Issitt Theo, Sweeney Sean T, Brackenbury William J, Redeker Kelly R
Department of Biology, University of York, York YO10 5DD, UK.
York Biomedical Research Institute, University of York, York YO10 5DD, UK.
Metabolites. 2022 Jun 27;12(7):599. doi: 10.3390/metabo12070599.
Volatile compounds, abundant in breath, can be used to accurately diagnose and monitor a range of medical conditions. This offers a noninvasive, low-cost approach with screening applications; however, the uptake of this diagnostic approach has been limited by conflicting published outcomes. Most published reports rely on large scale screening of the public, at single time points and without reference to ambient air. Here, we present a novel approach to volatile sampling from cellular headspace and mouse breath that incorporates multi-time-point analysis and ambient air subtraction revealing compound flux as an effective proxy of active metabolism. This approach to investigating breath volatiles offers a new avenue for disease biomarker discovery and diagnosis. Using gas chromatography mass spectrometry (GC/MS), we focus on low molecular weight, metabolic substrate/by-product compounds and demonstrate that this noninvasive technique is sensitive (reproducible at ~1 µg cellular protein, or ~500,000 cells) and capable of precisely determining cell type, status and treatment. Isolated cellular models represent components of larger mammalian systems, and we show that stress- and pathology-indicative compounds are detectable in mice, supporting further investigation using this methodology as a tool to identify volatile targets in human patients.
呼出气体中富含挥发性化合物，可用于准确诊断和监测一系列医疗状况。这提供了一种具有筛查应用的非侵入性、低成本方法；然而，这种诊断方法的采用受到已发表的相互矛盾结果的限制。大多数已发表的报告依赖于在单一时间点对公众进行大规模筛查，且未参考环境空气。在此，我们提出一种从细胞顶空和小鼠呼出气体中采集挥发性物质的新方法，该方法结合了多时间点分析和环境空气扣除，揭示化合物通量作为活跃代谢的有效指标。这种研究呼出气体挥发性物质的方法为疾病生物标志物的发现和诊断提供了一条新途径。使用气相色谱 - 质谱联用仪（GC/MS），我们聚焦于低分子量的代谢底物/副产物化合物，并证明这种非侵入性技术灵敏（在约1微克细胞蛋白或约500,000个细胞时可重复），且能够精确确定细胞类型、状态和治疗情况。分离的细胞模型代表了更大哺乳动物系统的组成部分，我们表明在小鼠中可检测到指示应激和病理的化合物，支持将该方法作为一种工具进一步研究，以识别人类患者中的挥发性靶点。
