Development of Viral Vaccines against Sarbecoviruses and Merbecoviruses

项目摘要 在过去的二十年中，六种冠状病毒（CoV）已经出现或扩大了它们的地理范围。的 对全球影响最大的新出现的人类冠状病毒（SARS-CoV-1、SARS-CoV-2和MERS-CoV）属于 冠状病毒科的β冠状病毒属内的Sarbecovirus和Merbecovirus亚属。 许多人畜共患的高风险Sarbecoviruses和Merbecoviruses准备为人类出现事件，因为 它们可以结合人ACE 2和DPP 4进入受体并感染培养中的人细胞。从历史 冠状病毒的爆发，再加上最近出现的SARS-CoV-2及其对人类的不稳定影响， 为了保护全球健康和经济，迫切需要开发能够广泛保护 现有和未来的Sarbecoviruses和Merbecoviruses。因此，P01提案的首要目标是 产生病毒载体疫苗，其诱导广泛的交叉保护性体液和细胞免疫， 具有大流行潜力的Sarbecoviruses和Merbecoviruses，特别是具有人畜共患病高风险的病毒 出现在人群中。项目3将使用项目1和项目2的抗原和表位设计， 创建疫苗平台，产生针对Sarbecoviruses的交叉反应性免疫应答， 具有大流行潜力的默贝科病毒。我们将使用几种刺突（S）、RBD和非结构蛋白抗原 其可以作为多价制剂的一部分施用以诱导广泛的B和T细胞免疫。项目3 将对鼻内递送的黑猩猩腺病毒载体病毒进行平行和迭代工程 （ChAd）和肌内递送的减毒活水疱性口炎病毒（VSV），所述减毒活水疱性口炎病毒（VSV）展示或产生 优化抗原抗原和疫苗，显示免疫反应的最高幅度和最大的 交叉反应性（用核心A测定）将在未处理、病毒免疫和mRNA接种小鼠中进行检测 为了确定对SARS-CoV-2的预先存在的免疫力如何影响我们更广泛的免疫原性， 针对CoV疫苗。显示最佳B和T细胞免疫原性的疫苗（宽度、幅度和免疫原性） 功能）将优先用于核心B中使用多种Sarbecovirus的小鼠和仓鼠攻毒研究， 梅尔贝科病毒我们的提案是产品开发的原理验证。我们设想至少产生 一种疫苗，诱导广谱免疫多种冠状病毒的关注。

Project Summary Over the past twenty years, six coronaviruses (CoV) have emerged or expanded their geographic range. The emergent human CoVs with the greatest global impact (SARS-CoV-1, SARS-CoV-2, and MERS-CoV) belong to the Sarbecovirus and Merbecovirus subgenera within the Betacoronavirus genus of the Coronaviridae family. Many zoonotic high-risk Sarbecoviruses and Merbecoviruses are poised for human emergence events because they can bind human ACE2 and DPP4 entry receptors and infect human cells in culture. Given the historical outbreaks of CoVs, coupled with the recent emergence of SARS-CoV-2 and its destabilizing consequence on global health and economy, there is an urgent need to develop vaccines capable of broad protection against existing and future Sarbecoviruses and Merbecoviruses. Thus, the overarching goal of this P01 proposal is to generate viral-vectored vaccines that induce broad cross-protective humoral and cellular immunity to Sarbecoviruses and Merbecoviruses with pandemic potential, especially those viruses at high risk for zoonotic emergence into human populations. Project 3 will use antigen and epitope designs from Projects 1 and 2 to create vaccine platforms that generate cross-reactive immune responses against Sarbecoviruses and Merbecoviruses with pandemic potential. We will use several spike (S), RBD, and non-structural protein antigens that can be administered as part of a polyvalent formulation to induce broad B and T cell immunity. Project 3 will perform parallel and iterative engineering of an intranasally delivered chimpanzee adenoviral vectored virus (ChAd) and an intramuscularly delivered live-attenuated vesicular-stomatitis virus (VSV) displaying or producing optimized antigens. Antigens and vaccines that show immune responses of the highest magnitude and greatest cross-reactivity (determined with Core A) will be tested in in naïve, virus-immune, and mRNA vaccinated mice to determine how pre-existing immunity to SARS-CoV-2 impacts the immunogenicity of our more broadly targeting CoV vaccines. Vaccines showing optimal B and T cell immunogenicity (breadth, magnitude, and function) will be prioritized for mouse and hamster challenge studies in Core B with multiple Sarbecoviruses and Merbecoviruses. Our proposal is a proof-of-principle for product development. We envision generating at least one vaccine that induces broad-spectrum immunity to multiple CoV of concern.