Targeting Influenza A Virus by a Carbohydrate-inspired Strategy

通过碳水化合物启发的策略针对甲型流感病毒

• 批准号: 10458665
• 负责人: CHI-HUEY WONG
• 金额: $ 48.38万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-18 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10458665
• 关键词: Academia; Address; Aluminum Oxide; Antibodies; Antiviral Agents; Applications Grants; Binding; Biology; Capsid Proteins; Carbohydrates; Cell-Mediated Cytolysis; Cells; Cellular Membrane; Clinic; Collaborations; Collection; Complex; Detection; Development; Drug resistance; Engineering; Event; Genomics; Glass; Glycoengineering; Glycopeptides; Glycoproteins; HIV; HIV-1; Half-Life; Hemagglutinin; Immune; Immune system; Infection; Influenza; Influenza A virus; Influenza Therapeutic; Libraries; Life Cycle Stages; Malignant Neoplasms; Mediating; Membrane; Methods; Monoclonal Antibodies; Natural Killer Cells; Neuraminidase; Neuraminidase inhibitor; Oligosaccharides; Organism; Oseltamivir; Pathologic; Pharmaceutical Preparations; Play; Polysaccharides; Protein Biosynthesis; Protein Glycosylation; Proteins; Public Health; Research; Research Institute; Resistance; Respiratory System; Risk; Role; Sampling; Slide; Specificity; Structure; Structure-Activity Relationship; Techniques; Technology; Therapeutic; Therapeutic antibodies; Translating; Vaccines; Viral; Viral Hemagglutinins; Viral Proteins; Virus; anti-influenza; antibody-dependent cell cytotoxicity; base; cancer cell; clinical development; density; design; diagnostic tool; diagnostic value; drug resistant influenza; effective therapy; human pathogen; improved; in vivo; influenza infection; influenza virus strain; influenzavirus; intercellular communication; link protein; macrophage; neutralizing antibody; next generation; novel diagnostics; pandemic disease; receptor; receptor binding; recruit; sialic acid receptor; sialylation; zanamivir

PROJECT SUMMARY/ABSTRACT Protein glycosylation and carbohydrate-mediated recognition events play a major role during intercellular communications, which can engage components of the immune system and help maintain a healthy state of the organism. Unfortunately, many viruses can take advantage of protein glycosylation. In the case of influenza virus, the coat protein hemagglutinin (HA) mediates virus binding to sialoside receptors of the host cell and facilitates the fusion of viral and host membranes. Since complex-type N-glycosylation of proteins was shown to be crucial for the influenza virus infectivity, the putative influenza receptor is believed to be a glycoprotein. Unfortunately, N-glycan samples occupy only a small percentage of the established glycan collections that are used in the study of influenza virus. The availability of asymmetrically branched N-glycan samples is instrumental to the development of the next generation diagnostic tools and glycan sequencing methods. Using the modular method developed by our lab, we will expand the current collections of N-glycan structures. In order to improve diagnostic capabilities of glycan arrays for the profiling of influenza strains, we will create N-glycan arrays on the aluminium oxide coated glass (ACG) slides as this technology enables better control over the glycan density and distribution than conventional N-hydroxysuccinimide activated slides; and allows detection of even weak carbohydrate-based interactions. Synthetic samples of N-glycans will be used to optimize the anti-influenza broadly neutralizing antibody FI6, thus assisting clinical development of FI6 as universal and highly potent therapeutics of flu infection. The fragment crystallizable (Fc) region of FI6 contains a single N-glycan at N297, which engage FcγIIIa receptor, thus activating the antibody-depended cell-mediated cytotoxicity (ADCC) mechanism for neutralization and clearance of the infected cells. The current glycan structure-activity relationship (gSAR) is mostly established for the glycoforms with bi-antennary glycans, whereas protein glycoforms containing tetra-antennary complex-type N-glycans have not been accessed yet, even though, increasing the degree of sialylation is expected to improve ADCC and prolong in vivo half-life of glycoprotein therapeutics. We intend to complete the gSAR of FI6 by preparing glycoforms with tri- and tetra-antennary complex-type N-glycans, as well as neuraminidase-resistant N-glycan derivatives. New methods generated by this research and the expanded antibody gSAR will guide the general design of other therapeutic antibodies targeting cancer, HIV-1 and other pathological conditions, and will have a direct impact on public health.

项目概要/摘要 蛋白质糖基化和碳水化合物介导的识别事件在 细胞间通讯，可以参与免疫系统的组成部分并帮助维持 有机体的健康状态。不幸的是，许多病毒可以利用蛋白质糖基化。在 在流感病毒的情况下，外壳蛋白血凝素（HA）介导病毒与唾液酸苷受体结合 宿主细胞并促进病毒和宿主细胞膜的融合。由于蛋白质的复合型 N-糖基化 研究表明，流感病毒受体对流感病毒的传染性至关重要，因此假定的流感受体被认为是 糖蛋白。不幸的是，N-聚糖样品仅占已建立的聚糖的一小部分 用于流感病毒研究的集合。不对称支链 N-聚糖的可用性 样品有助于开发下一代诊断工具和聚糖测序 方法。使用我们实验室开发的模块化方法，我们将扩展当前的 N-聚糖系列 结构。为了提高聚糖阵列对流感毒株分析的诊断能力，我们 将在氧化铝涂层玻璃 (ACG) 载玻片上创建 N-聚糖阵列，因为该技术能够更好地实现 与传统的 N-羟基琥珀酰亚胺激活载玻片相比，对聚糖密度和分布的控制；和 甚至可以检测微弱的基于碳水化合物的相互作用。 N-聚糖的合成样品将用于优化抗流感广泛中和抗体 FI6，从而协助 FI6 作为通用且高效的流感感染疗法的临床开发。这 FI6 的片段可结晶 (Fc) 区域在 N297 处包含单个 N-聚糖，它与 FcγIIIa 受体结合， 从而激活抗体依赖性细胞介导的细胞毒性（ADCC）机制进行中和和 清除受感染的细胞。 目前的聚糖构效关系（gSAR）主要是针对以下糖型建立的： 双触角聚糖，而含有四触角复合型 N-聚糖的蛋白质糖型则没有 尽管如此，增加唾液酸化程度有望改善 ADCC 和 延长糖蛋白疗法的体内半衰期。我们打算通过准备来完成FI6的gSAR 具有三触角和四触角复合型 N-聚糖的糖型，以及耐神经氨酸酶的 N-聚糖 衍生物。本研究产生的新方法和扩展的抗体 gSAR 将指导一般 设计针对癌症、HIV-1 和其他病理状况的其他治疗抗体，并将 直接影响公众健康。