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）：提出的研究的总体目的是使用一种新型的自组装纳米颗粒包装方法来创建一种新的RNA病毒疫苗，旨在克服安全性，成本和其他限制，从而缓慢疫苗的发展。基于病毒的疫苗具有刺激先天和适应性免疫的能力，与其他疫苗输送方法相比，可以提高疫苗效力。与DNA病毒疫苗相比，基于RNA病毒的疫苗的安全性提高了，因为它们无法整合到宿主基因组中，但仍受到昂贵且限制疫苗促进的基于细胞的衣壳包装方法的限制。我们的初步数据表明，通过删除所有天然的衣壳组件约束，可以通过提高安全性，疫苗稳定性和非常低的成本来制造自组装疫苗，只需将RNA和涂层蛋白混合在一起即可。 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.最终的产物是一种病毒RNA疫苗，其包装特性与其天然衣壳无关，同时保留了使其成为良好疫苗抗原的病毒特征。其中包括免疫细胞最佳抗原摄取的NM颗粒大小，室温多年的稳定性以及安全地将病毒转基因表达携带到免疫细胞中的能力而没有病毒重建的风险。羊群病毒被选为自组装，因为它也具有许多理想的特征。它不是哺乳动物的病原体，因此将表现出改善的安全性。它在哺乳动物细胞类型中具有很高的抗原表达，与当前在疫苗发育中使用的RNA病毒相比，细胞死亡大大降低。羊群病毒先前尚未用于疫苗发育，因为天然的衣壳包装特性显着限制了转基因插入物尺寸，并且在细胞培养中仍需要制造病毒颗粒。该应用的预期结果将是克服天然的衣壳包装限制，并基于羊群内部病毒基因组创建自组装的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建具有成本效益，安全和强大的RNA疫苗，这是创新的，因为我们将验证任何具有理想特征的RNA病毒都可以适用于纳米颗粒自组装，并提高RNA疫苗开发的节奏以供人类使用。