Rapid Generation of Vaccine Candidates Against Novel Coronavirus (SARS-CoV-2) Using the Bacteriophage T4 Nanoparticle Platform

使用噬菌体 T4 纳米颗粒平台快速生成针对新型冠状病毒 (SARS-CoV-2) 的候选疫苗

基本信息

批准号：
10265803
负责人：
Richard J. Kuhn
金额：
$ 3.02万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-14 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10265803
关键词：
2019-nCoV Address Adjuvant Americas Animal Model Animal Testing Anthrax disease Antibodies Antigen-Presenting Cells Antigens Bacteriophage T4 Bacteriophages Basic Science Biological Assay COVID-19 vaccine CRISPR-nanoparticles Capsid Capsid Proteins Cardiovascular system Cells Cessation of life Chimeric Proteins China Cities Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Coronavirus Country E protein Engineering Enzyme-Linked Immunosorbent Assay Escherichia coli Future Generations Genes Genome Goals Human Immune Sera Immune response Immune system Immunization Immunize Individual Infection Intramuscular Laboratories Lead Macaca Medial Middle East Respiratory Syndrome Coronavirus Molecular Mus Muscle Cells Natural Immunity Nucleocapsid Proteins Oryctolagus cuniculus Pattern Plague Population Protein Binding Domain Proteins Province Rattus Receptor Gene Recombinants SARS coronavirus SARS-CoV-2 infection SARS-CoV-2 inhibitor Series Site Structure Surface Technology Testing Time Toll-like receptors Translational Research United States Vaccines Virion Virus adaptive immunity biodefense biothreat coronavirus vaccine design env Gene Products flexibility global health global health emergency immunogenicity mouse model nanoparticle novel novel coronavirus pandemic disease particle pathogen pathogenic virus promoter receptor binding response scaffold vaccine candidate vaccine delivery virtual

项目摘要

PROJECT SUMMARY This proposal aims to rapidly generate vaccine candidates against the 2019 novel coronavirus SARS-CoV- 2. Since its emergence about three months ago, this virus has caused more than 120,000 infections and 4,300 deaths worldwide and is rapidly spreading to virtually every country including the United States. This global health emergency must be immediately addressed by rapidly developing medial countermeasures. Our bacteriophage (phage) T4 vaccine platform is uniquely suited to address this threat. Developed in PI's laboratory, the T4 vaccines have been proven to generate robust humoral as well as cellular immune responses and confer complete protection against anthrax and plague in multiple animal models including mice, rats, rabbits, and macaques. The T4 vaccines do not need an adjuvant as its surface structure mimics the Pathogen- Associated Molecular Patterns (PAMPs) of viral pathogens and stimulate strong innate and adaptive immunity. The 120 x 86 nm phage T4 capsid is packaged with 171 kb genome and decorated with two non-essential outer capsid proteins; 870 molecules of Soc (small outer capsid protein) and 155 copies of Hoc (highly antigenic outer capsid protein). In specific aim 1, a series of T4-corona phages will be constructed by incorporating SARS- CoV-2 virion components individually and in combinations, by CRISPR engineering. The gene encoding the entire spike ectodomain will be inserted into phage genome under the control of the strong CAG promoter. Upon immunization, host cells (myocytes and antigen presenting cells at the site of immunization) take up phage particles and secrete the ectodomain trimers continuously, stimulating the immune system for weeks to months. The gene for the receptor binding domain (RBD) of S protein will be inserted such that the RBD will be expressed in host cells, as well as in E. coli as a Soc fusion protein which will then be displayed on phage capsid up to 870 copies per capsid. The ectodomain of E protein will be fused to Hoc and displayed up to 155 copies per capsid. Finally, ~400 copies of N protein will be packaged inside the capsid as part of the scaffolding core. In specific aim 2, the above T4-corona recombinant phages will be evaluated for elicitation of SARS-CoV- 2 virion-specific immune responses in a mouse model. Mice will be immunized with purified phage particles intramuscularly and the immune responses will be quantified by ELISA, competitive receptor binding, ELISpot, and virus neutralization assays. We expect that the T4-corona vaccines will elicit robust antibody and cellular responses and also inform which candidate(s) will be most effective in blocking SARS-CoV-2 infection. We have streamlined the CRISPR engineering such that the proposed T4 vaccines can be constructed in about 4 weeks and the animal testing can be completed in about 12 weeks. The candidate vaccines will then be available for clinical trials and vaccine manufacture. The T4 vaccine will be exceedingly easy to manufacture, scale, and distribute globally, and could potentially lead to a breakthrough to avert the coronavirus crisis.

项目概要该提案旨在快速生成针对 2019 年新型冠状病毒 SARS-CoV-的候选疫苗 2. 自大约三个月前出现以来，该病毒已造成超过 120,000 例感染和 4,300 例感染全球范围内死亡，并迅速蔓延到包括美国在内的几乎每个国家。这个全球必须通过迅速制定医疗对策来立即应对卫生紧急情况。我们的噬菌体（噬菌体）T4 疫苗平台非常适合应对这一威胁。在 PI 中开发实验室证明，T4 疫苗可产生强大的体液和细胞免疫反应并在多种动物模型（包括小鼠、大鼠、兔子和猕猴。 T4 疫苗不需要佐剂，因为其表面结构模仿病原体病毒病原体的相关分子模式 (PAMP) 并刺激强大的先天性和适应性免疫。 120 x 86 nm 噬菌体 T4 衣壳包装有 171 kb 基因组，并装饰有两个非必需基因外衣壳蛋白； 870 个 Soc 分子（小外衣壳蛋白）和 155 个 Hoc 拷贝（高抗原性）外衣壳蛋白）。在具体目标1中，将通过整合SARS-冠状病毒构建一系列T4-冠状噬菌体通过 CRISPR 工程，单独或组合地形成 CoV-2 病毒体成分。编码基因整个刺突胞外域将在强 CAG 启动子的控制下插入噬菌体基因组。之上免疫，宿主细胞（免疫部位的肌细胞和抗原呈递细胞）摄取噬菌体颗粒并持续分泌胞外域三聚体，刺激免疫系统数周至数月。将插入S蛋白的受体结合域（RBD）的基因，使得RBD将被表达第870章每个衣壳的拷贝数。 E 蛋白的胞外域将与 Hoc 融合，并在每个衣壳中展示多达 155 个拷贝。最后，约 400 个 N 蛋白拷贝将被包装在衣壳内，作为支架核心的一部分。在具体目标 2 中，将评估上述 T4-corona 重组噬菌体是否诱导 SARS-CoV- 2 小鼠模型中病毒颗粒特异性免疫反应。将用纯化的噬菌体颗粒免疫小鼠肌肉注射，免疫反应将通过 ELISA、竞争性受体结合、ELISpot、和病毒中和试验。我们预计 T4 冠状病毒疫苗将产生强大的抗体和细胞反应，并告知哪些候选药物能够最有效地阻止 SARS-CoV-2 感染。我们简化了 CRISPR 工程，以便可以在约4周，动物试验约12周即可完成。候选疫苗将被可用于临床试验和疫苗生产。 T4 疫苗的制造将非常容易，规模化，并在全球范围内分布，并有可能在避免冠状病毒危机方面取得突破。