Characterization of an Aggregated Three-Dimensional Cell Culture Model by Multimodal Mass Spectrometry Imaging
Characterization of an Aggregated Three-Dimensional Cell Culture Model by Multimodal Mass Spectrometry Imaging
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DOI：
10.1021/acs.analchem.0c02389
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发表时间：
2020-09-15
影响因子：
7.4
通讯作者：
Clench, Malcolm R.
中科院分区：
文献类型：
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作者：
Flint, Lucy E.;Hamm, Gregory;Clench, Malcolm R.
质谱成像（MSI）是一种成熟的分析工具，能够通过确定生物分子的空间分布来定义和理解复杂组织。三维（3D）细胞培养模型模拟体内肿瘤的病理生理环境，并迅速成为一种有价值的研究工具。在这里，多模式MSI技术被用来表征一种新的聚集的3D肺腺癌模型，该模型由该小组开发，以模拟体内组织。基于多种内源性分子的空间分布，观察肿瘤异质性区域和缺氧微环境。解吸电喷雾电离（DESI）-MSI从代谢物分布中定义了缺氧核心和增殖外层的区域。靶向代谢物（例如，乳酸盐、谷氨酰胺和柠檬酸盐）映射到糖酵解和TCA循环的途径，证明肿瘤代谢行为。首次应用成像质量细胞仪（IMC）与三维细胞培养，使单细胞表型在1 μ m的空间分辨率。增殖（Ki-67）和缺氧（葡萄糖转运蛋白1）的蛋白质标志物定义了类聚集体模型中的代谢信号，补充了代谢物数据。激光消融电感耦合等离子体（LA-ICP）-MSI分析定位内源性元素，包括镁和铜，进一步区分缺氧梯度和验证蛋白质表达。获得大量关于互补性质的分子信息使得能够深入了解新型肿瘤模型中的生物学过程。结合强大的成像技术来表征聚集的3D培养物，突出了在癌症研究和药物开发中具有潜在应用的未来方法。
Mass spectrometry imaging (MSI) is an established analytical tool capable of defining and understanding complex tissues by determining the spatial distribution of biological molecules. Three-dimensional (3D) cell culture models mimic the pathophysiological environment of in vivo tumors and are rapidly emerging as a valuable research tool. Here, multimodal MSI techniques were employed to characterize a novel aggregated 3D lung adenocarcinoma model, developed by the group to mimic the in vivo tissue. Regions of tumor heterogeneity and the hypoxic microenvironment were observed based on the spatial distribution of a variety of endogenous molecules. Desorption electrospray ionization (DESI)-MSI defined regions of a hypoxic core and a proliferative outer layer from metabolite distribution. Targeted metabolites (e.g., lactate, glutamine, and citrate) were mapped to pathways of glycolysis and the TCA cycle demonstrating tumor metabolic behavior. The first application of imaging mass cytometry (IMC) with 3D cell culture enabled single-cell phenotyping at 1 mu m spatial resolution. Protein markers of proliferation (Ki-67) and hypoxia (glucose transporter 1) defined metabolic signaling in the aggregoid model, which complemented the metabolite data. Laser ablation inductively coupled plasma (LA-ICP)-MSI analysis localized endogenous elements including magnesium and copper, further differentiating the hypoxia gradient and validating the protein expression. Obtaining a large amount of molecular information on a complementary nature enabled an in-depth understanding of the biological processes within the novel tumor model. Combining powerful imaging techniques to characterize the aggregated 3D culture highlighted a future methodology with potential applications in cancer research and drug development.