Univ. of Wisconsin-Madison, United States.
Morgridge Institute for Research, United States.
J Biomed Opt. 2020 Mar;25(3):1-13. doi: 10.1117/1.JBO.25.3.036502.
Glioblastoma multiforme (GBM) is the most frequently diagnosed adult primary brain malignancy with poor patient prognosis. GBM can recur despite aggressive treatment due to therapeutically resistant glioblastoma stem cells (GSCs) that may exhibit metabolic plasticity.
Intrinsic nicotinamide adenine dinucleotide (NADH) fluorescence can be acquired with fluorescence lifetime imaging microscopy (FLIM) to examine its bound and free metabolic states in GSC and GBM tissues.
We compared the mean NADH fluorescence lifetime in live human GSCs and normal neural stem cells and validated those results by measuring oxygen consumption rates (OCRs). We also examined the role that invasive versus less-invasive GSCs had on tumor metabolism by measuring the mean NADH lifetimes and the relative amount of the longer-lived component of NADH and correlated these results with survival in an orthotopic mouse xenograft model.
Mean NADH lifetime, amount of bound NADH, and OCR were increased in GSCs. Compared with normal mouse brain, mean NADH lifetimes were longer for all GBM tissues. Invasive xenografts had higher relative amounts of the longer-lived NADH component, and this correlated with decreased survival.
FLIM offers cellular resolution quantification of metabolic flux in GBM phenotypes, potentially informing biomedical researchers on improved therapeutic approaches.
多形性胶质母细胞瘤（GBM）是最常见的成人原发性脑恶性肿瘤，患者预后不良。尽管采用了积极的治疗方法，但由于治疗耐药的胶质母细胞瘤干细胞（GSCs）可能表现出代谢可塑性，GBM 仍会复发。
荧光寿命成像显微镜（FLIM）可以获得固有烟酰胺腺嘌呤二核苷酸（NADH）荧光，以检查 GSC 和 GBM 组织中其结合和游离代谢状态。
我们比较了活的人 GSC 和正常神经干细胞中 NADH 荧光寿命的平均值，并通过测量耗氧量（OCR）验证了这些结果。我们还通过测量 NADH 寿命的平均值和 NADH 长寿命成分的相对量，以及将这些结果与在原位小鼠异种移植模型中的存活情况相关联，研究了侵袭性与侵袭性较弱的 GSCs 对肿瘤代谢的作用。
GSCs 中的 NADH 荧光寿命平均值、结合态 NADH 量和 OCR 增加。与正常小鼠大脑相比，所有 GBM 组织的平均 NADH 寿命都更长。侵袭性异种移植物具有更高的长寿命 NADH 成分相对量，这与存活率降低相关。
FLIM 提供了 GBM 表型中代谢通量的细胞分辨率定量，可能为生物医学研究人员提供有关改进治疗方法的信息。
