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评估谷氨酸吸收的超分子探针使单细胞中的半量化代谢模型成为可能
Min Xue1, Wei Wei2, Yapeng Su1
1Division of Chemistry and Chemical Engineering, California Institute of Technology , Pasadena, California 91125, United States.
Journal of the American Chemical Society
|February 27, 2016
概括
我们开发了一种测量单细胞胺摄取量的新方法. 这种方法揭示了癌细胞如何适应其能量代谢和信号通路,以应对EGFR抑制剂等向治疗.
科学领域:
- 生物化学
- 细胞生物学
- 癌症研究
背景情况:
- 在单个细胞中量化营养吸收对于了解细胞代谢和疾病至关重要.
- 现有的方法往往缺乏分析复杂细胞过程的灵敏度或多重复合能力.
- 受体氨酸激酶 (RTK) 信号传递在癌症中起着关键作用,但其与细胞能量代谢的相互作用仍然不完全理解.
研究的目的:
- 开发和验证一种新的超分子表面竞争试验,以精确量化单细胞中的谷氨酸摄取量.
- 将此测试与单细胞条码芯片上的其他测量进行整合,以建模细胞能量潜力.
- 研究EGFR抑制对脑癌细胞代谢和信号的作用.
主要方法:
- 用于基于FRET的检测,采用了使用Cy3标记的环氧和阿达曼坦-BHQ2合物的超分子宿主/客体系统.
- 一种标有阿达曼坦的谷氨酸类比物已被确定并用于细胞吸收研究.
- 该测试被集成到一个单细胞条形码芯片中,测量了15种其他代谢物,酶和蛋白,并输入到稳定状态模型中.
主要成果:
- 该试验成功量化了单细胞中的谷氨酸摄取量.
- 与其他测量相结合,可以建模细胞能量潜力及其与RTK信号的相关性.
- 在模型脑癌细胞中,低剂量欧洛替尼治疗 (1μM) 矛盾地增加了细胞能量潜力,尽管抑制了葡萄糖吸收和蛋白信号传递.
结论:
- 开发的试验为剖析单细胞代谢动态提供了强大的工具.
- 在EGFR抑制后观察到意想不到的代谢适应,突出显示了复杂的调节机制.
- 发现了蛋白信号传递和细胞能量代谢之间的新相互作用,这可能解释了癌症患者对EGFR抑制剂的耐药性.
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