Autofluorescence imaging captures heterogeneous drug response differences between 2D and 3D breast cancer cultures
Autofluorescence imaging captures heterogeneous drug response differences between 2D and 3D breast cancer cultures
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DOI：
10.1364/boe.8.001911
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发表时间：
2017-03-01
影响因子：
3.4
通讯作者：
Skala, M. C.
中科院分区：
文献类型：
--
作者：
Cannon, T. M.;Shah, A. T.;Skala, M. C.
细胞自发荧光强度和寿命的双光子显微镜（光学代谢成像，或OMI）是临床前药物开发的一种有前途的工具。OMI利用来自代谢辅酶NAD（P）H和FAD的内源性荧光，对药物治疗产生的细胞代谢变化敏感。之前的研究表明，3D培养中的药物反应、基因表达、细胞间通讯和细胞信号传导与原始体内肿瘤的反应、基因表达、细胞间通讯和细胞信号传导相匹配，但与2D培养的反应、基因表达、细胞间通讯和细胞信号传导不匹配。本研究的目的是使用OMI来量化2D细胞系与由相同细胞来源的异种移植肿瘤产生的3D类器官中药物反应的动态细胞水平代谢差异。测试BT474细胞和赫赛汀抗性BT474（HR6）细胞。用载体对照、赫赛汀、XL147（PI3K抑制剂）和组合处理细胞。OMI指数用于量化响应，并且是氧化还原比（NAD（P）H的强度除以FAD）、平均NADH寿命和平均FAD寿命的线性组合。结果证实，OMI指数解决了2D与3D培养物的药物反应的显著差异（p< 0.05），特别是对于赫赛汀处理后24小时的BT474细胞，对于联合处理后24和72小时的HR6细胞，以及对于XL147处理后72小时的HR6细胞。OMI指数的细胞水平分析还揭示了对照组和处理组中处理后24、48和72小时时2D与3D培养物中细胞亚群数量的差异。最后，在对照组和所有处理组中，在处理后72小时，在2D相对于3D中测量到两种细胞系的NADH和FAD的平均寿命的显著增加（p< 0.05）。平均NADH和FAD寿命的这些全群体差异得到了2D与3D中细胞亚群数量差异的支持。总体而言，这些研究证实OMI对2D与3D中药物反应的差异敏感，并提供了有关导致这种差异的代谢细胞亚群相对丰度动态变化的进一步信息。(C)2017美国光学学会
Two-photon microscopy of cellular autofluorescence intensity and lifetime (optical metabolic imaging, or OMI) is a promising tool for preclinical drug development. OMI, which exploits the endogenous fluorescence from the metabolic coenzymes NAD(P) H and FAD, is sensitive to changes in cell metabolism produced by drug treatment. Previous studies have shown that drug response, genetic expression, cell-cell communication, and cell signaling in 3D culture match those of the original in vivo tumor, but not those of 2D culture. The goal of this study is to use OMI to quantify dynamic cell-level metabolic differences in drug response in 2D cell lines vs. 3D organoids generated from xenograft tumors of the same cell origin. BT474 cells and Herceptin-resistant BT474 (HR6) cells were tested. Cells were treated with vehicle control, Herceptin, XL147 (PI3K inhibitor), and the combination. The OMI index was used to quantify response, and is a linear combination of the redox ratio (intensity of NAD(P) H divided by FAD), mean NADH lifetime, and mean FAD lifetime. The results confirm that the OMI index resolves significant differences (p< 0.05) in drug response for 2D vs. 3D cultures, specifically for BT474 cells 24 hours after Herceptin treatment, for HR6 cells 24 and 72 hours after combination treatment, and for HR6 cells 72 hours after XL147 treatment. Cell-level analysis of the OMI index also reveals differences in the number of cell sub-populations in 2D vs. 3D culture at 24, 48, and 72 hours post-treatment in control and treated groups. Finally, significant increases (p< 0.05) in the mean lifetime of NADH and FAD were measured in 2D vs. 3D for both cell lines at 72 hours post-treatment in control and all treatment groups. These whole-population differences in the mean NADH and FAD lifetimes are supported by differences in the number of cell sub-populations in 2D vs. 3D. Overall, these studies confirm that OMI is sensitive to differences in drug response in 2D vs. 3D, and provides further information on dynamic changes in the relative abundance of metabolic cell sub-populations that contribute to this difference. (C) 2017 Optical Society of America