Development of a novel and broadly applicable thermostable bacteriophage VLPs platforms for vaccine design, drug delivery, and imaging

开发新型且广泛适用的热稳定噬菌体 VLP 平台，用于疫苗设计、药物输送和成像

基本信息

批准号：
10282622
负责人：
Ebenezer Tumban
金额：
$ 38.93万
依托单位：
TEXAS TECH 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：
2020-02-24 至 2024-01-31
项目状态：
已结题

项目摘要

PROJECT SUMMARY Virus-like particles (VLPs) resemble - in size, structure, and immunogenicity - the virus from which the coat or envelope protein(s) are derived from except for the fact they lack a viral genome; VLPs are non-infectious and are safe. These features have been exploited to develop VLP-based vaccines against human papillomaviruses and hepatitis B virus; furthermore, coat proteins from ~70 viruses are currently being explored to develop VLP- based vaccines against these viruses. VLPs from some of these viruses have also been used as display platforms to develop chimeric VLPs displaying heterologous peptides from other infectious agents, tumor- associated antigens and other metabolic diseases. The goal of these chimeric VLPs is to induce antibodies against the heterologous antigen displayed on the platforms and not the platforms. In addition to serving as display platforms, VLPs have also been used for targeted delivery of drugs or cargo to specific cancer cells. While the candidate VLPs-based vaccines displaying heterologous peptides are very effective in animal studies, in the majority of studies, VLPs platforms (including adenoviral VLPs or dodecahedron) are derived from viruses that infect humans and in some cases, studies used VLPs platforms that had previously been used to immunize the general population; a good example is HBV vaccine, with a global infant vaccination coverage of 84% in 2015. Vaccines based on some of these platforms, with pre-existing antibodies in the general population, are likely to be less immunogenic in humans. Additionally, there is a limitation on the size of heterologous antigens that can be genetically displayed on some VLPs platforms making it challenging to display a single peptide with multiple epitopes on the same VLPs. Moreover, most of the VLPs platforms are temperature-sensitive making them less suitable in developing countries with poor refrigeration facilities. In this proposal, the PI will develop and characterize novel thermostable bacteriophage VLPs platforms using coat proteins from thermophilic viruses P23-77 and ΦIN93. P23-77 and ΦIN93 was isolated from bacteria that grow at 70-75 °C. Thus VLPs derived from these viruses are likely to be stable at room temperature (RT) or above RT. Additional benefit of these VLPs platforms is that because these viruses do not infect humans, the human population lacks pre-existing neutralizing antibodies against the VLPs platforms; thus, the immunogenicity of the platforms cannot be compromised by pre-existing antibodies. Also, many surface- exposed loops on the capsid may tolerate larger insertions of heterologous antigens. The PI will co-express three coat proteins from P23-77 and two coat proteins from ΦIN93. The PI will assess whether the coat proteins can assemble into VLPs, if they VLPs are thermostable, can tolerate heterologous peptide insertions from human papillomaviruses, and if VLPs are immunogenic in comparison to the virus(es).

项目概要病毒样颗粒 (VLP) 在大小、结构和免疫原性方面类似于病毒，其外壳或包膜蛋白的来源除了缺乏病毒基因组外，VLP 不具有传染性；这些特征已被用来开发基于 VLP 的人类乳头瘤病毒疫苗。此外，目前正在探索约 70 种病毒的外壳蛋白来开发 VLP- 来自其中一些病毒的 VLP 疫苗也已被用作展示。开发嵌合 VLP 的平台，展示来自其他感染因子、肿瘤的异源肽这些嵌合VLP的目标是诱导抗体。对抗平台上展示的异源抗原，而不是平台。展示平台上，VLP 也被用于将药物或货物靶向输送到特定的癌细胞。虽然展示异源肽的基于 VLP 的候选疫苗在动物身上非常有效研究中，大多数研究中，VLPs平台（包括腺病毒VLPs或十二面体）均衍生自从感染人类的病毒中分离出来，在某些情况下，研究使用了之前已被证实的 VLP 平台。用于对普通人群进行免疫；乙肝疫苗就是一个很好的例子，全球婴儿都接种疫苗 2015 年覆盖率达到 84%。基于其中一些平台的疫苗，其中含有预先存在的抗体一般人群中，对人类的免疫原性可能较低。此外，其大小也有限制。可以在某些 VLP 平台上进行基因展示的异源抗原，这使得在相同的 VLP 上展示具有多个表位的单个肽此外，大多数 VLP 平台都是如此。对温度敏感，因此不太适合制冷设施较差的发展中国家。在本提案中，PI 将使用以下方法开发和表征新型热稳定噬菌体 VLP 平台嗜热病毒 P23-77 和 ΦIN93 的外壳蛋白 P23-77 和 ΦIN93 是从细菌中分离出来的。在 70-75 °C 下生长，因此源自这些病毒的 VLP 在室温 (RT) 或这些 VLP 平台的另一个好处是，因为这些病毒不会感染人类，人群缺乏针对 VLP 平台的预先存在的中和抗体；此外，许多表面抗体不会损害平台的免疫原性。衣壳上暴露的环可以耐受更大的异源抗原插入。来自 P23-77 的三个外壳蛋白和来自 ΦIN93 的两个外壳蛋白 PI 将评估外壳是否存在。蛋白质可以组装成 VLP，如果它们是热稳定的，可以耐受异源肽插入来自人乳头瘤病毒，以及 VLP 与病毒相比是否具有免疫原性。