Microfabrication Tools for In Vitro Monitoring of Cancer Cell Metabolism

描述（由申请人提供）：细胞和器官的生理或病理生理状态反映在它们的能量代谢中。例如，在肿瘤中几乎总是观察到葡萄糖代谢的偏差。癌细胞的能量代谢与正常组织明显不同，即使在有氧条件下，癌细胞的糖酵解和随后的乳酸生成也更为突出。越来越多的癌症治疗策略被设计为针对代谢偏差，使能量代谢物（如葡萄糖或乳酸）的水平与药物疗效潜在相关。本提案的目标是开发一个芯片实验室平台，用于体外监测候选药物对癌细胞能量代谢的影响。所提出的平台将紧密整合小群胶质瘤（脑癌）细胞与微型葡萄糖生物传感器在微制造设备，其中细胞微环境可以精确定义和容易调节。该平台将用于：1)在胶质瘤细胞培养中建立模拟肿瘤的微环境条件（如缺氧、酸性、营养限制）；2)用激酶信号转导通路的药物抑制剂刺激细胞；3)监测局部细胞外葡萄糖水平。因此，细胞培养/生物传感器平台将连接肿瘤微环境、能量代谢和抗癌药物疗效，并有助于描述使癌细胞更容易接受治疗的条件。该技术将为开发治疗性抗癌药物提供有价值的工具，并有助于阐明癌细胞代谢适应的分子方面。

DESCRIPTION (provided by applicant): The physiological or pathophysiological state of cells and organs is reflected in their energy metabolism. For example, deviations in glucose metabolism is almost always observed in tumors. The energy metabolism of cancer cells differs strikingly from normal tissue with glycolysis and subsequent lactate production being much more prominent in cancer cells, even under aerobic conditions. Increasingly, therapeutic strategies for cancer treatment are designed to target metabolic deviations, making levels of energy metabolites such glucose or lactate potential correlates of drug efficacy. The goal of this proposal is to develop a lab-on-chip platform for in vitro monitoring of the effects of drug candidates on energy metabolism of cancer cells. The proposed platform will intimately integrate small groups of glioma (brain cancer) cells with miniature glucose biosensors in a microfabricated device, where the cellular microenvironment can be precisely defined and easily modulated. This platform will be used to: 1) establish tumor-mimicking microenvironment conditions (e.g. hypoxic, acidic, nutrient-limiting) in glioma cell cultures; 2) challenge the cells with pharmacological inhibitors of kinase signal transduction pathways central in energy metabolism; and 3) monitor local extracellular glucose levels. Thus the cell culture/biosensor platform will connect tumor microenvironment, energy metabolism and anti-cancer drug efficacy, and will help to delineate conditions making cancer cells more susceptible to therapy. The proposed technology will be a valuable tool for the development of therapeutic anti-cancer agents, and will help illuminate molecular aspects of metabolic adaptation of cancer cells.