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

• 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
• 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

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.

项目摘要 该提案旨在迅速针对2019年新型冠状病毒SARS-COV - 2.自三个月前出现以来，该病毒已引起超过120,000次感染和4,300次感染 全世界的死亡人数正在迅速传播到包括美国在内的每个国家。这个全球 必须通过快速发展内侧对策来立即解决健康紧急情况。 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的开发中开发 实验室，T4疫苗已被证明可以产生强大的体液和细胞免疫反应 并赋予多种动物模型中的炭疽和鼠疫的完全保护，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了病原体 病毒病原体的相关分子模式（PAMP），并刺激强烈的先天和适应性免疫。 120 x 86 nm噬菌体T4 Capsid用171 kb的基因组包装，并用两个非必需品装饰 外带状蛋白； 870个SOC分子（小型外皮蛋白）和155份HOC（高度抗原 外带状蛋白）。在特定的目标1中，将通过合并SARS-将构建一系列T4-Corona噬菌体 COV-2病毒成分单独和组合，通过CRISPR工程。编码的基因 在强CAG启动子的控制下，整个尖峰外生域将被插入噬菌体基因组中。之上 免疫，宿主细胞（在免疫部位的肌细胞和抗原呈递细胞）占据噬菌体 颗粒并连续分泌骨质域的三聚体，刺激免疫系统数周到几个月。 将插入S蛋白的受体结合结构域（RBD）的基因，以表达RBD 在宿主细胞以及大肠杆菌中作为SOC融合蛋白，然后将其显示在噬菌体帽上，最高为870 人均副本。 E蛋白质的外生域将融合成HOC，并显示高达155份的人均份。 最后，作为脚手架芯的一部分，将在衣壳内包装约400份N蛋白的副本。 在特定的目标2中，将评估上述T4-Corona重组噬菌体以启发SARS-COV- 2小鼠模型中的2个病毒体特异性免疫反应。小鼠将用纯化的噬菌体颗粒免疫 肌肉内和免疫反应将通过ELISA，竞争受体结合，ELISPOT， 和病毒中和测定法。我们预计T4-Corona疫苗将引起强大的抗体和细胞 反应，并告知哪些候选人最有效地阻止SARS-COV-2感染。 我们简化了CRISPR工程，以便可以在 大约4周，可以在大约12周内完成动物测试。候选疫苗将是 可用于临床试验和疫苗制造。 T4疫苗非常容易生产， 扩展并在全球范围内分发，并有可能导致突破性避免冠状病毒危机。