Empty virus-like particles (eVLPs) as bio-compatible targeted drug-delivery vehicles

空病毒样颗粒（eVLP）作为生物相容性靶向药物递送载体

• 批准号: BB/I002294/1
• 负责人: George Lomonossoff
• 金额: $ 45.98万
• 依托单位: John Innes Centre
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2011
• 资助国家: 英国
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FI002294%2F1
• 关键词: Empty; virus; like; particles; eVLPs

A major challenge in pharmacology is to devise methods whereby drugs can be delivered specifically to target tissues. This is a particular issue in the case of anti-cancer drugs which usually discriminate between cancerous and normal cells by the fact that the cancer cells are dividing more rapidly. However anti-cancer drugs are toxic to all cells and thus often have severe side-effects. To avoid this, it would clearly be desirable to target the drug molecule specifically to the cancerous tissue. A potential means of achieving this would be to package or encapsulate the drug molecules inside a particle which is designed to bind solely to the cancerous tissue. Such encapsulation would have the additional advantage of protecting the drug from breakdown in blood plasma. For this to become a reality it will be necessary to develop particles which can be modified on their outer surface to achieve the desired targeting and which can contain drug molecules. The particles need to be small enough to be able to move in the bloodstream, are not toxic and to be able to enter cells. One type of particle which has all these characteristics is the plant virus, cowpea mosaic virus (CPMV), which can be produced in large quantities by infecting plants. Previous research has shown that it is possible to 'stick' molecules on the surface of CPMV particles which enable them to be targeted to specific cells. Despite these advantages, CPMV particles have not, to date, been exploited as a drug-delivery vehicle. The reason for this is that particles produced by the infection of plants are already filled with the virus' own genetic material. This means that there is little or no room to put anything else, such as drug molecules, inside the particles. Even if such molecules could somehow be squeezed in, there would still be concerns about administering particles containing viral genetic material, even though it is from a plant virus that cannot infect animals. This project addresses the issue of the genetic material within the virus particles by exploiting the recent discovery at the John Innes Centre of a method of producing large quantities of pure empty (lacking genetic material) virus-like particles (eVLPs) of CPMV in plants. The method involves simultaneously expressing genes coding for a precursor of the viral coat proteins and the enzyme used to process it using a recently developed highly efficient plant transient expression system. Applying this approach we will produce particles which are modified so that they will specifically bind to proteins expressed on the surface of cancerous, but not normal, cells. This modification will be done either chemically or by making genetic fusions to the virus coat protein. We will investigate the best way of loading the targeted particles with the anti-cancer drug, gemcitabine. To do this we will make use of pores in the virus particles which allow small molecules to enter under certain conditions. The ability of the targeted particles to deliver the drug to cancer cells will be investigated by testing their ability to bind to and kill cancers in culture. This will be the first important step in potentially developing brand new therapeutic agents to tackle human cancer.

药理学的一个主要挑战是设计可以将药物特异性递送至靶组织的方法。对于抗癌药物而言，这是一个特殊的问题，抗癌药物通常通过癌细胞分裂更快的事实来区分癌细胞和正常细胞。然而，抗癌药物对所有细胞都有毒，因此常常具有严重的副作用。为了避免这种情况，显然需要将药物分子特异性地靶向癌组织。实现这一目标的一种潜在方法是将药物分子包装或封装在颗粒内，该颗粒被设计为仅与癌组织结合。这种封装具有保护药物免于在血浆中分解的额外优点。为了使这一目标成为现实，有必要开发可以对其外表面进行修饰以实现所需靶向并且可以包含药物分子的颗粒。这些颗粒必须足够小，以便能够在血液中移动、无毒并且能够进入细胞。具有所有这些特征的一类颗粒是植物病毒，即豇豆花叶病毒（CPMV），它可以通过感染植物而大量产生。先前的研究表明，可以将分子“粘附”在 CPMV 颗粒的表面，从而使它们能够靶向特定细胞。尽管有这些优点，CPMV 颗粒迄今为止尚未被用作药物输送载体。原因是植物感染后产生的颗粒中已经充满了病毒自身的遗传物质。这意味着颗粒内部几乎没有空间可以放置其他任何东西，例如药物分子。即使这些分子可以以某种方式被挤进去，人们仍然会担心施用含有病毒遗传物质的颗粒，即使它来自不能感染动物的植物病毒。该项目通过利用约翰·英尼斯中心最近发现的一种在植物中生产大量纯空（缺乏遗传物质）CPMV 病毒样颗粒（eVLP）的方法来解决病毒颗粒内遗传物质的问题。该方法涉及使用最近开发的高效植物瞬时表达系统同时表达编码病毒外壳蛋白前体的基因和用于加工它的酶。应用这种方法，我们将生产经过修饰的颗粒，以便它们能够特异性地结合癌性细胞而非正常细胞表面表达的蛋白质。这种修饰可以通过化学方法或通过与病毒外壳蛋白进行基因融合来完成。我们将研究用抗癌药物吉西他滨负载靶向颗粒的最佳方法。为此，我们将利用病毒颗粒中的孔，这些孔允许小分子在某些条件下进入。将通过测试靶向颗粒与培养物中的癌症结合并杀死癌症的能力来研究靶向颗粒将药物递送至癌细胞的能力。这将是开发治疗人类癌症的全新治疗药物的第一个重要步骤。

项目成果

Applications of plant viruses in bionanotechnology.

• DOI: 10.1007/82_2011_184
• 发表时间: 2014
• 期刊: Current topics in microbiology and immunology
• 作者: Lomonossoff GP;Evans DJ
• 通讯作者: Evans DJ

The structures of a naturally empty cowpea mosaic virus particle and its genome-containing counterpart by cryo-electron microscopy.

• DOI: 10.1038/s41598-017-00533-w
• 发表时间: 2017-04-03
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Hesketh EL;Meshcheriakova Y;Thompson RF;Lomonossoff GP;Ranson NA
• 通讯作者: Ranson NA

Stability of plant virus-based nanocarriers in gastrointestinal fluids.

• DOI: 10.1039/c7nr07182e
• 发表时间: 2018-01-25
• 期刊: Nanoscale
• 影响因子: 6.7
• 作者: Berardi A;Evans DJ;Baldelli Bombelli F;Lomonossoff GP
• 通讯作者: Lomonossoff GP

Virus Hybrids as Nanomaterials: Methods and Protocols

作为纳米材料的病毒杂交体：方法和方案

• 发表时间: 2013
• 作者: Lin, Baochuan;Ratna, Banahalli
• 通讯作者: Ratna, Banahalli