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疫苗是疫苗科学感兴趣的增长领域。他们很灵活，因为净化 从一种疫苗到另一种疫苗的过程应该相似，并且可以迅速做出响应 到新兴疾病。将这些核酸构建体用作疫苗平台有许多 优点：纯化相对精简，可以使用DNA在几天内建立RNA构建体 合成技术随后进行转录。这允许对新兴病原体的快速反应 制造业的威胁或枢轴变化是为了采用新的循环菌株。这些疫苗显示 伟大的希望，在某些情况下，它们在人体试验中缺乏效率，并且 - 像蛋白质疫苗一样 - 可能 需要一种增强其诱导自适应免疫回避能力的方法。我们在这里提出建议 初步研究可以使RNA的新调整分子启用高通量屏幕（HTS） 疫苗。 在拟议的研究结束时，我们打算将新化合物的体外筛选 可以（1）增强RNA构建体表达的类； （2）打开NF-κB以增强 这些构建体的免疫原性，同时保持表达水平。这项工作将在 两个目的：首先，我们将开发一个绿色荧光蛋白（GFP）的记者RNA屏幕 措施翻译蛋白质水平（AIM 1）。然后我们将开发并限定基于NF-κB 来自AIM 1的蛋白质表达屏幕顶部的记者屏幕（AIM 2）。 未来的研究将包括在高通量设置中部署这些测定和大量筛选 铅的库，增强RNA构建体表达水平和RNA疫苗的免疫原性 从而成为RNA疫苗接种的调节器。