Rational design and evaluation of novel mRNA vaccines against MERS-CoV

针对 MERS-CoV 的新型 mRNA 疫苗的合理设计和评估

• 批准号: 10410839
• 负责人: Lanying Du
• 金额: $ 55.5万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10410839
• 关键词: Address; Antibodies; Antibody Response; Antigen Targeting; B cell repertoire; B-Lymphocytes; Bypass; Cells; Chromosomes; Communicable Diseases; Complex; Consumption; Coronavirus Infections; Coronavirus spike protein; Cytoplasm; Development; Dipeptidyl Peptidases; Disease Outbreaks; Dose; Ebola; Ebola virus; Evaluation; Formulation; Future; Generations; Genetic Transcription; Goals; Guidelines; Human; Immune; Immune response; Immunity; Immunization; Immunize; In Vitro; Industrialization; Influenza; Injections; Light; Lipids; Measures; Membrane Fusion; Messenger RNA; Microfluidics; Middle East Respiratory Syndrome; Middle East Respiratory Syndrome Coronavirus; Modeling; Modification; Mus; Nuclear Envelope; Nucleosides; Pathogenicity; Pattern; Production; Proteins; Public Health; RNA Stability; RNA vaccine; Regimen; Risk; SARS coronavirus; Safety; Subunit Vaccines; Technology; Testing; Time; Transgenic Organisms; Translations; Vaccination; Vaccine Production; Vaccines; Viral; Virus; Virus Diseases; Wild Type Mouse; Zika Virus; base; coronavirus vaccine; cost; cost effective; design; immunogenicity; improved; in vivo; in vivo evaluation; lipid nanoparticle; meetings; neutralizing antibody; new technology; next generation; next generation sequencing; nonhuman primate; novel; novel vaccines; pandemic disease; pathogen; pathogenic virus; protective efficacy; receptor binding; response; success; vaccine access; vaccine candidate; vaccine development; vaccine evaluation

Abstract Traditional strategies of vaccine development suffer from long-term and costly manufacture, and as a result, often fail to respond rapidly to newly emerging and reemerging infectious diseases. By contrast, messenger RNA (mRNA) is rising as a new technology platform to develop vaccines “on demand” against viral pathogens, offering attractive advantages such as cell-free production, non-viral delivery, as well as simple, fast and cost- effective manufacture. Further improvement upon mRNA's stability and translation efficiency, understanding of their immune mechanisms, and evaluation of their protective efficacy will facilitate the development of next- generation mRNA vaccine technologies against diverse viral pathogens. Middle-East respiratory syndrome (MERS) coronavirus (MERS-CoV) is a highly pathogenic, emerging infectious virus posing a continuous threat to public health worldwide. There are currently no MERS vaccines approved for use in humans. MERS-CoV spike (S) protein, particularly its receptor-binding domain (RBD), is an important vaccine target. We have previously shown that MERS-CoV RBD contains a critical neutralizing domain capable of inducing strong cross-neutralizing antibodies and protecting human dipeptidyl peptidase 4-transgenic (hDPP4-Tg) mice against MERS-CoV infection with outstanding efficacy. However, production of subunit vaccines and other traditional vaccines has limitations, such as low expression and complex purification. To address these unmet challenges, we propose to rationally design and evaluate novel mRNA vaccines, using MERS-CoV as a model pathogen and MERS-CoV S protein as a target antigen. We hypothesize that with appropriate modification and optimization, MERS-CoV S protein RBD-based mRNA vaccines will demonstrate improved stability, increased translation efficiency, and enhanced immunogenicity in both mouse and non-human primates (NHP) models, with protective efficacy on par with the RBD-based subunit vaccine. The specific aims are to (1) rationally design MERS-CoV mRNA vaccines with improved stability and translation efficiency, (2) carefully optimize mRNA formulations and immunization regimens towards in-vivo evaluation of their immunogenicity and mode of action in wild-type mice, and (3) comprehensively evaluate protective efficacy of MERS-CoV mRNA vaccines and elucidate their protective mechanisms in hDPP4-Tg mice and NHPs. Of note, we will also examine the utility of new technologies such as microfluidics and next-generation sequencing (NGS) analysis of B-cell response in mRNA vaccine development and evaluation. The long-term goal is to develop a safe and effective mRNA vaccine that is able to (1) maintain sufficient quantity and quality suitable for industrial- scale production, and (2) meet the WHO Target Product Profiles for rapid onset of immunity in outbreak settings and long-term protection of people at high ongoing risk of MERS-CoV. Together, the proposed project will shed light on protective mechanisms of mRNA vaccines, and provide much-needed information and guidelines for developing mRNA vaccines against diverse viral pathogens with pandemic potential.

抽象的 传统的疫苗开发策略面临着生产周期长且成本高昂的问题，因此， 相比之下，信使往往无法对新出现和再次出现的传染病做出快速反应。 RNA (mRNA) 正在成为一种新技术平台，可“按需”开发针对病毒病原体的疫苗， 提供有吸引力的优势，如无细胞生产、非病毒传递，以及简单、快速和成本低 进一步提高mRNA的稳定性和翻译效率，了解 它们的免疫机制以及对其保护功效的评估将促进下一代的开发 针对多种病毒病原体的新一代 mRNA 疫苗技术。 (MERS) 冠状病毒 (MERS-CoV) 是一种高致病性、新兴传染性病毒，构成持续威胁 目前还没有批准用于人类的 MERS 疫苗。 刺突（S）蛋白，特别是其受体结合域（RBD），是我们的重要疫苗靶标。 先前表明，MERS-CoV RBD 包含一个关键的中和结构域，能够诱导强 交叉中和抗体并保护人二肽基肽酶 4 转基因 (hDPP4-Tg) 小鼠免受 然而，与其他传统亚单位疫苗的生产相比，MERS-CoV感染具有突出的疗效。 疫苗有其局限性，例如低表达和复杂的纯化，以解决这些未满足的问题。 挑战，我们建议以 MERS-CoV 为模型，合理设计和评估新型 mRNA 疫苗 我们勇敢地面对病原体和 MERS-CoV S 蛋白作为目标抗原，并进行了适当的修改。 和优化，基于 MERS-CoV S 蛋白 RBD 的 mRNA 疫苗将表现出更高的稳定性， 提高小鼠和非人灵长类动物 (NHP) 的翻译效率并增强免疫原性 模型，其保护功效与基于 RBD 的亚单位疫苗相当，具体目标是 (1) 合理设计MERS-CoV mRNA疫苗，提高稳定性和翻译效率，（2）仔细 优化 mRNA 配方和免疫方案，以体内评估其免疫原性 以及在野生型小鼠中的作用方式，以及（3）综合评价MERS-CoV的保护功效 mRNA 疫苗并阐明其在 hDPP4-Tg 小鼠和 NHP 中的保护机制。 检查微流体和下一代测序 (NGS) 分析等新技术的实用性 mRNA 疫苗开发和评估中 B 细胞反应的长期目标是开发安全的疫苗。 有效的 mRNA 疫苗能够 (1) 保持足够的数量和质量，适合工业化- 规模生产，(2) 满足世界卫生组织目标产品概况，在疫情爆发时快速产生免疫力 拟议项目将环境和对中东呼吸综合征冠状病毒持续高风险人群的长期保护结合在一起。 将阐明 mRNA 疫苗的保护机制，并提供急需的信息和 针对具有大流行潜力的多种病毒病原体开发 mRNA 疫苗的指南。