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

项目摘要/摘要 马尔堡病毒(Marburg Virus，MARV)和Ravn病毒(Ravn Virus，RAVV)同属马尔堡病毒科 丝状病毒科引起人类的严重疾病，病死率高达90%。没有获得许可的 针对马尔堡病毒的疫苗。埃博拉病毒(EBOV)候选疫苗的临床试验， 属于丝状病毒科的埃博拉病毒属，表明高剂量的疫苗是 在保护性水平上诱导免疫反应所需的物质会导致与其相关的毒性效应 主成分EBOV糖蛋白(GP)。我们和其他人最近分离并描述了人类 抗MARV和EBOV的单抗，并确定了主要的抗原决定因素 丝状病毒GP的中和与保护。我们已经展示了对非人类的成功保护 灵长类动物通过被动转移单抗对抗MARV。这项研究的中心假设是表位 来自MARV感染幸存者的自然产生的人类保护性抗体可以用作 最佳合理设计的基于结构的疫苗的模板。这一假设得到了我们最近 在从幸存者身上分离保护性单抗方面取得了广泛进展，同时也取得了最新进展 计算免疫学技术。我们提出了基于结构的疫苗Marv的合理设计 提出GP免疫原性决定簇的候选人。与野生型(Wt)gp一样，抗原元件 蛋白质上的糖基化、糖链和粘蛋白样结构域被遮盖。设计的 与wt gp相比，抗原有望更好地呈现保护性决定簇。我们还建议一项 基于保守gp表位的疫苗将对Marv和RAVV具有保护作用。疫苗将会 使用高度创新的疫苗递送平台，该平台基于以脂质形式递送的伪尿苷修饰的RNA 纳米颗粒配方。该提案基于一种跨学科的方法，由不同的专家团队组成 在计算建模、抗体和抗原发现、丝状病毒病毒学、免疫学和疫苗学中。 迈勒计算小组将使用Rosetta软件平台技术来设计新的结构- 基于疫苗候选抗原，利用GP中抗原-抗体复合体的高分辨结构 受体结合域。克劳实验室将产生重组抗原和抗体，验证 正确的结构和功能，并确定其生物分子相互作用的细节。 莫德纳治疗公司将提供创新的信使核糖核酸疫苗平台。布克雷耶夫和盖斯伯特 实验室将在啮齿动物和非人类身上测试表达设计抗原的疫苗构建物 马尔堡病毒的灵长类模型，Bukreyev实验室将对 免疫反应。这项建议的完成将导致制定一项普遍和安全的下一步- 一代疫苗，它将对Marv和RAVV具有保护作用。产生的抗原将是 与目前处于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.