Development of Universal Influenza Virus Vaccines Using Nucleoside-Modified Messenger RNA

使用核苷修饰信使 RNA 开发通用流感病毒疫苗

• 批准号: 10404632
• 负责人: Norbert Pardi
• 金额: $ 61.37万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-21 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10404632
• 关键词: Adverse event; Affinity; Animals; Antibodies; Antibody Affinity; Antibody Response; Antigens; B-Lymphocytes; Birds; Cell Culture Techniques; Cellular Assay; Cellular Immunity; Cessation of life; Clinical Trials; Combined Vaccines; Communicable Diseases; Data; Dendritic Cells; Development; Disease Outbreaks; Distant; Elements; Encapsulated; Ferrets; Ferritin; Generations; Gold; Grant; Head; Helper-Inducer T-Lymphocyte; Hemagglutinin; Human; Humoral Immunities; Immune; Immune response; Immunity; Immunization; Immunize; Immunoglobulin Class Switching; Immunoglobulin G; In Vitro; Influenza; Influenza A virus; Influenza B Virus; Injections; Investigation; Ion Channel; Knockout Mice; Length; Longevity; Macaca mulatta; Malignant Neoplasms; Membrane; Messenger RNA; Morbidity - disease rate; Mouse Strains; Mus; Neuraminidase; Nucleoproteins; Nucleosides; Pathogenicity; Physiological; Population; Production; Publishing; RNA vaccination; RNA vaccine; Reaction; Research; Serology test; Site; Skin; Standard Model; Structure of germinal center of lymph node; Transfection; Vaccination; Vaccine Production; Vaccine Research; Vaccines; Viral; Viral Antigens; Virus; Zika Virus; base; cost effective; design; draining lymph node; egg; fast protein liquid chromatography; genetic signature; immunogenicity; influenza infection; influenza virus vaccine; influenzavirus; lipid nanoparticle; long term memory; mortality; nanoparticle; neutralizing antibody; novel; novel vaccines; pandemic disease; programs; protective efficacy; protein purification; universal influenza vaccine; vaccination strategy; vaccine platform; vaccine trial

ABSTRACT Influenza virus is one of the most important human infectious diseases. The influenza mortality is estimated to be approximately 650,000 per year worldwide, in addition, occasional global pandemics can infect up to 20- 40% of the world's population. Licensed influenza virus vaccines require annual reformulation and readministration due to poor IgG longevity and lack of neutralization of related viruses. We have developed a new vaccine approach against influenza virus that uses antigen-encoding nucleoside-modified mRNA encapsulated in lipid nanoparticles (LNPs). This vaccine format composed entirely of physiologic components that is simple and cost effective to produce and should induce no adverse events after administration. Additionally, mRNA vaccine production is sequence-independent and does not require eggs, cell culture or protein purification. We demonstrated that monovalent mRNA-LNP vaccines encoding full-length hemagglutinin (HA) induced potent immune responses against the immunosubdominant HA stalk region, one of the targets of universal influenza virus vaccines, which was associated with protection against homologous, heterologous and heterosubtypic influenza viruses in mice. Furthermore, we showed that a single immunization with nucleoside-modified mRNA-LNP vaccines induced durable protective HA stalk-specific antibody responses. Finally, we demonstrated that nucleoside-modified mRNA-LNP immunization induced high levels of antigen-specific T follicular helper (Tfh) cells that are critical for the generation of germinal centers, immunoglobulin class switch, and long-term memory, which are vital elements in the development of an effective influenza vaccine. In this grant, we aim to develop and evaluate new generation, broadly protective influenza virus vaccines in mice and ferrets using nucleoside-modified mRNA-LNPs encoding hemagglutinin, neuraminidase, nucleoprotein and the ectodomain of M2. Additionally, we aim to investigate the mechanisms of action of mRNA-LNP influenza virus vaccines. We will have 3 specific aims: 1) Design and production of mRNA immunogens for conserved regions of influenza virus. 2) Immunogenicity and protective efficacy of nucleoside-modified mRNA-LNP influenza virus vaccines in mice and ferrets. 3) Investigation of the mechanisms of action of nucleoside-modified mRNA-LNP influenza vaccines. This proposal will develop safe, new generation influenza virus vaccines that are easy to manufacture and administer and can protect against antigenically distant influenza viruses. The data generated will be capable of moving this vaccine approach to clinical trial development.

抽象的 流感病毒是最重要的人类传染病之一。估计流感死亡率 此外，全球大约每年约650,000 全球40％的人口。许可的流感病毒疫苗需要年度重新审核，并且 由于IgG寿命差和相关病毒缺乏中和化而导致的读数。我们已经开发了 针对使用抗原编码核苷修饰mRNA的流感病毒的新疫苗方法 封装在脂质纳米颗粒（LNP）中。这种完全由生理成分组成的疫苗格式 这是简单且具有成本效益的生产，并且在给药后不应诱发不良事件。 此外，mRNA疫苗的产生是独立于序列的，不需要卵，细胞培养或 蛋白质纯化。我们证明了编码全长的单价mRNA-LNP疫苗 血凝素（HA）诱导针对免疫抑制剂HA茎区的有效免疫反应，一个 普遍流感病毒疫苗的靶标，这与防御同源性有关 小鼠的异源和异源性流感病毒。此外，我们证明了一个 用核苷改性的mRNA-LNP疫苗免疫诱导的耐用防护性HA茎特异性 抗体反应。最后，我们证明了核苷改性的mRNA-LNP免疫诱导的高 抗原特异性T卵泡辅助（TFH）细胞的水平，这对于生发中心的产生至关重要， 免疫球蛋白类开关和长期记忆是发展的重要因素 有效的流感疫苗。在这笔赠款中，我们旨在开发和评估新一代，广泛保护 使用核苷改性的mRNA-LNP，编码黑凝集素的核苷MRNA-LNP，流感病毒疫苗和雪貂 神经氨酸酶，核蛋白和M2的胞外域。此外，我们旨在研究机制 mRNA-LNP流感病毒疫苗的作用。我们将有3个具体目标：1）设计和生产 流感病毒保守区域的mRNA免疫原。 2）免疫原性和保护性 小鼠和雪貂中的核苷修饰的mRNA-LNP流感病毒疫苗。 3）调查 核苷改性mRNA-LNP流感疫苗的作用机理。该建议将发展安全， 易于制造和管理的新一代流感病毒疫苗 抗原远处的流感病毒。生成的数据将能够将这种疫苗方法移动到 临床试验开发。