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.

项目摘要 该提议是开发基于RNA的快速响应影响疫苗原型，支持Niaid的呼吁 用于预防大流行影响的核酸基疫苗。鸟类影响H7N9原因 严重的呼吸疾病，死亡率高。该病毒的高人畜共患能力引起了严重的关注 出于大流行的可能性，风险可能与H5菌株相似。虽然进度有 他是在开发H5影响力疫苗的过程中造成的，H7产品落后了。提出的H7N9疫苗 将在两个方面具有创新性。 1）H7N9疫苗通常专注于血凝素（HA）蛋白作为 主要免疫原，但要对HA H7进行免疫调查，虽然保护，但比其他人弱 在人类中。因此，TIBA将开发一种多抗原方法，特别是包括优化比率 Virion蛋白HA，NA，M1和NP。预计这种方法将增加免疫原性和异型 疫苗的保护潜力。 2）常规脂质纳米颗粒（LNP），这是核的主要 酸递送需要大量的“结构”脂质，从而导致相对较低的RNA含量。 Tiba有 开发了一种改良的树枝状聚合物纳米颗粒系统，可最大化递送的RNA质量含量，保护 从降解中的RNA，并可以在体内有效摄取。这里开发的原型组成 将作为一个可以迅速实施任何爆发抗原序列的平台。 Tiba会见面 开发原型HA/NA/M1/NP疫苗的近期目标，以迈向实时病毒挑战 通过完成三个阶段1的目标，在第2阶段进行了实验和辅助研究。首先是验证 在体内基于H7 HA复制RNA的表达构建体的性能。这将被生成和制定 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。这些研究将确定随后实验所需的最低剂量。 在第二个目标中，TIBA将生成和测试Na，M1和NP mRNA和副本的单个公式 候选人，通过T细胞ELISPOTS以类似的剂量验证其在体内的性能，也是NA的 ELISA测量抗体反应。最终目的是组成多抗原原型疫苗 通过共同包裹四个 有效载荷以单个纳米颗粒公式。这种多抗原候选疫苗将在男性和 雌性BALB/C小鼠，与仅HA的抗原编码对照疫苗相比 免疫原性是针对所有组件保留的，如果抗HA反应受益于包含 附加的病毒素蛋白。候选原型将与最先进的LNP公式进行比较 与当前的RNA疫苗的金标准相比，评估TIBA输送技术的性能。