Development of a novel replicating viral RNA vaccine platform

项目摘要 病毒复制子，或自我复制的RNA元件不能启动传播感染， 传统上包装成病毒颗粒，称为病毒复制子颗粒（VRP），用于体内 交付.或者，它们可以在质粒DNA上编码并转染宿主细胞， 核介导的RNA复制的启动。然而，抗载体免疫与 关于前者，或后者方法中的基因组整合限制了 这样的技术。此外，VRP的发展仅限于较大的病毒 能够携带额外的基因货物。随着交付的改进， 裸RNA，现在有机会开发新的病毒复制子，而不受限制 由传统的交付模式提供。通过利用来自不同的阳性- 链RNA病毒家族，我们可以利用宿主使用的各种机制来检测 复制病毒RNA以及免疫逃逸的差异机制所利用的 亲本病毒，合理设计疫苗。我们提出的研究将阐明最小的 来自一种小的正链RNA动物病毒的遗传元件， 从亚基因组转录物复制和表达异源基因。然后我们将 展示Zika疫苗开发的转化应用， 肺结核、病毒和细菌性疾病，对公共卫生造成严重后果。 此外，我们还将对该平台与现有平台进行比较研究 这些技术来自于其他正链RNA病毒家族， 将引起不同的先天性和适应性免疫反应，由于进化的差异， 亲本病毒

PROJECT SUMMARY Viral replicons, or self-replicating RNA elements incapable of initiating a spreading infection, are traditionally packaged into virus particles, termed viral replicon particles (VRPs), for in vivo delivery. Alternatively, they can be encoded on a plasmid DNA and transfected for host cell nucleus-mediated launch of replicating RNA. However, concerns of anti-vector immunity with respect to the former, or genome integration in the latter approach has limited the clinical use of such technologies. Furthermore, the development of VRPs has been restricted to larger viruses that are capable of harboring additional genetic cargo. With improvements in the delivery of naked RNA, there are now opportunities to develop new viral replicons without the constraints provided by traditional delivery paradigms. By utilizing replicons derived from diverse positive- strand RNA virus families, we can exploit the various mechanisms used by the host to detect replicating viral RNA as well as the differential mechanisms of immune evasion utilized by the parental virus, to rationally design vaccines. We propose studies that will elucidate the minimal genetic elements derived from a small positive-stranded RNA animal virus that are required to replicate and express a heterologous gene from a subgenomic transcript. We will then demonstrate the translational applications towards vaccine development for Zika and tuberculosis, viral and bacterial diseases with significant public health consequences. Furthermore, we will perform comparative studies between this platform with existing platform technologies derived from other positive-strand RNA virus families with the hypothesis that each will elicit distinct innate and adaptive immune responses due to evolutionary differences of the parental viruses.