WOrM Drug Models: Whole Organism Metabolomic Drug Models to improve holistic understanding of therapeutic performance

该项目旨在建立秀丽隐杆线虫作为模型系统，以确定药物对整个生物体代谢组学的全球变化的影响，以提高治疗效果。背景：代谢组学被认为是生物体生理状态和表型的功能性读数，可以通过药物治疗显着改变。人体代谢组学的有效建模具有挑战性。这是因为人类是大型复杂动物，尚未被完全理解，并且在科学和社会经济方面都具有挑战性。因此，国际上有一股强大的动力来取代、减少和改进动物在研究中的使用，以便将这一宝贵资源保留给最强的治疗候选药物，同时也提供一种机制来改进那些在早期开发阶段失败的治疗方法。考虑到这一点，有必要开发创新的解决方案或重新利用现有的模型，以提供对治疗效果的关键见解。一个潜在解决方案的极好例子是模式生物秀丽隐杆线虫。秀丽隐杆线虫，一种自由生活的线虫，是地球上最被完全理解的动物。这是由于其体积小（<1毫米），生成时间短（<3天），光学透明性，遗传变异的可用性以及不受内政部动物法规的限制。迄今为止，秀丽隐杆线虫的完整基因组、蛋白质组和连接组已经被绘制出来。然而，目前还没有现成的关于秀丽隐杆线虫的代谢信息。方法：该项目将利用药学院和诺丁汉大学世界领先的分析生物科学中心（CAB）和纳米与微尺度研究中心（nmRC）的分析设施，填补药物反应的全生物代谢组学的重要知识空白。CAB是高分辨率超高性能液相色谱-串联质谱（UHPLC-MS/MS）的所在地。UHPLC-MS/MS将用于破译被广泛使用的药物（阿替洛尔、对乙酰氨基酚、氟西汀、奥美拉唑和辛伐他汀）影响的线虫匀浆液的代谢组学特征。这将为具有已知药物活性的疗法提供校准的代谢组学结果。线虫也将面临下一代疗法的挑战，“反向前药”，它通过引入电活动来自我聚合，以阻止癌症的生长。校准药物样本的代谢组将为下一代疗法的生理状态提供重要的见解，并为提高性能提供途径。nmRC拥有世界上唯一的学术上的3D-OrbiSIMS，能够进行高空间和化学分辨率的三维质谱分析。LC-MS鉴定的关键代谢组学变化将与3D-OrbiSIMS在空间上协调。这一科学首次将用于制作秀丽隐杆线虫全生物体代谢组学的“谷歌街景”。这将为研究人员提供宝贵的资源，以提高他们对蠕虫生理状态的理解，并进一步了解秀丽隐杆线虫作为复杂哺乳动物生物化学模型的知识。影响：这一多学科项目将提高对治疗方法对代谢途径影响的认识，并将有助于生产新的和改进的药物，以预防和治疗疾病，如癌症。它还将为博士候选人提供多样化的培训机会。在经验丰富的监督团队的支持下，所进行的研究将为建立新的和改进的药物输送模型和分析铺平道路。

CDT theme alignment: Predictive Pharmaceutical Sciences, Pharmaceutical Process Engineering and Complex Product Characterisation This project will aim to establish Caenorhabditis elegans as a model system to determine the influence of pharmaceuticals on global shifts in Whole Organism Metabolomics, to improve therapeutic performance. BACKGROUND: Metabolomics is considered as the functional readout of the physiological state and phenotype of an organism, which can be significantly transformed by drug treatment. Effective modelling of metabolomics in humans is challenging. This is because humans are large complex animals, which are not completely understood and can be both scientifically and socio-economically challenging to characterise. Therefore, there is a strong international drive to replace, reduce and refine the use of animals in research so that this precious resource is reserved for the strongest therapeutic candidates, whilst also providing a mechanism to improve those therapeutics that fail at early-stage development. Bearing this in mind there is a need to develop innovative solutions or repurpose existing models that can provide key insights into therapeutic performance. An excellent example of a potential solution is the model organism C. elegans. C. elegans, a free-living nematode, is the most completely understood animal on the planet. This is due to its small size (<1 mm), short generation time (<3 days), optical transparency, availability of genetic variants and exclusion from Home Office animal regulations. To date, the complete genome, proteome and connectome for C. elegans have been mapped. However, currently there is no readily available metabolic information on C. elegans. METHOD: This project will fill the important knowledge gaps in whole organism metabolomics in response to drugs by harnessing the School of Pharmacy's & University of Nottingham's world leading analytical facilities of the Centre for Analytical Bioscience (CAB) and Nanoscale & Microscale Research Centre (nmRC). The CAB is home to a high-resolution ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). UHPLC-MS/MS will be used to decipher metabolomic profiles of nematode homogenates challenged to widely used pharmaceuticals, with well-established literature on influence on physiological state (atenolol, acetaminophen, fluoxetine, omeprazole and simvastatin). This will provide a calibrated metabolomic outcome to therapeutics with known pharmaceutical activity. Nematodes will also be challenged to next generation therapeutics, "reverse prodrugs," which self-polymerise through the introduction of electrical activity to stop cancer growth. The metabolome of the calibrated drug samples will provide an important insight to the physiological state of next-gen therapies and provide pathways to improve performance. The nmRC hosts the world's only academically situated 3D-OrbiSIMS, capable of high spatial and chemical resolution three-dimensional mass spectrometry. The key metabolomic shifts identified from LC-MS will be spatially coordinated with 3D-OrbiSIMS. This scientific first will be used to produce a "Google street view" of Whole Organism Metabolomics for C. elegans. This will be a valuable resource for researchers to improve their understanding of the physiological state of WOrMs and those furthering the knowledge of C. elegans as a model for complex mammalian biochemistry. IMPACT: This multidisciplinary project will improve the understanding of the influence of therapeutics on metabolic pathways and will contribute to the production of new and improved drugs to prevent and treat diseases, such as cancer. It will also provide diverse training opportunities for the PhD candidate. With support form the experienced supervisory team the research conducted will pave-the-way towards establishing new and improved models and analytics for drug delivery.