Headspace Metabolomics for Non-invasive Biomarker Discovery

先进的质谱学和数据处理工具的最新发展导致了护理点检测(POCT)的有效设计的蓬勃发展。方法是基于新陈代谢组学研究和创新传感器技术的新疾病生物标记物的发现。呼气分析是POCT的理想选择，因为它是非侵入性的；然而，它远远落后于血液测试。然而，各种疾病，如肺癌和喉癌、帕金森氏病和肺部疾病，已被证明具有特定的挥发性生物标记物“指纹”，到目前为止，这些标记物一直未得到充分利用。因此，我们建议将代谢组学转移到顶端空间，并研究微生物群落的挥发物，以探索尚未开发的诊断和筛查机会。该项目需要开发一种新的管道，用于在临床相关场景中发现呼吸挥发物(挥发性代谢物)中的生物标记物。针对口腔/肠道微生物群(生活在粘膜和口腔之间需氧/厌氧界面的复杂细菌群落)，影响口腔健康(龋齿形成、牙周炎、牙周炎)以及全身和心脏代谢健康，我们提出了一种独特的从实验台到临床实践的过渡方法。在该项目中，将使用典型挥发性代谢物的标准混合物来开发非靶向挥发性组学的分析方法。这将包括采样、GC-MS分析和数据处理方法。利用Gordon Ramage教授的实验室建立的现有的体外多物种生物膜模型，用不同的处理(如碳和氮的有效性)运行间歇生物膜发酵微观世界(体外)。管道的统计分析工具将从Gauchote-Lindsay小组的现有工具进行调整，以适应这些样本的挥发组，并将确定与治疗变化相关的生物标记物。最后，在健康的人类志愿者和患有牙周炎和肥胖相关内毒素血症的志愿者组中，将使用相同的分析管道在干预之前、期间和之后使用相同的分析管道来进行挥发性组特征描述，使用饮食或药物方法调节生物膜(体内)。在这里，将识别最终可能用于护理点传感器的生物标记物。目的和目标：该项目将回答这样一个问题：我们能否使用基于非侵入性呼吸采样的新的代谢组学方法，识别口腔健康的挥发性生物标记物作为通向全身健康的门户？其目的是制作一份可用于低成本护理点传感器或芯片实验室的化合物(最佳指纹)的入围名单。研究方法的新奇之处：代谢组学研究大多在血液、尿液或粪便等生物样本中进行。越来越多的证据表明，微生物活动调节人体内的广泛功能，相关疾病患者呈现出与健康受试者不同的微生物群。反过来，最近的研究还表明，微生物群落上方顶端空间的化学成分代表了新陈代谢和分解代谢的启动途径。因此，我们建议将代谢组学转移到顶端空间，并研究微生物群落的挥发组，以探索尚未开发的诊断和筛查机会。

Recent developments in advanced mass spectrometry and data treatment tools have led to a boom in the efficient design of point-of-care testing (POCT). Approaches are based on novel disease biomarkers discovery with metabolomics studies and innovative sensor technologies. Breath analysis is an ideal candidate for POCT because of its non-intrusive nature; it is, however, much less developed than blood testing. Nevertheless, diseases as varied as lung and throat cancers, Parkinson's disease and pulmonary diseases have been shown to possess specific volatile biomarkers "fingerprint" which have been, to date, under-utilised. We propose therefore to move metabolomics to the headspace and study the volatilome of microbial communities to explore yet untapped opportunities for diagnostic and screening.This project entails the development of a novel pipeline for biomarker discovery in the breath volatilome (volatile metabolites), in clinically-relevant scenarios. Focusing on the oral/gut microbiome (a complex community of bacteria living at the aerobic/anaerobic interface between mucosa and oral cavity), which influences oral health (cariogenesis, periodontitis, gingivitis) and systemic and cardio-metabolic health, we propose a unique transitional approach from bench to clinical practice.During the project, analytical methods will be developed for non-targeted volatilomics using standard mixtures of representative volatile metabolites. This will include sampling, GC-MS analysis and data processing methods. Batch biofilm fermentation microcosms (in vitro) with then be ran with various treatments (such as carbon and nitrogen availability), making use of existing in vitro multi-species biofilm models established in Prof Gordon Ramage's lab. The statistical analysis tool of the pipeline will be adapted from existing tools in the Gauchotte-Lindsay group to the volatilome of these samples and biomarkers that correlate with the variations in treatment will be identified. Finally, volatilome characterisation will be carried out using the same analytical pipelines in groups of healthy human volunteers and volunteers with periodontitis and obesity-associated endotoxaemia before, during and after an intervention using dietary or pharmacological approaches to modulate the biofilm (in vivo). Here biomarkers that could eventually be employed in point-of-care sensors will be identified.Aims and objectives:This project will answer the question: Can we identify volatile biomarkers for oral health as a portal to systemic health using a novel metabolomics approach based on non-intrusive breath sampling? The aim is to produce a shortlist of chemical compounds (optimal fingerprint) that could be employed in a low-cost point-of-care sensors or lab-on-a-chip.Novelty of the research methodology:Metabolomics studies have mostly been carried out in biological samples such as blood, urine or faeces. There is growing evidence that microbial activities regulate a broad range of functions in the human body and that patients with related illness(es) present different microbiome than healthy subjects. In turns, recent research also demonstrates that the chemical composition of the headspace above a microbial community is representative of the turned-on metabolic and catabolic pathways. We propose therefore to move metabolomics to the headspace and study the volatilome of microbial communities to explore yet untapped opportunities for diagnostic and screening.