Adaptation of a Novel RNA virus for vaccine use

新型RNA病毒用于疫苗用途的改造

基本信息

批准号：
8415821
负责人：
Alison Anne McCormick
金额：
$ 7.24万
依托单位：
TOURO UNIVERSITY OF CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8415821
关键词：
Adoption Alphavirus Alphavirus Infections Antibodies Antigens Apoptosis Attenuated Attenuated Vaccines Capsid Capsid Proteins Cell Culture Techniques Cell Death Cell Line Cells Characteristics Clinical Collection Communicable Diseases Communities Complex DNA Viruses DNA biosynthesis Data Dendritic Cells Development Drug Formulations Exhibits Functional RNA Future Genes Genetic Transcription Genome Genomics Goals Hour Housing Human Human Virus Immune Immunologic Surveillance In Vitro Insecta Kinetics Malignant Neoplasms Mammalian Cell Medical Methods Modification Natural Immunity Outcome Particulate Phase Phosphate Buffer Play Production Proteins Public Health RNA RNA Viruses RNA-Directed RNA Polymerase Reporter Research Research Priority Risk Role Safety Scientist Semliki forest virus Small RNA Surface System Temperature Testing Transfection Transgenes Vaccine Antigen Vaccines Viral Viral Genome Virion Virus Work adaptive immunity attenuation base cell type cost cost effective design human disease immune activation immunogenicity improved innovation member nanoparticle novel novel vaccines particle pathogen prevent promoter reconstitution response self assembly transgene expression uptake vaccine delivery vaccine development vaccine safety vector vaccine viral RNA virus characteristic

项目摘要

DESCRIPTION (provided by applicant): The overall objective of the research proposed is to create a new RNA virus vaccine using a novel self-assembling nanoparticle packaging method, designed to overcome safety, cost and other limitations that slow vaccine development. Virus-based vaccines have the ability to stimulate both innate and adaptive immunity, which can improve vaccine potency compared to other methods of vaccine delivery. RNA virus based vaccines have improved safety compared to DNA virus vaccines because they can't integrate into the host genome, but are still limited by cell-based capsid packaging methods that are costly and limit vaccine boosting. Our preliminary data show that by removing all native capsid assembly constraints self-assembling vaccines can be made with improved safety, vaccine stability, and at very low cost, simply by mixing RNA and coat protein together. The Specific Aims of this research plan are 1) to modify the insect Flock House Virus RNA genome with a non-native capsid origin of assembly and confirm in vitro particle formation, 2) to insert a foreig reporter transgene under the control of a virus promoter and evaluate the protein accumulation in cells, and 3) to combine the transgene expression and nanoparticle self-assembly, and to confirm that both are functional. The resulting product is a viral RNA vaccine with packaging characteristics independent of its native capsid, while retaining the virus characteristics that make it a good vaccine antigen. That includes nM particulate size for optimal antigen uptake by immune cells, stability at room temperature for years, and the ability to safely carry viral transgene expression into immune cells without the risk of virus reconstitution. Flock House Virus was selected for self-assembly because it also has many desirable characteristics. It is not a mammalian pathogen, and thus will exhibit improved safety. It has very high levels of antigen expression in mammalian cell types, and greatly reduced cell death compared to RNA viruses currently used in vaccine development. Flock House Virus has not been previously exploited for vaccine development because the native capsid packaging characteristics significantly limit transgene insert size and the virus particles still need to be made in cell culture. The expected outcome of this application will be to overcome native capsid packaging limitations, and create a self-assembling RNA nanoparticle based on the Flock House Virus genome, with high level antigen expression. Our research is significant, because it will fulfill our long-term objective to create cost effective, safe and robust RNA vaccines, and it is innovative because we will validate that any RNA virus with desirable characteristics can be adapted for nanoparticle self- assembly, and increase the pace of RNA vaccine development for human use.

描述（由申请人提供）：所提议研究的总体目标是使用新型自组装纳米颗粒包装方法创建一种新的RNA病毒疫苗，旨在克服安全性、成本和其他阻碍疫苗开发的限制。基于病毒的疫苗能够刺激先天免疫和适应性免疫，与其他疫苗递送方法相比，可以提高疫苗的效力。与 DNA 病毒疫苗相比，基于 RNA 病毒的疫苗提高了安全性，因为它们无法整合到宿主基因组中，但仍然受到基于细胞的衣壳包装方法的限制，这种方法成本高昂，并且限制了疫苗的加强。我们的初步数据表明，通过消除所有天然衣壳组装限制，只需将 RNA 和外壳蛋白混合在一起，就可以以非常低的成本制造出具有更高安全性和疫苗稳定性的自组装疫苗。该研究计划的具体目标是 1) 用非天然衣壳组装起源修改昆虫羊群病毒 RNA 基因组并确认体外颗粒形成，2) 在病毒启动子的控制下插入外来报告基因转基因并评估细胞中蛋白质的积累，3) 将转基因表达和纳米粒子自组装结合起来，并确认两者均具有功能。由此产生的产品是一种病毒 RNA 疫苗，其包装特性独立于其天然衣壳，同时保留了使其成为良好疫苗抗原的病毒特性。这包括实现免疫细胞最佳抗原摄取的 nM 颗粒大小、在室温下多年的稳定性以及将病毒转基因表达安全地携带到免疫细胞中而不存在病毒重建风险的能力。选择羊群病毒进行自组装是因为它还具有许多理想的特性。它不是哺乳动物病原体，因此将表现出更高的安全性。它在哺乳动物细胞类型中具有非常高水平的抗原表达，并且与目前用于疫苗开发的RNA病毒相比，大大减少了细胞死亡。羊群病毒之前尚未用于疫苗开发，因为天然衣壳包装特性显着限制了转基因插入物的大小，并且病毒颗粒仍然需要在细胞培养物中制备。该应用的预期结果将是克服天然衣壳包装限制，并创建基于羊群病毒基因组的自组装 RNA 纳米颗粒，具有高水平的抗原表达。我们的研究很重要，因为它将实现我们的长期目标创造具有成本效益、安全和稳健的 RNA 疫苗，它是创新的，因为我们将验证任何具有所需特性的 RNA 病毒都可以适应纳米颗粒自组装，并加快人类使用的 RNA 疫苗的开发步伐。