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.

项目摘要/摘要 蛋白质糖基化和碳水化合物介导的识别事件在 细胞间通信，可以参与免疫系统的组件并帮助维护 健康状态。不幸的是，许多病毒可以利用蛋白质糖基化。在 造影症病毒的病例，外套蛋白血凝素（HA）介导了与结合的病毒与唾液酸受体的结合 宿主细胞并促进病毒和宿主机制的融合。由于蛋白质的复合型N-糖基化 被证明对于影响病毒感染至关重要，假定的影响者被认为是 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 在影响扎病毒研究中使用的收集。非对称分支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。使用实验室开发的模块化方法，我们将扩展N-Glycan的当前集合 结构。为了提高聚糖阵列的诊断能力以进行影响菌株的分析，我们 将在此技术上更好地在氧化铝涂层玻璃（ACG）滑梯上创建N-聚糖阵列 控制聚糖密度和分布比常规的N-羟基糖二酰亚胺激活的载玻片的控制；和 允许检测甚至弱碳水化的相互作用。 N-聚糖的合成样品将用于优化抗Influenza广泛中和抗体 FI6，因此有助于FI6作为流感感染的普遍且高潜在的治疗。 FI6的碎片可结晶（FC）区域在N297上包含一个N-Glycan，它与FcγIIIA受体接合， 因此激活抗体依赖性细胞介导的细胞毒性（ADCC）神经化和 被感染细胞的清除。 当前的聚糖结构 - 活性关系（GSAR）主要是针对与糖型建立的 双静脉聚糖，而含有四抗复合物型N-聚糖的蛋白质糖型尚未 即使可以提高溶解酶的程度，也可以提高ADCC和 延长糖蛋白疗法的体内半衰期。我们打算通过准备FI6的GSAR 具有三 - 抗复合物型N-聚糖的糖型，以及耐神经苷酶的N-聚糖 衍生物。这项研究产生的新方法和扩展的抗体GSAR将指导一般 针对癌症，HIV-1和其他病理状况的其他治疗抗体的设计，并将具有 对公共卫生的直接影响。