Cell-based assays using bioelectronic sensor-chips for the dynamic analysis of tumor cell metabolism and chemosensitivity

了解肿瘤细胞在不同微环境中的能量代谢调控不仅对肿瘤的诊断和治疗，而且对更有效地应用（化学）治疗策略至关重要。为了实现这一目标，本项目的目的是系统地分析肿瘤细胞培养物中的基本代谢参数，这些参数是在代表不同肿瘤微环境的特定条件下选择的，例如高酸度和缺氧。实验装置由多参数微传感器芯片组成，其连续和并行地监测细胞培养物的pH、pO2（代表代谢活动）和阻抗（反映膜相关特性）的变化。流体灌注系统不仅模拟选定的微生理条件，而且维持长期孵育的条件。两种乳腺癌细胞系将作为不同恶性肿瘤的模型。对于肿瘤细胞代谢的化学治疗调节，将在微环境参数的不同组合下测试指定用于临床使用的糖酵解和谷氨酸代谢抑制剂的功效。这些结果不仅为未来肿瘤代谢的系统生物学分析提供了数据，而且为设计更好的诊断和更有效的肿瘤治疗方法提供了新的理论基础。此外，与微传感器芯片获得的经验，应有助于完善和实现生物电子芯片系统，进一步应用于细胞的功能分析。

Understandig the modulations in energy metabolism of tumor cells in a variable microenvironment is essential not only for the diagnosis and prognoses of cancers, but also for a more efficient application of (chemo)therapeutic strategies. Towards this goal, it is the aim of this project to systematically analyze basic metabolic parameters in tumor cell cultures under a selection of defined conditions representing different tumor microenvironment, such as high acidity, and hypoxia. The experimental set-up is composed of multiparametric microsensorchips which monitor changes in pH, pO2 (representing metabolic activities) and impedance (reflecting membrane-associated properties) of cell cultures continuously and in parallel. A fluidic perfusion system not only simulates selected microphysiological conditions, but also maintains conditions for long term incubations. Two breast cancer cell lines will serve as a model for tumors of different malignancy. For chemotherapeutic modulations of tumor cell metabolisms, inhibitors of glycolysis and glutamate metabolism designated for clinical use are to be tested for their efficacy under different combinations of microenvironmental parameters. The results provide data not only for future systems biological analysis of tumor metabolism but also for developing new rationales in designing better diagnostics and more efficient therapeutic approaches in oncology. Moreover, the experience obtained with the microsensor chips should serve to refine and implement bioelectronic chip system for further applications in the functional analysis of cells.