Project 2: Structure-based antigen and nanoparticle vaccine design

项目摘要-项目2：基于结构的抗原和纳米颗粒疫苗设计 项目2的重点是新型稳定抗原的设计、生产和表征，以及广泛的 保护性纳米颗粒疫苗。在目标1中，我们将建立自动计算管道来识别 各种肉瘤病毒、多角体病毒和多角体病毒的刺突蛋白中的一组新的融合稳定突变 我们将在我们的广泛保护性疫苗中使用肺炎病毒株。我们最近的工作也证明了 利用深层突变扫描数据指导分离株稳定突变的鉴定 SARS-CoV-2受体结合域(RBD)。我们开发的计算管道将扩展这一点 在所有已知的四种肉瘤病毒分支中设计额外的稳定的RBD抗原的方法 用于泛肉瘤病毒疫苗。在目标2中，我们将通过计算设计新型的自组装蛋白质 以专门设计的阵列呈现预融合尖峰三聚体的纳米颗粒支架 不受限制的B细胞受体/抗体获得广泛保守的表位。我们假设这些 支架将引发具有超强保护广度的抗体反应，特别是当用作 马赛克纳米颗粒免疫原。我们将通过实验验证几种新的纳米粒子和原型 它们作为单价纳米颗粒免疫原在免疫和挑战研究中的功能表现 以确定最能将体液免疫反应集中在保守表位上的支架。在AIM 3中 我们将在我们最近的工作的基础上，展示广泛的保护性免疫反应的激发 通过产生显示多个冠状病毒rbd或预融合的多价纳米颗粒疫苗而引起的流感 斯派克斯。我们将采取层级的、系统发展驱动的方法。我们将首先专注于开发一种 泛肉瘤病毒疫苗，它将显示多个RBD或预融合尖峰抗原。然后我们将生成 具有广泛保护性的丝状病毒和鸡传染性支气管炎病毒疫苗显示出多个灌流高峰。最后，我们会 确定泛贝塔冠状病毒纳米颗粒疫苗的最佳成分。我们将与 我们计划中的其他小组确定主要的泛肉瘤病毒和泛贝塔冠状病毒候选疫苗 在我们计划的第二年和第五年结束时，进一步的临床前和临床发展。

PROJECT SUMMARY – PROJECT 2: Structure-based antigen and nanoparticle vaccine design Project 2 is focused on the design, production, and characterization of novel stabilized antigens and broadly protective nanoparticle vaccines. In Aim 1, we will establish automated computational pipelines that identify sets of novel prefusion-stabilizing mutations in the spike proteins of various sarbecovirus, merbecovirus, and embecovirus strains we will use in our broadly protective vaccines. Our recent work has also demonstrated the utility of using deep mutational scanning data to guide the identification of stabilizing mutations in isolated SARS-CoV-2 receptor-binding domains (RBD). The computational pipelines we develop will extend this method to the design of additional stabilized RBD antigens across all four known clades of sarbecoviruses for use in pan-sarbecovirus vaccines. In Aim 2, we will computationally design novel self-assembling protein nanoparticle scaffolds that present prefusion spike trimers in arrays specifically designed to allow unconstrained B cell receptor/antibody access to broadly conserved epitopes. We hypothesize that these scaffolds will elicit antibody responses with superior protective breadth, particularly when used as the basis for mosaic nanoparticle immunogens. We will experimentally validate several novel nanoparticles and prototype their functional performance as monovalent nanoparticle immunogens in immunization and challenge studies in mice to identify the scaffolds that best focus the humoral immune response on conserved epitopes. In Aim 3 we will build on our recent work demonstrating the elicitation of broadly protective immune responses against influenza by generating multivalent nanoparticle vaccines displaying multiple coronavirus RBDs or prefusion spikes. We will take a hierarchical, phylogeny-driven approach. We will focus at first on developing a pan-sarbecovirus vaccine, which will display multiple RBD or prefusion spike antigens. We will then generate broadly protective merbecovirus and embecovirus vaccines displaying multiple prefusion spikes. Finally, we will define the optimal composition of a pan-betacoronavirus nanoparticle vaccine. We will work closely with the other groups in our Program to identify lead pan-sarbecovirus and pan-betacoronavirus vaccine candidates for further preclinical and clinical development at the end of Years 2 and 5 of our Program, respectively.