Hyperpolarized Micro-NMR Platform for Sensitive Analysis of In Vitro Metabolic Flux in Living Cells.
Hyperpolarized Micro-NMR Platform for Sensitive Analysis of In Vitro Metabolic Flux in Living Cells.
复制标题
DOI：
10.1007/978-1-0716-1803-5_29
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
代谢代表细胞生化反应的整体，因此代谢分析可以揭示细胞的状态。例如，响应外部线索（例如药物治疗）的代谢变化可能是快速的，并且可能是治疗反应的早期指标。不幸的是，研究新陈代谢的传统技术，例如光学显微镜或质谱，在特异性和灵敏度方面存在功能限制。为了满足这一技术需求，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，能够快速无损地量化少量细胞（低至 10,000 个细胞）中的多种代谢通量。这种分析能力是通过核磁共振检测线圈的小型化以及内源代谢物的超极化来实现的。使用该工具，我们能够在 2 分钟内无损地量化癌症干细胞中丙酮酸到乳酸的通量，这是其他技术无法实现的。通过进一步优化，我们设想这种新颖的设备可以成为一个强大的分析平台，用于对质量限制样品中的代谢进行灵敏分析。
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.