Structural analysis of HCV E1E2 glycoproteins

HCV E1E2 糖蛋白的结构分析

• 批准号: 10205552
• 负责人: Richard J. Kuhn
• 金额: $ 31.24万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10205552
• 关键词: Animals; Antibodies; Antigens; Architecture; B-Lymphocytes; Binding; Bioinformatics; Biology; Biophysics; CD81 gene; Cavia; Complex; Computer Models; Cryo-electron tomography; Cryoelectron Microscopy; Crystallography; Data; Electron Microscopy Facility; Engineering; Epitopes; Fc Receptor; Flavivirus; Genotype; Glycoproteins; Goals; Hepatitis C; Hepatitis C Vaccine; Hepatitis C virus; Human; Immunity; Immunology procedure; Laboratories; Lead; Length; Light; Mediating; Membrane Glycoproteins; Methods; Modification; Molecular Conformation; Molecular Virology; Mutation; Nature; Outcome; Polysaccharides; Preparation; Production; Protein Engineering; Proteins; Protocols documentation; Recombinant Proteins; Recombinants; Research; Resolution; Role; Structure; Surface; Synchrotrons; T-Lymphocyte; Testing; Time; Transmembrane Domain; Vaccine Design; Vaccines; Viral; Viral Pathogenesis; Virion; Virus; Virus Receptors; Work; X-Ray Crystallography; base; design; experience; glycoprotein structure; glycosylation; human pathogen; in vivo; insight; lead candidate; molecular modeling; nanodisk; neutralizing antibody; nonhuman primate; novel; particle; plasmid DNA; programs; receptor; receptor binding; response; structural biology; tool

ABSTRACT – PROJECT 4 Despite extensive research efforts to understand the biology of hepatitis C virus (HCV), it remains a major human pathogen without a vaccine. In addition, there is a remarkable absence of information regarding the structure of the virion and organization of its component proteins. We intend to use our extensive structural biology experience with the related flaviviruses, to focus on determining the structure of the viral glycoproteins, the virus particle, and complexes of potent neutralizing antibodies bound to the surface glycoproteins. This project will rely heavily on and interact extensively with Project 1 to interrogate the structure and provide guidance into the features of the glycoproteins that are important for eliciting broad and potent neutralizing antibodies. Iterative analysis and re-engineering of the E1E2 glycoproteins will be conducted to guide the antibody component of a dual B and T cell HCV vaccine, the focus of this U19 proposal. This specific project will use three aims to determine the structure of the E1E2 heterodimer, the virus particle, and neutralizing antibody Fabs bound to the glycoproteins. Based on biophysical and immunological assays, computational modeling and structure analyses, the E1E2 glycoproteins will be engineered to present epitopes that elicit strong neutralizing and broadly reactive antibodies and minimize the presentation of decoy non-productive epitopes. Furthermore, the organization of the glycoproteins and the structure of the virus particle will be determined. The tools of molecular virology, biophysics, computational modeling, x-ray crystallography and cryo-electron microscopy will be used to produce atomic-level details of the epitope landscape of HCV. The gained structural information will be utilized to design novel antigens in Project 1, determine the structures of designed immunogens experimentally in Project 4 and tested in vivo in Projects 1, 2 and 3. We will guide Project 1 at various times during the project to allow them to utilize the optimal E1E2 immunogen for studies in guinea pigs and, ultimately, a lead candidate for an optimized E1E2 immunogen for nonhuman primate studies. This information will be invaluable in deciphering the nature of neutralizing antibodies and their mechanisms of action against HCV and will shed light on the assembly and entry of hepatitis C virions.

摘要 - 项目4 尽管广泛的研究努力了解丙型肝炎病毒（HCV）的生物学，但它仍然是主要的人类 没有疫苗的病原体。此外，关于结构的明显缺乏 其成分蛋白的病毒和组织。我们打算使用广泛的结构生物学 具有相关黄病毒的经验，专注于确定病毒糖蛋白的结构，病毒 与表面糖蛋白结合的潜在中和抗体的颗粒和复合物。这个项目将依靠 大量与项目1进行广泛互动，以询问结构并为 糖蛋白的特征对于引起广泛而潜在的中和抗体很重要。迭代 将进行E1E2糖蛋白的分析和重新设计，以指导A的抗体成分 双B和T细胞HCV疫苗，该U19提案的重点。这个特定项目将使用三个目标 确定E1E2异二聚体，病毒颗粒的结构，并中和抗体晶圆厂结合 糖蛋白。基于生物物理和免疫学测定，计算建模和结构分析， E1E2糖蛋白将被设计为提出引起强烈中和和广泛反应性的表位 抗体并最大程度地减少诱饵非生产表位的表现。此外，组织 将确定糖蛋白和病毒颗粒的结构。分子病毒学工具， 生物物理学，计算建模，X射线晶体学和冷冻电子显微镜将用于产生 HCV表位景观的原子级细节。获得的结构信息将用于设计 项目1中的新型抗原，在项目4和 在项目1、2和3的体内测试。我们将在项目期间的不同时间指导项目1，以允许他们 利用最佳E1E2免疫原进行豚鼠的研究，并最终成为优化的主要候选者 非人类灵长类动物研究的E1E2免疫原。这些信息在解密的性质中是无价的 中和抗体及其针对HCV的作用机制，并将阐明组件和 肝炎病毒的进入。