(Not Active) Novel approach for development of live & inactivated viral vaccines

（不活跃）实时开发的新方法

• 批准号: 8260254
• 负责人: Aaron Cole Brault
• 金额: --
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8260254
• 关键词: Acute; Amino Acid Substitution; Attenuated; Cells; Characteristics; Codon Nucleotides; Country; Development; Directed Molecular Evolution; Disease; Emerging Communicable Diseases; Engineering; Genetic Recombination; Immune response; Infection; Infectious Diseases Research; Life; Mediating; Methodology; MicroRNAs; Modification; Pathology; Phenotype; Population; Production; Proteins; Regulation; Safety; Serial Passage; Time; Tissues; Translations; Uncertainty; Vaccines; Viral; Viral Genome; Viral Vaccines; Virus; Virus Diseases; attenuation; base; biodefense; design; improved; novel; novel strategies; pathogen; prophylactic; vaccine candidate; vaccine development; vaccine safety

Currently, no vaccines exist for a significant number of historically important viral pathogens and newly emerging viral zoonotic disease threats. In part, this situation results from the incompatibility of existing methodologies with the challenges posed by a wide array of emerging viruses. The extent of attenuation mediated by approaches such as serial passaging of viruses, viral recombination and directed molecular evolution is unpredictable and this characteristic contributes to the lengthy production times for many vaccines made using these strategies. Likewise, a number of vaccines constructed using these methodologies have demonstrated poor safety profiles due to the inherent uncertainty in the number of attenuation determinants introduced. In this study we aim to further define a novel strategy to intuitively construct live-attenuated viral (LAV) vaccines applicable to the development of vaccines to a broad array of acute viral diseases. Our approach is based on the manipulation of host strategies for the regulation of protein translation. We hypothesize that use of rare codons as attenuation determinants for vaccine candidates will produce more stably attenuated viruses that can be rationally engineered with predictable replication phenotypes within host cells. Moreover, we have demonstrated that these substitutions can be introduced into various places in viral genomes, unlike amino acid substitutions which are conventionally employed to attenuate viruses. We have also designed a second strategy to further improve the safety of vaccines that restricts the replication of vaccine viruses in tissues that are responsible for engendering disease pathology. By introducing the target sequences for tissue-specific miRNAs into the viral genome we have demonstrated the ability to specifically block infection of target cells expressing the cognate miRNA, while permitting normal unaltered levels of replication in the cell populations required for the induction of a protective immune response. Importantly, this strategy is compatible with the modification of existing vaccines to improve their safety profiles. The approaches defined in this study have broad implications for the rapid development of prophylactic agents to a variety of disease threats.

目前，对于大量重要的历史病毒病原体和新的疫苗 新兴的病毒性人畜共患病威胁。在某种程度上，这种情况是由于现有的不兼容而引起的 各种新兴病毒带来的挑战的方法论。衰减的程度 通过诸如病毒，病毒重组和定向分子等方法介导的 进化是不可预测的，这种特征有助于许多人的长期生产时间 使用这些策略进行的疫苗。同样，使用这些疫苗建造了许多疫苗 方法学表明，由于数量固有的不确定性，安全概况不佳 引入了衰减决定因素。在这项研究中，我们旨在进一步定义一种新颖的策略来直观 构建适用于开发疫苗到广泛阵列的实时病毒（LAV）疫苗 急性病毒疾病。我们的方法是基于操纵主机策略的监管 蛋白质翻译。我们假设使用稀有密码子作为疫苗的衰减决定因素 候选人将产生更稳定的减弱病毒，可以通过可预测的 宿主细胞中的复制表型。而且，我们已经证明了这些替代可能是 与氨基酸取代相比，被引入病毒基因组的各个地方 被用来减弱病毒。我们还设计了第二种策略，以进一步提高 限制了负责产生的组织中疫苗病毒复制的疫苗 疾病病理学。通过将组织特异性miRNA的靶序列引入病毒基因组中，我们已经证明了能够特异性地阻断表达同源miRNA的靶细胞的感染，同时允许在诱导保护性免疫反应所需的细胞群体中正常的未改变水平的复制水平。重要的是，该策略与现有疫苗改善其安全性概况的修改兼容。这项研究中定义的方法对预防剂的快速发展对各种疾病威胁具有广泛的影响。