mRNA COVID-19 Vaccines Delivered with Plant Virus/Polymer Devices

通过植物病毒/聚合物设备提供的 mRNA COVID-19 疫苗

• 批准号: 10231938
• 负责人: Jonathan Kyle Pokorski
• 金额: $ 23.7万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10231938
• 关键词: 2019-nCoV; Adjuvant; Animals; Antibody titer measurement; Antigens; Area; COVID-19; COVID-19 pandemic; COVID-19 testing; COVID-19 vaccine; Cells; Civilization; Cold Chains; Development; Devices; Disease Outbreaks; Dose; Equipment; Evaluation; Formulation; Foundations; Future; Genetic Code; Goals; Health; Human; Immune; Immune system; Immunity; Implant; In Situ; In Vitro; Industrialization; Innate Immune System; Livestock; Messenger RNA; Modeling; Moderna COVID-19 vaccine; Mus; Nanotechnology; National Institute of Allergy and Infectious Disease; Nucleic Acid Vaccines; Phase I Clinical Trials; Plant Viruses; Plants; Polymers; Population; Process; Production; Proteins; RNA; RNA Interference; RNA vaccine; Resources; SARS-CoV-2 spike protein; Scheme; Signal Transduction; Speed; Technology; Tertiary Protein Structure; Time; Tobacco Mosaic Virus; Vaccination; Vaccines; Viral Proteins; Virus; Virus Diseases; Virus-like particle; Vision; Zoonoses; base; biosafety level 3 facility; in vivo; lipid nanoparticle; mRNA delivery; melting; mouse model; nanocarrier; nanoparticle; nucleic acid delivery; pandemic disease; pathogen; prevent; protein expression; receptor; reconstitution; response; scale up; tissue culture; tool; vaccine candidate; vaccine delivery; vaccine distribution; vaccine evaluation; vaccine trial

Summary The rapid development, distribution and administration of a COVID-19 vaccine to the global population is the most effective approach to quell this pandemic and future reoccurrence. Vaccination must occur broadly both in well-resourced and resource-poor areas of the world to be most effective. We propose a vaccine delivery device based on a plant virus/polymer blend. The manufacturing scheme proposed would allow for timely production at scale; most importantly, the vaccine delivery device would be stable outside of the cold- chain and only require a single-dose administration. We propose to package an mRNA vaccine encoding the SARS-CoV-2 S protein (and domains thereof) in virus-like particles (VLPs) derived from the plant virus Tobacco mosaic virus (TMV). TMV can be reconstituted in vitro or in plants to carry heterologous RNA and its utility in mRNA vaccine delivery has been demonstrated. Plant VLPs are highly visible to the immune system, these vaccine delivery agents boost immunity through signaling via multiple receptors of the innate immune system; therefore, the VLP serves as carrier and well-defined adjuvant. Furthermore, TMV offers high thermal stability – therefore overcoming cold chain requirements; but more importantly the mRNA-laden TMV vaccine candidates offer such a high degree of stability that they can withstand the rigors of melt-processed device manufacture, therefore allowing the formulation of slow-release implants or microneedle patches for single-administration. We will produce mRNA-laden TMV and then apply melt-processing tools manufacture the slow-release implants. We have developed a microextruder to manufacture protein/polymer blends that allows for the formulation of plant virus-based vaccines into slow-release devices. We have already demonstrated that plant virus-based vaccines can withstand the rigor of melt-processing. Under the tenure of this R21, we will package SARS-CoV-2 derived mRNA cassettes into TMV and verify mRNA delivery and protein expression in immune cells (Aim 1). We will formulate slow-release VLP/polymer (PLGA) blends and determine VLP (mRNA-laden TMV) release rates from the VLP/polymer blends (Aim 2). We will determine antibody titers and cellular anti-SARS-CoV-2 responses of the mRNA-laden TMV vaccine candidates in mice and compare soluble (prime-boost) vs. implant vaccine candidates (single administration). We will determine whether vaccination yields neutralizing SARS-CoV-2 sera and cellular responses; in vitro and in vivo SARD-CoV-2 challenge studies will be performed (Aim 3). Our technology brings unique attributes in that it does not require the cold chain for distribution and would enable vaccination upon single administration. Single- administration vaccines could also enable vaccination of livestock and this could be a step forward to meet the goals of the One Health Initiative to prevent future outbreaks.

概括 COVID-19 疫苗的快速开发、向全球人群的分发和管理 是平息这一流行病和未来再次发生的最有效方法。必须进行疫苗接种 广泛地在世界资源丰富和资源贫乏的地区发挥最有效的作用。我们提出疫苗 基于植物病毒/聚合物混合物的递送装置。拟议的制造方案将允许 及时规模化生产；最重要的是，疫苗输送装置在寒冷的环境下也能保持稳定 链并且只需要单剂量给药。我们建议包装一种 mRNA 疫苗，编码 源自植物的病毒样颗粒 (VLP) 中的 SARS-CoV-2 S 蛋白（及其结构域） 病毒 烟草花叶病毒（TMV）。 TMV可以在体外或在植物中重组以携带异源RNA 其在 mRNA 疫苗递送中的效用已得到证实。植物 VLP 对免疫系统高度可见 系统中，这些疫苗递送剂通过先天性多个受体发出信号来增强免疫力 免疫系统;因此，VLP 可作为载体和明确的佐剂。此外，TMV 还提供高 热稳定性——因此克服了冷链要求；但更重要的是携带 mRNA 的 TMV 候选疫苗具有高度的稳定性，可以承受熔融加工的严酷考验 设备制造，因此可以配制缓释植入物或微针贴片 单一管理。我们将生产载有 mRNA 的 TMV，然后应用熔融加工工具 制造缓释植入物。我们开发了一种微型挤出机来制造蛋白质/聚合物 混合物可以将基于植物病毒的疫苗配制到缓释装置中。我们已经 证明基于植物病毒的疫苗可以承受严格的熔化加工。任职期间 在此 R21 中，我们将把 SARS-CoV-2 衍生的 mRNA 盒包装到 TMV 中并验证 mRNA 递送和蛋白质 在免疫细胞中表达（目标 1）。我们将配制缓释 VLP/聚合物 (PLGA) 混合物并确定 VLP（载有 mRNA 的 TMV）从 VLP/聚合物混合物中的释放率（目标 2）。我们将测定抗体滴度 以及携带 mRNA 的 TMV 候选疫苗在小鼠体内的细胞抗 SARS-CoV-2 反应，并进行比较 可溶性（初免-加强）与植入候选疫苗（单次给药）。我们将确定是否 疫苗接种可中和 SARS-CoV-2 血清和细胞反应；体外和体内 SARD-CoV-2 将进行挑战研究（目标 3）。我们的技术带来了独特的属性，因为它不 需要冷链进行分配，并且可以一次性接种疫苗。单身的- 管理疫苗还可以对牲畜进行疫苗接种，这可能是实现这一目标的一个进步 “同一个健康倡议”的目标是预防未来的疫情爆发。