Automated chemo-enzymatic synthesis of N-glycans for host-pathogen interactions

用于宿主-病原体相互作用的 N-聚糖自动化学酶合成

• 批准号: 10626153
• 负责人: Geert-Jan Boons
• 金额: $ 42.48万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10626153
• 关键词: Address; Animals; Antibodies; Architecture; Automation; Avian Influenza A Virus; Binding; Binding Proteins; Biochemical Reaction; Biological; Biological Assay; Birds; Cell surface; Cells; Chemicals; Coin; Collection; Complex; Data; Dependence; Development; Disease Outbreaks; Engineering; Erythrocytes; Galactose; Glycoconjugates; Glycoproteins; Goals; Head; Hemagglutination; Hemagglutinin; Human; Immunization; Infection; Influenza A virus; Influenza vaccination; Intervention; Isomerism; Kinetics; Knowledge; Link; Measurement; Methodology; Modification; Mutation; Neuraminidase; Pathogenesis; Pattern; Phase; Polysaccharides; Preparation; Prevention strategy; Printing; Property; Protocols documentation; Reaction; Research; Respiratory System; Sialic Acids; Solid; Specificity; Speed; Structure; Sulfate; Surface; System; Time; Tissues; Transferase; Tropism; Vaccines; Viral; Virus; Virus Receptors; future pandemic; glycosylation; immunogenic; implementation facilitation; interest; next generation; novel; pathogen; pressure; programs; prototype; receptor; receptor binding; respiratory; respiratory virus; risk mitigation; seasonal influenza; sugar nucleotide; surveillance strategy; technology platform; trait; viral fitness

SUMMARY Numerous viruses initiate infection by binding to cell surface glycans of the host. The selectivities of viral receptor binding proteins for specific glycans critically determine host range, tissue- and cell tropism and pathogenesis. A detailed understanding of receptor usage by respiratory viruses is critical for the development of surveillance, prevention and intervention strategies to mitigate risks of future pandemic outbreaks. Glycan receptor usage by respiratory viruses have been difficult to probe because of a lack of appropriate panels of glycans for structure-activity studies. The latter is due to limitations in synthetic methodologies that do not permit the preparation of large panels of biological relevant glycans. In this program, chemoenzymatic methodologies will be developed that make it possible to prepare a wide range of N-glycan found in the respiratory tract of human and relevant animals. It is based on a new synthetic paradigm, which we coined “Stop-and-Go-Chemoenzymatic Glycosylation”. It uses chemically modified sugar nucleotide donors that can be employed by relevant glycosyl transferases to give products in which particular residues are temporarily blocked from further enzymatic modification. At an appropriate stage of synthesis, the blocking group can be removed to give a natural glycan. The speed of glycan synthesis will be increased by developing an automation platform that can perform enzymatic and chemical manipulations. The new methodologies will be used to prepare a collection of complex N-glycans that resemble structures expressed by respiratory tissue for host-pathogen studies. In this application, the collection of glycans will used to examine in detail receptor usage of influenza A viruses. The glycans will be printed as a microarray to probe binding specificities of human and animal influenza A viruses. Selected compounds will be examined in dynamic binding assays to establish the interplay between hemagglutinin (HA) and neuraminidase (NA) activity. The proposed studies will uncover unique traits of human and animal IAVs, which will facilitate the implementation of surveillance, prevention and intervention strategies to mitigate risks of future pandemics. The result of the studies will be exploited to develop an array-based system to antigenically characterize IAVs, which will greatly facilitate strain selection for seasonal flu vaccination. 1

概括 许多病毒通过与宿主的细胞表面糖质结合而引发感染。病毒的选择性 特异性聚糖的受体结合蛋白批判性确定的宿主范围，组织和细胞向量 和发病机理。对呼吸道病毒对受体使用的详细理解对于 制定监视，预防和干预策略以减轻未来大流行风险 爆发。由于缺乏，呼吸道病毒的聚糖受体使用很难探测 用于结构活性研究的合适的聚糖面板。后者是由于合成的限制 不允许制备大量生物学相关的聚糖的方法。 在该计划中，将开发化学酶方法学，使得准备 在人类和相关动物的呼吸道中发现了广泛的N-聚糖。它基于 我们创造了新的合成范式，我们创造了“停止和射酶糖基化”。它使用 化学修饰的糖核苷酸供体可以由相关的糖基转移到 给出特定残留物暂时阻止进一步酶促修饰的产品。 在合成的适当阶段，可以去除阻塞组以产生自然的聚糖。这 通过开发可以执行的自动化平台，将增加聚糖合成的速度 酶促和化学操作。新方法将用于准备 与呼吸道组织表达的结构进行宿主病原体研究相似的复杂N-聚糖。 在此应用中，聚集的集合将用于详细检查影响的受体用法 病毒。聚糖将被打印为探测人和动物的探测结合规格的微阵列 影响病毒。选定的化合物将在动态结合测定中检查以建立 血凝素（HA）和神经氨酸酶（Na）活性之间的相互作用。拟议的研究将 发现人类和动物IAV的独特特征，这将促进监视的实施 预防和干预策略来减轻未来大流行的风险。研究的结果将 探索以开发基于阵列的系统以抗原对IAV的特征，这将极大地 促进季节性流感疫苗接种的应变选择。 1