Epitope-Based Design and Modified RNA Platform for Bivalent Marburgvirus Vaccine

项目总结/摘要 马尔堡病毒（MARV）和拉文病毒（RAVV），它们都属于马尔堡病毒科的属 丝状病毒科引起人类的严重疾病，病死率高达90%。没有许可证 马尔堡病毒疫苗。针对埃博拉病毒（EBOV）的候选疫苗临床试验， 属于丝状病毒科的埃博拉病毒属，证明了高疫苗剂量， 在保护水平诱导免疫应答所需的药物导致与其 主要成分EBOV糖蛋白（GP）。我们和其他人最近分离并鉴定了人类 MARV和EBOV的单克隆抗体（mAb），并确定了MARV和EBOV的主要抗原决定簇。 中和和保护丝状病毒GP。我们已经成功地保护了非人类 通过被动转移的单克隆抗体对MARV的灵长类动物。这项研究的中心假设是， 来自MARV感染的幸存者的天然存在的人保护性抗体可以用作 为最佳的合理设计的疫苗结构的模板。我们最近的研究结果支持了这一假设。 从幸存者中分离保护性mAb的广泛进展，以及 计算免疫学技术。提出了基于结构的疫苗MARV的合理设计 在GP上呈现免疫原性决定簇的候选物。与野生型（wt）GP一样，抗原元件 蛋白质上的蛋白质被糖基化、聚糖帽和粘蛋白样结构域掩盖。所设计的 预期抗原比野生型GP更好地呈现保护性决定簇。我们还建议， 基于保守的GP表位的疫苗将对MARV和RAVV都具有保护性。该疫苗将 使用高度创新的疫苗递送平台，基于脂质中递送的假尿苷修饰的RNA 纳米颗粒制剂。该提案基于由不同专家组成的跨学科方法 在计算建模，抗体和抗原发现，丝状病毒病毒学，免疫学和疫苗学。 Meiler计算小组将使用ROSETTA软件平台技术设计新颖的结构， 基于疫苗的候选抗原，使用GP中抗原-抗体复合物的高分辨率结构 受体结合域。Crowe实验室将生产重组抗原和抗体， 构建体的适当结构和功能，并确定其生物分子相互作用的细节。 Moderna Therapeutics将提供创新的mRNA疫苗平台。布克列耶夫和盖斯伯特 实验室将在啮齿动物和非人类中测试表达设计抗原的疫苗构建体 灵长类动物模型的马尔堡病毒，和Bukreyev实验室将进行深入的表征， 免疫反应这项提案的完成将导致制定一个普遍和安全的下一个- 第二代疫苗，它将对MARV和RAVV都有保护作用。产生的抗原将是 与目前在临床试验中的任何现有先进疫苗平台兼容。

PROJECT SUMMARY/ABSTRACT The Marburg virus (MARV) and Ravn virus (RAVV), which both belong to the genus Marburgvirus of the family Filoviridae, cause the severe disease in humans, with case fatality rates up to 90%. There are no licensed vaccines against marburgviruses. Clinical trials of vaccine candidates against Ebola virus (EBOV), which belongs to the genus Ebolavirus of the family Filoviridae, demonstrated that the high vaccine doses that are required to induce an immune response at the protective level result in toxic effects associated with their principal component EBOV glycoprotein (GP). We and others have recently isolated and characterized human monoclonal antibodies (mAbs) to MARV and EBOV and defined the principal antigenic determinants for neutralization and protection on filovirus GP. We have demonstrated the successful protection of non-human primates against MARV by passively transferred mAbs. The central hypothesis of this study is that the epitopes of naturally-occurring human protective antibodies from survivors of a MARV infection can be used as templates for optimal rationally-designed structure-based vaccines. This hypothesis is supported by our recent extensive progress in the isolation of protective mAbs from survivors in conjunction with the recent advances in computational immunology techniques. We propose the rational design of structure-based vaccine MARV candidates that present the immunogenic determinants on GP. As in the wild-type (wt) GP, antigenic elements on the protein are obscured by glycosylation, the glycan cap and the mucin-like domain. The designed antigens are expected to better present protective determinants than the wt GP. We also propose that a vaccine based on conserved GP epitopes will be protective against both MARV and RAVV. The vaccine will use a highly innovative vaccine delivery platform based on pseudouridin-modified RNA delivered in a lipid nanoparticle formulation. The proposal is based on an interdisciplinary approach with a diverse team of experts in computational modeling, antibody and antigen discovery, filovirus virology, immunology and vaccinology. The Meiler computational group will use the ROSETTA software platform techniques to design novel structure- based vaccine candidate antigens, using high-resolution structures of antigen-antibody complexes in the GP receptor-binding domain. The Crowe laboratory will generate recombinant antigens and antibodies, validate proper structure and function of the constructs, and determine the fine details of their biomolecular interaction. Moderna Therapeutics will provide the innovative mRNA vaccine platform. The Bukreyev and Geisbert laboratories will test the vaccine constructs expressing the designed antigens in rodent and non-human primate models of marburgviruses, and the Bukreyev laboratory will perform in-depth characterization of the immune response. The completion of this proposal will result in the development of a universal and safe next- generation vaccine, which will be protective against both MARV and RAVV. The generated antigen will be compatible with any existing advanced vaccine platform currently in clinical trials.