Developing drug discovery assays for primary human immune cells using high-throughput MALDI-TOF mass spectrometry

在药物发现管道的高通量筛选（HTS）过程中，体外试验用于评估化合物文库对特定药物靶点的功效。目前，荧光和化学发光是HTS检测中最常用的方法。作为一种令人兴奋的新工具，下一代基质辅助激光解吸/电离（MALDI）飞行时间（TOF）质谱（MS）仪器已用于药物发现的HTS方法。与目前使用的基于荧光的方法相比，它们具有高灵敏度、鲁棒性、快速和无标记的优点。此外，这些新的MALDI-TOF-MS仪器现在可以与全自动平台和工作流程集成。近年来，已经证明基于MALDI-TOF质谱的体外测定可用于直接监测各种酶促反应的产物和底物。这些直接读数已被用于筛选小分子激活剂和抑制剂对抗各种蛋白质靶标，并显示与基于荧光的分析结果相关。将该技术转移到大规模药物发现工作流程中，将为使用无标签底物提供巨大的优势。MALDI-TOF质谱也可用于细胞测定，尽管该技术在工业环境中尚未很好地建立，因此需要对细胞进行表征。为了提高从可能的药物化合物鉴定到临床的翻译，非常需要生物学相关的模型。这尤其适用于人类原代和诱导多能干细胞衍生的免疫细胞，它们已经用于早期药物发现，以更好地描述人类生物学。它们在自身免疫和癌症中发挥重要作用。目前，细胞分析是基于已建立的细胞系，因此使用人类原代和诱导多能干细胞衍生的免疫细胞分析将允许在药物发现管道中实施生物学相关模型，从而提高HTS过程的效率。该博士项目将专注于开发用于药物发现的生化高通量筛选工作流程，以及在MALDI-TOF MS平台上探索和进一步了解基于细胞的模型。这些技术将应用于免疫反应中的有吸引力的目标，以及原代免疫细胞，因为它们是抗炎药物和免疫肿瘤学药物的目标。最近，许多药物发现活动都集中在利用自身免疫系统对抗癌症的免疫疗法上。该项目由纽卡斯尔大学Matthias Trost教授的实验室进行，并将与葛兰素史克公司（GSK）合作进行。

In the high-throughput screening (HTS) process of the drug discovery pipeline, in vitro assays are used to evaluate the efficacy of compound libraries against a specific drug target. Currently, fluorescence and chemiluminescence are the most common methods used in HTS assays. As an exciting new tool, next generation matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) instruments have been implemented for HTS approaches in drug discovery. They provide the advantage of being highly sensitive, robust, fast and label-free in comparison to the currently used fluorescence-based methods. Moreover, these new MALDI-TOF-MS instruments can now be integrated with full automation platforms and workflows. In recent years, it has been demonstrated that MALDI-TOF MS based in vitro assays can be used to directly monitor products and substrates of a diverse range of enzymatic reactions. These direct readouts have since been used for screening small molecule activators and inhibitors against a variety of protein targets and shown to correlate with fluorescent based assay results. Transferring this technique onto large scale drug discovery workflows will provide the great advantage to work with label free substrates. MALDI-TOF MS can also be used for cellular assays, although this technique is not well established in the industrial setting and therefore requires characterisation of cells. There is a great need for biologically relevant models in order to improve the translation from possible drug compound identifications into the clinic. In particular, this applies to human primary and induced pluripotent stem cell derived immune cells, which are already used in early phase drug discovery, to better describe the human biology. They play important roles in auto-immunity and in cancer. Currently, cellular assays are based on established cell lines and therefore using human primary and induced pluripotent stem cell derived immune cells assays will allow the implementation of biologically relevant models in the drug discovery pipeline which can improve the efficiency of HTS processes.The PhD project will focus on the development of biochemical high-throughput screening workflows for drug discovery, as well as explore and further understand cell-based models on a MALDI-TOF MS platform. These techniques will be applied to attractive targets within the immune response, as well as primary immune cells as they are targets for anti-inflammatory drugs and drugs in immune oncology. Recently, many drug discovery campaigns are focusing on immunotherapy that uses the own immune system to fight cancer. This project is based in the laboratory of Professor Matthias Trost at Newcastle University and will be conducted in collaboration with GlaxoSmithKline (GSK).