Development of intracellular VNARs as novel tools to dissect intracellular biological processes

开发细胞内 VNAR 作为剖析细胞内生物过程的新工具

• 批准号: BB/R009112/1
• 负责人: Christopher Scott
• 金额: $ 44.16万
• 依托单位: Queen's University Belfast
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR009112%2F1
• 关键词: Development; intracellular; VNARs; novel; tools

The human body has over 20,000 genes, from which the proteins that make up each human cell are generated. The study of the roles of these proteins is therefore central to our understanding of life. The best way to determine the role of individual proteins is to block their function. As Scientists, we have a number of approaches for inhibiting protein functions, including synthetic compounds prepared by Chemists that bind to the proteins as well as a number of genetic approaches that delete or deplete the gene that generates a particular protein. However, useful as these approaches are, they are not without significant drawbacks. For example, synthetic compounds normally only work on proteins that have particular types of docking sites (e.g. deep pockets or grooves in their surfaces), and it is very difficult (and expensive) to develop compounds that selectively target specific proteins using this approach. Approaches that target a particular protein's gene can be more selective, but often the cell adapts to the loss of a protein in the time it takes to generate experimental models in which the gene is deleted or depleted; this can limit the usefulness of the information that can be gleaned from such studies.In this LINK proposal, we aim to bring together academic biological expertise at Queen's University Belfast (QUB) with the industrial expertise of Elasmogen Ltd to develop a completely new approach that will generate a novel type of molecule that can rapidly and highly specifically bind to and inhibit target proteins and thereby enable the function of those proteins to be accurately determined without the potentially confounding effects of the approaches described above. VNARs are antibodies produced by the immune system of sharks, and having evolved 420 million years ago, they are the oldest antibodies so far identified in vertebrates, and they are also the smallest. Despite their small size, VNARs have a unique structure that gives them a potentially greater number of ways of binding to target proteins than the antibodies produced by our own immune systems; this means that VNARs exhibit an extremely high degree of specificity and selectivity for their target proteins.Elasmogen have established a unique set of over 100 billion individual synthetic VNARs, each of which has a different structure and can therefore bind to a different protein. Given the sheer number of VNARs in the Elasmogen collection, it will be possible to identify individual VNARs that can bind to each protein in the human body. Elasmogen and others have already demonstrated the ability of VNARs to bind to proteins expressed on the surface of human cells and proteins secreted from cells. In this project QUB and Elasmogen will collaborate together to examine the possibility of adapting VNARs so that they can get inside cells and target intracellular proteins. If successful, would open up a number of hugely exciting possibilities, including:1. Establishment of a novel class of highly selective, potent intracellular inhibitors that could become powerful research tools for dissecting important biological processes.2. Evidence that intracellular VNARs could potentially be used as novel therapeutics in a range of diseases.

人体有超过20，000个基因，构成每个人体细胞的蛋白质都是由这些基因产生的。因此，研究这些蛋白质的作用对我们理解生命至关重要。确定单个蛋白质作用的最佳方法是阻断它们的功能。作为科学家，我们有许多抑制蛋白质功能的方法，包括化学家制备的与蛋白质结合的合成化合物，以及一些删除或耗尽产生特定蛋白质的基因的遗传方法。然而，尽管这些方法是有用的，但它们并非没有显著的缺点。例如，合成化合物通常只作用于具有特定类型的对接位点（例如其表面的深口袋或凹槽）的蛋白质，并且使用这种方法开发选择性靶向特定蛋白质的化合物非常困难（并且昂贵）。靶向特定蛋白质基因的方法可能更具选择性，但通常细胞在生成基因缺失或耗尽的实验模型所需的时间内适应蛋白质的丢失;这可能限制了从这些研究中收集的信息的有用性。在这一链接提议中，我们的目标是汇集学术生物学专业知识在贝尔法斯特女王大学（QUB）与Elasmogen有限公司的工业专业知识，开发一种全新的方法，将产生一种新型的分子，可以快速，高度特异性地结合并抑制靶蛋白，从而能够准确地确定那些蛋白的功能，而没有上述方法的潜在混淆效应。VNAR是鲨鱼免疫系统产生的抗体，在4.2亿年前进化而来，是迄今为止在脊椎动物中发现的最古老的抗体，也是最小的。尽管VNARs的体积很小，但它们具有独特的结构，这使得它们与靶蛋白的结合方式可能比我们自身免疫系统产生的抗体更多;这意味着VNAR对其靶蛋白表现出极高的特异性和选择性。Elasmogen已经建立了一套独特的超过1000亿个单独的合成VNAR，每一种都具有不同的结构，因此可以结合不同的蛋白质。鉴于Elasmogen集合中VNAR的绝对数量，将有可能鉴定出可以与人体中的每种蛋白质结合的单个VNAR。Elasmogen和其他人已经证明了VNAR与人细胞表面表达的蛋白质和细胞分泌的蛋白质结合的能力。在这个项目中，QUB和Elasmogen将合作研究调整VNAR的可能性，使它们能够进入细胞内并靶向细胞内蛋白质。如果成功的话，将开启一系列令人兴奋的可能性，包括：1。建立一类新型的高选择性、有效的细胞内抑制剂，可以成为解剖重要生物过程的有力研究工具。细胞内VNAR可能在一系列疾病中用作新型治疗剂的证据。

项目成果

FLIP(L): the pseudo-caspase.

• DOI: 10.1111/febs.15260
• 发表时间: 2020-10
• 期刊: The FEBS journal
• 作者: Smyth P;Sessler T;Scott CJ;Longley DB
• 通讯作者: Longley DB

Development of next generation nanomedicine-based approaches for the treatment of cancer: we've barely scratched the surface.

• DOI: 10.1042/bst20210343
• 发表时间: 2021-11-01
• 期刊: Biochemical Society transactions
• 影响因子: 3.9
• 作者: Tracey SR;Smyth P;Barelle CJ;Scott CJ
• 通讯作者: Scott CJ

Evaluation of variable new antigen receptors (vNARs) as a novel cathepsin S (CTSS) targeting strategy.

• DOI: 10.3389/fphar.2023.1296567
• 发表时间: 2023
• 期刊: Frontiers in pharmacology
• 影响因子: 5.6