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

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.

项目概要丙型肝炎病毒 (HCV) 感染世界人口的 1-2%，对公众健康构成重大威胁。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 复合的各种 HCV E2 构建体的晶体结构提供了结构在这个P01提案的项目2中，我们将结合结构优化来进行合理的疫苗设计。 HCV 抗原、纳米颗粒工程、体内评估和 B 细胞下一代测序 (NGS) 有助于合理设计能够引发 bnAb 反应的 HCV 疫苗。该项目是： (1) 开发针对多供体类抗体反应的疫苗抗原。主要病毒中和位点，E2 中和面 (E2 NF)，是众所周知的多供体类抗体靶点 HCV 上的 E2 NF 构象灵活，并受到免疫显性变量的包围和保护。 E2 NF 及其周围的抗原表面将通过计算建模进行优化。诱变扫描将 E2 NF 与刚性 E2 NF 一起“锁定”为所需的中和构象。修饰以改善蛋白质折叠并尽量减少免疫显性表位的诱饵效应，应该提高疫苗抗原的免疫原性，从而引发保守的多供体类 bnAb (2) 开发基于E1E2的疫苗抗原，除了E2 NF外，还有天然的E1E2。复合物还呈现其他保守的bnAb表位，我们将对其界面应用扫描诱变。 E1 和 E2 胞外域并设计新型 E1 和 E2 融合构建体以提高稳定性和产量可溶性 E1E2 复合物，用于引发针对该复合物四级结构的 bnAb。开发自组装抗原呈递纳米颗粒疫苗。复合物将与不同纳米颗粒平台的支架蛋白融合，以确定最佳平台根据我们的初步数据，该策略用于向免疫系统展示 HCV 抗原的多聚体。 (4) 至评估候选疫苗在小鼠和非人类体内的免疫原性和抗体反应工程化的 E2、E1E2 抗原和纳米颗粒将首先用于灵长类动物 (NHP) 恒河猴模型。在小鼠中进行研究，以确认它们的体内免疫原性以及它们引发可以结合和的抗体的能力将研究在体外和小鼠体内具有最佳特性的抗原和纳米颗粒。我们最近进一步使用 NHP 模型表明，恒河猴对 E1E2 免疫有反应。以与人类使用的方式高度隐蔽的方式开发针对 E2 NF 的 bnAb，其遗传相似性在这里，我们将对 NHP 进行免疫，鉴定 bnAb，并进行全谱 NGS 来评估所选的项目 2 的成果将是一组特征明确的 HCV 候选疫苗。