Developing a Screen for New Adjuvants to Enhance RNA Vaccines

开发增强 RNA 疫苗的新佐剂筛选方案

• 批准号: 10189513
• 负责人: DARRICK Albert CARTER
• 金额: $ 8.68万
• 依托单位: PAI LIFE SCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-12 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10189513
• 关键词: Address; Adjuvant; Adopted; Affect; Agonist; Antigens; Antiviral Agents; Area; Biological Assay; Cells; DNA biosynthesis; Data; Disease; Formulation; Future; Genetic Transcription; Green Fluorescent Proteins; Herpes zoster disease; Human; In Vitro; Interferons; Libraries; Measures; Methods; Modernization; Nucleic Acids; Outcome; Population; Process; Proteins; RNA; RNA delivery; RNA vaccine; Reporter; Reporting; Research; Sampling; System; Technology; Testing; Translating; Translations; Vaccination; Vaccine Adjuvant; Vaccines; Work; adaptive immune response; aluminum sulfate; base; emerging pathogen; flexibility; global health; high throughput screening; immunogenicity; improved; interest; multiplex assay; programs; protein expression; response; screening; uptake; vaccinology

PROJECT SUMMARY RNA vaccines are a growing area of interest in vaccinology. They are flexible in that the purification process should be similar from one vaccine to the other and that they can be rapidly made in response to emerging diseases. Use of these nucleic acid constructs as a vaccine platform has numerous advantages: Purification is relatively streamlined, and RNA constructs can be built in days using DNA synthesis technologies followed by transcription. This allows for rapid responses to emerging pathogen threats or pivot changes in manufacturing to adopt to new circulating strains. While these vaccines show great promise, in some cases they lack full efficacy in human trials and - like protein vaccines - may require a method of enhancing their ability to induce adaptive immune responses. We propose here preliminary studies to enable a high-throughput screen (HTS) for new adjuvant molecules for RNA vaccines. At the end of the proposed research we intend to have qualified an in vitro screen for new compound classes that can (1) enhance expression of RNA constructs; and (2) turn on NF-κB in order to enhance immunogenicity of these constructs while maintaining expression levels. The work will be performed in two Aims: First, we will Develop a Green Fluorescent Protein (GFP)-based reporter RNA screen that measures translated protein levels (Aim 1). And then we will Develop and qualify an NF-κB-based reporter screen on top of the protein expression screen from Aim 1 (Aim 2). Future studies will include deploying these assays in a high throughput setting and screening large libraries for leads that enhance expression level of RNA constructs and immunogenicity of RNA vaccines thereby becoming adjuvants for RNA vaccination.

项目摘要 RNA疫苗是疫苗学中越来越感兴趣的领域。它们的灵活性在于， 一种疫苗与另一种疫苗的生产过程应该是相似的， 到新出现的疾病。使用这些核酸构建体作为疫苗平台具有许多优点。 优点：纯化相对简化，使用DNA可以在几天内构建RNA构建体。 合成技术，然后转录。这允许对新出现的病原体做出快速反应 威胁或制造业的关键变化，以适应新的流通菌株。虽然这些疫苗显示 虽然前景广阔，但在某些情况下，它们在人体试验中缺乏完全的功效，而且--就像蛋白质疫苗一样--可能 需要一种增强其诱导适应性免疫应答的能力的方法。我们在此提议 初步研究，使高通量筛选（HTS）的新佐剂分子的RNA 疫苗。 在拟议的研究结束时，我们打算对新化合物进行体外筛选 可以（1）增强RNA构建体的表达;和（2）开启NF-κB以增强 这些构建体的免疫原性，同时维持表达水平。工作将在 两个目标：首先，我们将开发一种基于绿色荧光蛋白（GFP）的报告RNA筛选， 测量翻译的蛋白质水平（目标1）。然后我们将开发并鉴定一种基于NF-κ B的 在来自Aim 1（Aim 2）的蛋白质表达筛选之上的报告基因筛选。 未来的研究将包括在高通量环境中部署这些检测，并筛选大的 用于增强RNA构建体的表达水平和RNA疫苗的免疫原性的先导物的文库 从而成为RNA疫苗接种的佐剂。