Memorial Sloan Kettering Cancer Center, New York, NY, USA.
Methods Mol Biol. 2022;2393:561-569. doi: 10.1007/978-1-0716-1803-5_29.
Metabolism represents an ensemble of cellular biochemical reactions, and thus metabolic analyses can shed light on the state of cells. Metabolic changes in response to external cues, such as drug treatment, for example, can be rapid and potentially an early indicator of therapeutic response. Unfortunately, conventional techniques to study metabolism, such as optical microscopy or mass spectrometry, have functional limitations in specificity and sensitivity. To address this technical need, we developed a sensitive analytical tool based on nuclear magnetic resonance (NMR) technology, termed hyperpolarized micro-NMR, that enables rapid quantification of multiple metabolic fluxes in a small number of cells, down to 10,000 cells, nondestructively. This analytical capability was achieved by miniaturization of an NMR detection coil along with hyperpolarization of endogenous metabolites. Using this tool, we were able to quantify pyruvate-to-lactate flux in cancer stem cells nondestructively within 2 min, which has not been possible with other techniques. With further optimization, we envision that this novel device could be a powerful analytical platform for sensitive analysis of metabolism in mass-limited samples.
代谢代表了一组细胞内的生化反应，因此代谢分析可以揭示细胞的状态。细胞对外界刺激（如药物治疗）的代谢变化可能非常迅速，并且可能是治疗反应的早期指标。不幸的是，用于研究代谢的传统技术，如光学显微镜或质谱法，在特异性和灵敏度方面存在功能限制。为了解决这一技术需求，我们开发了一种基于核磁共振（NMR）技术的灵敏分析工具，称为超极化微 NMR，可在不破坏细胞的情况下，快速定量少量细胞（低至 10000 个细胞）中的多种代谢通量。这种分析能力是通过 NMR 检测线圈的小型化以及内源性代谢物的超极化来实现的。使用该工具，我们能够在 2 分钟内非破坏性地定量癌症干细胞中的丙酮酸到乳酸通量，这是其他技术无法实现的。通过进一步优化，我们设想这种新型设备可以成为一种强大的分析平台，用于对有限量样本中的代谢进行灵敏分析。
