycans (Aim 1) and use the microarray to detect basic protein-carbohydrate interactions (Aim 2). These studies should lay the necessary foundation for future work in expanding the diversity of phage-displayed glycans and analyzing carbohydrate signatures in complex serum. These studies are significant because they should (i) provide an analytical tool that overcomes the current bottlenecks of glycan microarray synthesis and (ii) enable the future synthesis of novel glycoconjugates for research, industrial and therapeutic applications. Public Health Relevance: Abnormalities in the assembly of complex sugars are known to be associated with a multitude of diseases including cancer, retrovirus infection and disorders of the heart, lung and blood. Carbohydrate-based array systems have emerged as the preferred methodology for analyzing carbohydrate-antigen interactions underlying these disorders, however the future utility of these arrays hinges critically on the ability to generate and recover diverse glycans for immobilization. The focus of these studies is the development of functional microarrays comprised of phage- displayed glycans for probing carbohydrate-antigen interactions.

描述（由申请人提供）：生物体整个生命周期中的许多生物过程都由离散的糖结构及其与蛋白质和脂质分子的相互作用控制。这些碳水化合物在细胞间识别、粘附、细胞激活和宿主-病原体相互作用等过程中发挥着重要作用。在人类中，复杂碳水化合物合成的变化已知会导致癌症、逆转录病毒感染以及心脏、肺和血液疾病等病症。通过使用传统分析工具的研究，人们已经了解了有关碳水化合物功能的大量信息，但仍然需要能够对蛋白质-碳水化合物相互作用进行表征和工程设计的高通量方法。由于多种原因，一直缺乏表征碳水化合物/抗原相互作用的高通量方法，包括缺乏模板驱动的合成以及聚糖的显着结构复杂性。然而，即使是目前用于检测聚糖结构变化的工具包，在发现癌症和艾滋病毒等疾病的新型生物标志物方面也显示出了巨大的潜力。最近，人们发现空肠弯曲杆菌天冬酰胺连接（N-连接）蛋白质糖基化系统可以功能性转移到大肠杆菌中，赋予蛋白质糖基化的能力。尽管细菌 N-聚糖在结构上与其真核对应物不同，但这一成就为工程化糖基化反应打开了大门，该反应可以模仿人类和其他高等哺乳动物中 N-聚糖的加工过程。 Glycobia 寻求应用这种“细菌糖工程”来开发诊断和治疗药物。为了实现这一目标，该特定应用的目标是合成噬菌体展示聚糖的功能微阵列（目标 1），并使用该微阵列检测基本的蛋白质-碳水化合物相互作用（目标 2）。这些研究应该为未来扩大噬菌体展示聚糖的多样性和分析复杂血清中碳水化合物特征的工作奠定必要的基础。这些研究意义重大，因为它们应该（i）提供一种分析工具，克服目前聚糖微阵列合成的瓶颈；（ii）使未来能够合成用于研究、工业和治疗应用的新型糖复合物。公共健康相关性：已知复合糖组装异常与多种疾病有关，包括癌症、逆转录病毒感染以及心脏、肺和血液疾病。基于碳水化合物的阵列系统已成为分析这些疾病背后的碳水化合物-抗原相互作用的首选方法，但是这些阵列的未来用途关键取决于生成和回收用于固定的多种聚糖的能力。这些研究的重点是开发由噬菌体展示的聚糖组成的功能性微阵列，用于探测碳水化合物-抗原相互作用。

项目成果

Production of secretory and extracellular N-linked glycoproteins in Escherichia coli.

大肠杆菌中分泌型和胞外 N 连接糖蛋白的生产。

• 发表时间: 2011-02
• 影响因子: 4.4
• 作者: Fisher, Adam C;Haitjema, Charles H;Guarino, Cassandra;Celik, Eda;Endicott, Christine E;Reading, Craig A;Merritt, Judith H;Ptak, A Celeste;Zhang, Sheng;DeLisa, Matthew P
• 通讯作者: DeLisa, Matthew P

Glycoarrays with engineered phages displaying structurally diverse oligosaccharides enable high-throughput detection of glycan-protein interactions.

具有显示结构多样化寡糖的工程化噬菌体的糖阵列能够高通量检测聚糖-蛋白质相互作用。

• 发表时间: 2015-01
• 影响因子: 4.7
• 作者: Çelik, Eda;Ollis, Anne A;Lasanajak, Yi;Fisher, Adam C;Gür, Göksu;Smith, David F;DeLisa, Matthew P
• 通讯作者: DeLisa, Matthew P