Rapid response to pandemic influenza via multi-antigen RNA-based vaccine

通过基于多抗原 RNA 的疫苗快速应对大流行性流感

• 批准号: 10491533
• 负责人: Christian W. Mandl
• 金额: $ 28.49万
• 依托单位: TIBA BIOTECH, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-10 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10491533
• 关键词: Adjuvant; Amines; Antibody Response; Antigens; Avian Influenza; B-Lymphocytes; Behavior; Benchmarking; Biological Assay; CD69 antigen; Cell Culture Techniques; Cell surface; Cells; Code; Codon Nucleotides; Combined Vaccines; Control Groups; Dendrimers; Development; Disease Outbreaks; Dose; Dyes; Economics; Encapsulated; Ensure; Enzyme-Linked Immunosorbent Assay; Equilibrium; Evaluation; Exclusion; Female; Formulation; Gel; Generations; Goals; Gold; H7 hemagglutinin; Hemagglutinin; Human; Immune response; Immunologics; In Vitro; Inbred BALB C Mice; Individual; Influenza; Influenza A Virus, H7N9 Subtype; Influenza A virus; Lead; Lipids; Measures; Memory; Messenger RNA; Methods; Molecular; Mus; NP protein; National Institute of Allergy and Infectious Disease; Neutralization Tests; Nucleic Acid Vaccines; Nucleic Acids; Outcome; Performance; Phase; Preparation; Prevention; Proteins; RNA; RNA Degradation; RNA vaccine; Replicon; Ribonucleases; Risk; ST14 gene; Sepharose; Serum; Splenocyte; Stains; Structural Protein; System; T-Lymphocyte; Technology; Testing; Time; TimeLine; Vaccines; Validation; Virion; Virus; Zoonoses; antibody detection; arm; base; density; design; experimental study; expression vector; flexibility; high risk; immunogenic; immunogenicity; in vivo; influenza virus vaccine; innovation; interest; light scattering; lipid nanoparticle; male; mortality; mouse model; nanoparticle; novel; nucleic acid delivery; pandemic disease; pandemic influenza; potency testing; prototype; respiratory; response; scale up; seasonal influenza; uptake; vaccine candidate; vaccine development; vaccine formulation; vector

Project Summary The proposal is to develop an RNA-based, rapid-response influenza vaccine prototype, supporting NIAID’s call for nucleic acid-based vaccines that protect against pandemic influenza threats. Avian influenza A H7N9 causes severe respiratory illness with a high mortality rate. The virus’ high zoonotic capacity has raised serious concerns over the possibility of a pandemic, with the risk being potentially similar to that of H5 strains. While progress has been made in the development of H5 influenza vaccines, H7 products have lagged. The proposed H7N9 vaccine will be innovative in two respects. 1) H7N9 vaccines typically focus on the hemagglutinin (HA) protein as the main immunogen, but immune responses to HA H7, while protective, are weaker than those against other HAs in humans. Therefore, Tiba will develop a multi-antigen approach, specifically including optimized ratios of the virion proteins HA, NA, M1, and NP. This approach is expected to increase the immunogenicity and heterotypic protective potential of the vaccine. 2) Conventional lipid nanoparticles (LNPs), which are the mainstay of nucleic acid delivery, require a large proportion of “structural” lipid, resulting in a relatively low RNA content. Tiba has developed a modified dendrimer nanoparticle system that maximizes the delivered RNA mass content, protecting RNA from degradation, and enables efficient uptake by cells in vivo. The prototype composition developed here will serve as a platform into which any outbreak antigen sequences could be rapidly implemented. Tiba will meet the near-term goal of developing a prototype HA/NA/M1/NP vaccine to advance toward live virus challenge experiments and IND-enabling studies in Phase 2 by completing three Phase 1 Aims. The first is to validate performance of a H7 HA replicon RNA-based expression construct in vivo. This will be generated and formulated with Tiba’s delivery system and tested in BALB/c mice at 0.2 µg, 2.0 µg, and 20 µg to measure cellular and humoral immunogenicity. These studies will establish the minimum dose required for subsequent experiments. In the second Aim, Tiba will generate and test individual formulations of NA, M1, and NP mRNA and replicon candidates, validating their performance in vivo at similarly increasing doses by T cell ELISpots and, for NA, also by ELISA to measure antibody responses. The final Aim is to compose a multi-antigen prototype vaccine combining the optimal balance of HA replicon RNA with NA, M1, and NP RNAs by co-encapsulating the four payloads in a single nanoparticle formulation. This multi-antigen candidate vaccine will be tested in male and female BALB/c mice, in comparison to HA-only and irrelevant antigen-coding control vaccines to determine if immunogenicity is retained against all components, and if anti-HA responses benefit from inclusion of the additional virion proteins. The candidate prototypes will be compared to state-of-the-art LNP formulations to evaluate the performance of Tiba’s delivery technology compared to the current gold-standard for RNA vaccines.

项目概要 该提案旨在开发一种基于 RNA 的快速反应流感疫苗原型，支持 NIAID 的号召 用于预防大流行性流感威胁的核酸疫苗。甲型H7N9禽流感的病因 严重的呼吸道疾病，死亡率很高。该病毒的高人畜共患能力引起了严重关注 流感大流行的可能性，其风险可能与 H5 毒株相似。虽然取得了进展 在H5流感疫苗的研发上，H7产品已经滞后。拟议的 H7N9 疫苗 将会在两个方面进行创新。 1) H7N9 疫苗通常以血凝素 (HA) 蛋白为重点 主要免疫原，但对 HA H7 的免疫反应虽然具有保护作用，但比针对其他 HA 的免疫反应弱 在人类中。因此，Tiba 将开发一种多抗原方法，具体包括优化的比例 病毒体蛋白 HA、NA、M1 和 NP。这种方法有望增加免疫原性和异型性 疫苗的保护潜力。 2）传统的脂质纳米粒（LNPs），它是核酸的支柱 酸输送需要大量的“结构”脂质，导致 RNA 含量相对较低。蒂巴有 开发了一种改良的树枝状聚合物纳米颗粒系统，可最大限度地提高所传递的 RNA 质量含量，保护 RNA 免遭降解，并能够在体内被细胞有效摄取。此处开发的原型组合物 将作为一个平台，任何爆发的抗原序列都可以快速实施。蒂巴将会见面 开发原型 HA/NA/M1/NP 疫苗以推进活病毒挑战的近期目标 通过完成三个第一阶段目标，在第二阶段进行实验和 IND 支持研究。首先是要验证 基于 H7 HA 复制子 RNA 的表达构建体的体内性能。这将被生成并制定 使用 Tiba 的递送系统，并在 BALB/c 小鼠中以 0.2 µg、2.0 µg 和 20 µg 进行测试，以测量细胞和 体液免疫原性。这些研究将确定后续实验所需的最小剂量。 在第二个目标中，Tiba 将生成并测试 NA、M1 和 NP mRNA 和复制子的单独配方 候选者，通过 T 细胞 ELISpots 在类似增加的剂量下验证其体内性能，对于 NA，还 通过 ELISA 测量抗体反应。最终目标是构建多抗原原型疫苗 通过共封装四种 RNA，将 HA 复制子 RNA 与 NA、M1 和 NP RNA 的最佳平衡结合起来 单个纳米颗粒制剂中的有效负载。这种多抗原候选疫苗将在男性和女性中进行测试 雌性 BALB/c 小鼠，与仅使用 HA 的疫苗和不相关抗原编码的对照疫苗进行比较，以确定是否 针对所有成分均保留免疫原性，并且如果抗 HA 反应受益于包含 额外的病毒体蛋白。候选原型将与最先进的 LNP 配方进行比较，以 与当前 RNA 疫苗的黄金标准相比，评估 Tiba 递送技术的性能。