Department of Radiology, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA.
Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA.
Sci Adv. 2017 Jun 16;3(6):e1700341. doi: 10.1126/sciadv.1700341. eCollection 2017 Jun.
Metabolic reprogramming is widely considered a hallmark of cancer, and understanding metabolic dynamics described by the conversion rates or "fluxes" of metabolites can shed light onto biological processes of tumorigenesis and response to therapy. For real-time analysis of metabolic flux in intact cells or organisms, magnetic resonance (MR) spectroscopy and imaging methods have been developed in conjunction with hyperpolarization of nuclear spins. These approaches enable noninvasive monitoring of tumor progression and treatment efficacy and are being tested in multiple clinical trials. However, because of their limited sensitivity, these methods require a larger number of cells, on the order of 10, which is impractical for analyzing scant target cells or mass-limited samples. We present a new technology platform, a hyperpolarized micromagnetic resonance spectrometer (HMRS), that achieves real-time, 10-fold more sensitive metabolic analysis on live cells. This platform enables quantification of the metabolic flux in a wide range of cell types, including leukemia stem cells, without significant changes in viability, which allows downstream molecular analyses in tandem. It also enables rapid assessment of metabolic changes by a given drug, which may direct therapeutic choices in patients. We further advanced this platform for high-throughput analysis of hyperpolarized molecules by integrating a three-dimensionally printed microfluidic system. The HMRS platform holds promise as a sensitive method for studying metabolic dynamics in mass-limited samples, including primary cancer cells, providing novel therapeutic targets and an enhanced understanding of cellular metabolism.
代谢重编程被广泛认为是癌症的一个标志，了解代谢物转化率或“通量”所描述的代谢动态，可以揭示肿瘤发生和对治疗反应的生物学过程。为了实时分析完整细胞或生物体中的代谢通量，已经开发出了与核自旋极化相结合的磁共振（MR）光谱和成像方法。这些方法能够非侵入性地监测肿瘤进展和治疗效果，并且正在多项临床试验中进行测试。然而，由于其灵敏度有限，这些方法需要更多数量的细胞，大约为 10 个，这对于分析稀少的靶细胞或质量有限的样本是不切实际的。我们提出了一种新的技术平台，即超极化微磁共振光谱仪（HMRS），该平台可实时进行 10 倍更灵敏的活细胞代谢分析。该平台能够定量分析包括白血病干细胞在内的多种细胞类型的代谢通量，而对细胞活力没有显著影响，这允许与下游分子分析同时进行。它还可以快速评估给定药物引起的代谢变化，从而为患者的治疗选择提供指导。我们通过集成三维打印微流控系统进一步推进了这个用于高通量分析超极化分子的平台。HMRS 平台有望成为一种在质量受限的样本中研究代谢动态的灵敏方法，包括原发性癌细胞，为新的治疗靶点和对细胞代谢的更深入理解提供了可能。
