Novel HCV vaccine antigens and nanoparticles

新型 HCV 疫苗抗原和纳米颗粒

基本信息

批准号：
10557879
负责人：
Jiang Zhu
金额：
$ 56.97万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10557879
关键词：
Animal Model Antibodies Antibody Response Antibody-mediated protection Antigen Presentation Antigens Antiviral Agents Artificial nanoparticles B cell repertoire B-Lymphocytes Binding Biological Assay Cell Separation Cirrhosis Clinical Trials Complex Computer Models Data Developing Countries Development Engineering Epitopes Face Genetic Genotype Glycoproteins Goals Hepatitis C Vaccine Hepatitis C virus Heterodimerization Human Immune system Immunization Immunize Immunodominant Epitopes In Vitro Infection Knowledge Macaca mulatta Modeling Modification Molecular Conformation Mus Mutagenesis Outcome Polysaccharides Population Predisposition Primary carcinoma of the liver cells Production Property Proteins Public Health Reaction Recombinants Scaffolding Protein Scanning Site Structure Surface Transmembrane Domain Vaccination Vaccine Antigen Vaccine Design Vaccines Viral Viral Antigens Viral Envelope Proteins Virus clinical development combat cost effective design flexibility immunogenicity improved in vivo in vivo evaluation marginalization mouse model nanoparticle neutralizing antibody next generation sequencing nonhuman primate novel pandemic virus preference protein folding rational design response self assembly social group vaccine candidate vaccine development vaccine immunogenicity vaccine-induced antibodies virus envelope

Animal Model Antibodies Antibody Response Antibody-mediated protection Antigen Presentation Antigens Antiviral Agents Artificial nanoparticles B cell repertoire B-Lymphocytes Binding Biological Assay Cell Separation Cirrhosis Clinical Trials Complex Computer Models Data Developing Countries Development Engineering Epitopes Face Genetic Genotype Glycoproteins Goals Hepatitis C Vaccine Hepatitis C virus Heterodimerization Human Immune system Immunization Immunize Immunodominant Epitopes In Vitro Infection Knowledge Macaca mulatta Modeling Modification Molecular Conformation Mus Mutagenesis Outcome Polysaccharides Population Predisposition Primary carcinoma of the liver cells Production Property Proteins Public Health Reaction Recombinants Scaffolding Protein Scanning Site Structure Surface Transmembrane Domain Vaccination Vaccine Antigen Vaccine Design Vaccines Viral Viral Antigens Viral Envelope Proteins Virus clinical development combat cost effective design flexibility immunogenicity improved in vivo in vivo evaluation marginalization mouse model nanoparticle neutralizing antibody next generation sequencing nonhuman primate novel pandemic virus preference protein folding rational design response self assembly social group vaccine candidate vaccine development vaccine immunogenicity vaccine-induced antibodies virus envelope

展开

项目摘要

Project Summary Hepatitis C virus (HCV) infects 1-2% of the world population and poses a significant threat to public health. Recent studies have identified a panel of broadly neutralizing antibodies (bnAbs) and their genetic preferences. The crystal structures of various HCV E2 constructs in complex with bnAbs and non-nAbs provide a structural basis for rational vaccine design. In Project 2 of this P01 proposal, we will combine structural optimization of HCV antigens, nanoparticle engineering, in vivo assessment, and next-generation sequencing (NGS) of B-cell repertoires to assist in rational design of HCV vaccines that can elicit a bnAb response. The Specific Aims in this project are: (1) To develop vaccine antigens to target multidonor class antibody responses. The major virus neutralizing site, the E2 neutralizing face (E2 NF), is a well-known multidonor class antibody target on HCV. E2 NF is conformationally flexible and is surrounded and protected by immunodominant variable loops and N-glycans. The antigenic surface at and around E2 NF will be optimized by computational modeling and mutagenesis scanning to “lock” E2 NF into desired neutralizing conformations. A rigid E2 NF, together with modifications to improve protein folding and to minimize the decoy effect of immunodominant epitopes, should improve the immunogenicity of vaccine antigens and as a result elicit multidonor class bnAbs to the conserved neutralizing epitopes. (2) To develop E1E2-based vaccine antigens. In addition to E2 NF, the native E1E2 complex also present other conserved bnAb epitopes. We will apply scanning mutagenesis to the interface of E1 and E2 ectodomains and design novel E1 and E2 fusion constructs to improve the stability and production of the soluble E1E2 complex for the elicitation of bnAbs targeting the quaternary structure of the complex. (3) To develop self-assembling antigen-presenting nanoparticle vaccines. The engineered E2 and E1E2 complex will be fused with the scaffold proteins of different nanoparticle platforms to identify a platform optimal for the multimeric display of HCV antigens to the immune system. Based on our preliminary data, this strategy will greatly improve the immunogenicity of vaccine antigens and elicit a rapid and potent nAb response. (4) To evaluate immunogenicity and antibody responses of vaccine candidates in the mouse and non-human primate (NHP) rhesus macaque models. The engineered E2, E1E2 antigens and nanoparticles will first be studied in mice to confirm their immunogenicity in vivo and their ability to elicit antibodies that can bind and neutralize HCV. The antigens and nanoparticles with the best properties in vitro and in mice will be studied further using the NHP model. We have recently shown that rhesus macaques react to E1E2 immunization in a manner highly reminiscent to that used by humans and develop bnAbs against E2 NF with genetic similarity to human bnAbs. Here, we will immunize NHPs, identify bnAbs, and perform repertoire NGS to evaluate selected antigen designs. The outcome of Project 2 would be a set of well-characterized HCV vaccine candidates.

项目摘要丙型肝炎病毒（HCV）感染世界人口的1-2％，对公共卫生构成了重大威胁。最近的研究已经确定了一组广泛中和抗体（BNAB）及其遗传偏好。与BNAB和非NAB的复合物中各种HCV E2构建体的晶体结构提供了结构理性疫苗设计的基础。在本P01提案的项目2中，我们将结合结构优化 HCV抗原，纳米颗粒工程，体内评估和B细胞的下一代测序（NGS）曲目有助于合理设计可以引起BNAB响应的HCV疫苗。具体目的该项目是：（1）开发疫苗抗原以靶向多种抗体类抗体反应。这主要的病毒中和位点，E2中和面（E2 NF）是一个众所周知的多元抗体靶标在HCV上。 E2 NF在构象上是柔性的，并被免疫主导变量包围和保护循环和n-glycans。 E2 NF及其周围周围的抗原表面将通过计算建模进行优化并将诱变扫描以“锁定” e2 nf中和所需的中和构象。刚性E2 nf，一起修改以改善蛋白质折叠并最大程度地减少免疫主流表位的诱饵作用，应提高疫苗抗原的免疫原性，并因此将多配量bnabs带到配置中和表位。（2）开发基于E1E2的疫苗抗原。除了E2 NF，本机E1E2 复合物还提出其他保守的BNAB表位。我们将将扫描诱变应用于 E1和E2外生组和设计新颖的E1和E2融合构建体以提高稳定性和生产可溶性E1E2复合物用于启发靶向络合物第四纪结构的BNAB。（3）开发自组装抗原的纳米颗粒疫苗。工程的E2和E1E2 复合物将与不同纳米颗粒平台的脚手架蛋白融合，以识别最佳平台用于对免疫系统的HCV抗原的多媒体显示。基于我们的初步数据，此策略将大大改善疫苗抗原的免疫原性，并引起快速而潜在的NAB反应。（4）至评估小鼠和非人类疫苗候选疫苗的免疫原性和抗体反应灵长类动物（NHP）恒河猕猴模型。设计的E2，E1E2抗原和纳米颗粒将首先是在小鼠中研究以确认它们在体内的免疫原性，并引起可以结合和的抗体的能力中和HCV。在体外和小鼠中具有最佳特性的抗原和纳米颗粒将被研究进一步使用NHP模型。我们最近表明，恒河猕猴对A中的E1E2免疫反应高度让人联想到人类使用的方式，并针对E2 NF产生BNAB，与遗传相似人类bnabs。在这里，我们将免疫NHP，识别BNAB并执行曲目NGS来评估所选的抗原设计。项目2的结果将是一组特征良好的HCV疫苗候选物。