Design and evaluation of pan-CoV vaccines

泛冠状病毒疫苗的设计和评估

• 批准号: 10841733
• 负责人: YOSHIHIRO KAWAOKA
• 金额: $ 281.95万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-16 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10841733
• 关键词: 2019-nCoV; Amino Acids; Animal Model; Animals; Antibodies; Antibody Response; Antibody titer measurement; Antigens; B-Lymphocytes; Chicago; Chimera organism; Collaborations; Complex; Coronavirus; Coronavirus spike protein; Cryoelectron Microscopy; Data; Development; Encapsulated; Enterobacteria phage MS2; Epitopes; Evaluation; Funding Opportunities; Goals; Head; Hearing; Human; Immune; Immune response; Immune system; Immunization; Immunize; Immunodominant Epitopes; Immunology; Individual; Influenza Hemagglutinin; Japan; Laboratories; Mesocricetus auratus; Messenger RNA; Modification; Molecular Virology; Mus; Mutation; Pharmacologic Substance; Phase I Clinical Trials; Polysaccharides; Positioning Attribute; Pre-Clinical Model; Proteins; Research Project Grants; SARS-CoV-2 spike protein; Saint Jude Children' s Research Hospital; Sampling; Structural Biologist; System; T cell response; T-Lymphocyte; Testing; Universities; Vaccinated; Vaccination; Vaccines; Vesicular stomatitis Indiana virus; Viral Antigens; Virus; Virus-like particle; Wisconsin; X-Ray Crystallography; candidate selection; comparison control; coronavirus vaccine; cross reactivity; design; flexibility; immunogenicity; influenza virus vaccine; influenzavirus; lipid nanoparticle; mutant; nanobiotechnology; novel; novel coronavirus; progenitor; protective efficacy; reconstruction; self assembly; stem; universal coronavirus vaccine; vaccine candidate; vaccine development; vaccine platform; virology

SUMMARY As part of the Pan-Coronavirus Vaccine (PanCoVac) Consortium, the goal of Research Project 1 (RP1) is the ‘Design and evaluation of pan-CoV vaccines’. A comprehensive characterization of the B and T cell responses to vaccination will be carried out by Research Project 2 (RP2, ‘Immunological responses to pan-CoV vaccines. In Aim 1 of RP1 (‘Focusing immune responses towards the stem of the spike protein’), several strategies will be tested to increase the levels of antibodies directed at the conserved, immune-subdominant epitopes in the stem of the SARS-CoV-2 spike protein (to major viral antigen), while avoiding strong antibody responses directed at immunodominant epitopes in the head of the spike protein. These strategies include chimeric spikes composed of SARS-CoV-2 immune-subdominant stem epitopes and immunodominant head epitopes of unencountered coronaviruses, ‘outdiluting’ antibody responses to the immunodominant epitopes by using cocktails of spike proteins with multiple mutations in key amino acid positions, glycan-shielding of immunodominant head epitopes, ‘inverted antigens’ in which the spike protein will be presented to the immune system in an inverted orientation (and thus become more accessible), and ‘headless’ spike proteins lacking portions of the immunodominant head epitopes. Some of these approaches may be tested in combination, and may be tested with sequence-optimized stem epitopes based on ancestral reconstruction (a computational approach to deduce the most likely common progenitor sequence). In Aim 2 (‘Eliciting broadly reactive immune responses to the head of the spike protein’), strategies will be tested to direct the immune responses away from the most sequence-diverse epitopes and towards more conserved epitopes in the head. In addition, conserved heard epitopes will be sequenced-optimized to be recognized by cross-protective antibodies. These modifications will be introduced into diverse spike proteins, and vaccine cocktails of diverse, mutant spike proteins will then be tested for their immunogenicity and protective efficacy. All antigens in Aims 1 and 2 will be designed in collaboration with a structural biologist, and for selected antigens and/or antigen/antibody complexes, X-ray crystallography and Cryo-EM will be carried out. In Aim 3 (‘Immunogenicity and protective efficacy of broadly reactive antigens’), the novel vaccinates will be tested in mice for their immunogenicity; samples from vaccinated mice will be provided to RP2 for B and T cell analysis. Selected vaccine candidates (those with broader immune responses) will next be tested in mice and Syrian hamsters for their ability to provide protection against different coronaviruses. For candidate vaccines that provide broad protection (compared to controls), we will also assess the durability of immune responses, and the effect of vaccination on virus transmissibility. Moreover, these vaccinate candidates will be tested in an mRNA lipid nanoparticle vaccine platform provided by Daiichi Sankyo, a pharmaceutical company.

概括 作为泛冠状病毒疫苗 (PanCoVac) 联盟的一部分，研究项目 1 (RP1) 的目标是 “泛冠状病毒疫苗的设计和评估”。 B 细胞和 T 细胞的综合表征 对疫苗接种的反应将由研究项目 2（RP2，“对泛冠状病毒的免疫反应”）进行 疫苗。在 RP1 的目标 1（“将免疫反应集中于刺突蛋白的茎”）中，几个 将测试策略以增加针对保守的、免疫次优势的抗体的水平 SARS-CoV-2 刺突蛋白茎中的表位（针对主要病毒抗原），同时避免强抗体 针对刺突蛋白头部的免疫显性表位的反应。这些策略包括 由 SARS-CoV-2 免疫亚显性茎表位和免疫显性头组成的嵌合刺突 未遇到的冠状病毒的表位，通过“稀释”抗体对免疫显性表位的反应 使用在关键氨基酸位置具有多个突变的刺突蛋白混合物，聚糖屏蔽 免疫显性头部表位，“反向抗原”，其中刺突蛋白将呈递给免疫系统 系统处于倒置方向（因此变得更容易接近），并且缺乏“无头”刺突蛋白 免疫显性头部表位的部分。其中一些方法可以组合进行测试，并且 可以使用基于祖先重建（计算 推导最可能的共同祖先序列的方法）。目标 2（“引发广泛的反应 对刺突蛋白头部的免疫反应”），将测试指导免疫反应的策略 远离序列最多样化的表位并朝向头部中更保守的表位。此外， 保守的表位将被测序优化以被交叉保护性抗体识别。这些 修饰将被引入不同的刺突蛋白以及不同突变刺突的疫苗混合物中 然后将测试蛋白质的免疫原性和保护功效。目标 1 和 2 中的所有抗原都将 与结构生物学家合作设计，针对选定的抗原和/或抗原/抗体复合物， 将进行 X 射线晶体学和冷冻电镜分析。目标 3（“免疫原性和保护功效”） 广泛反应性抗原”），新型疫苗将在小鼠中测试其免疫原性；样本来自 接种疫苗的小鼠将被提供给 RP2 进行 B 和 T 细胞分析。选定的候选疫苗（具有 接下来将在小鼠和叙利亚仓鼠中测试它们提供保护的能力 针对不同的冠状病毒。对于提供广泛保护（与对照相比）的候选疫苗，我们 还将评估免疫反应的持久性，以及疫苗接种对病毒传播的影响。 此外，这些候选疫苗将在 mRNA 脂质纳米颗粒疫苗平台中进行测试，该平台由 第一三共制药公司。