Elucidating the mechanism behind oscillation between glycolysis and gluconeogenesis

项目摘要 新陈代谢是一个动态的生化反应网络，能够适应变化的环境。在……里面 改变葡萄糖的可获得性、糖酵解和糖异生作用有助于维持全身的葡萄糖平衡。AS 糖酵解和糖异生操作几个常见的反应步骤，但方向相反， 热力学在糖酵解和糖异生之间的快速振荡中起着重要作用。在我们的 家长格兰特，我们做了一个有趣的观察，即使细胞系具有结构性激活的RAS和 AKT在低糖高乳酸条件下生长不佳，在振荡条件下生长迅速 在葡萄糖充足和没有葡萄糖之间的条件。我们假设糖异生的缺陷 RAS和Akt引起的酶表达使这些细胞在变化的条件和 具有讽刺意味的是，这种僵化导致他们一获得葡萄糖就迅速利用。一个 这一理论的推论是，RAS或Akt激活的细胞可能比健康细胞具有更多的同质性代谢 细胞在振荡环境中和代谢的异质性可能会破坏适应性竞争力 在不稳定波动的营养条件下。此行政补充应用程序的目标是获取 BioSpa-Cytation(Agilent Technologies)研究细胞群体内代谢异质性的作用 在振荡的营养环境中。BioSpa-Cytation是一个集孵化器和多细胞培养为一体的自动化系统 在可编程的动态环境中研究2D和3D培养的模式显微镜。最重要的是 母基金的目标是阐明振荡和协调背后的调节机制 糖酵解和糖异生通过整合代谢组学、通量组学和热力学(主要使用 质谱学、同位素示踪和定量建模)。拟议的设备将提供更多- 所需的动态细胞培养和高通量显微镜能力，使我们能够获得相干显微镜 和质谱学洞察力，并最终导致该项目在母公司的拨款中取得更大的成功。

Project Summary Metabolism is a dynamic network of biochemical reactions capable of adapting to changing environments. In changing glucose availability, glycolysis and gluconeogenesis support systemic glucose homeostasis. As glycolysis and gluconeogenesis operate several common reaction steps but in opposite directions, thermodynamics plays an important role in rapid oscillating between glycolysis and gluconeogenesis. In our parent grant, we made a fascinating observation that even though cell lines with constitutively activated Ras and Akt did not grow well under low-glucose high-lactate conditions, they proliferated rapidly under oscillatory conditions between abundant glucose and no glucose. We hypothesize that the deficiency in gluconeogenic enzyme expression caused by Ras and Akt renders these cells “inflexible” during the changing conditions and this inflexibility ironically contributes to their rapid utilization of glucose as soon as it becomes available. A corollary to this theory is that Ras- or Akt-activated cells may have more homogenous metabolism than healthy cells during oscillatory environments and the metabolic heterogeneity may undermine adaptive competitiveness in unstable fluctuating nutrient conditions. The goal of this administrative supplement application is to acquire BioSpa-Cytation (Agilent Technologies) to investigate the role of metabolic heterogeneity within cell populations in oscillatory nutrient environments. BioSpa-Cytation is an automated system that integrates incubator and multi- mode microscopy for studying 2D and 3D cultures in programmable dynamic environments. The overarching goal of the parent grant is to elucidate the regulatory mechanisms behind oscillation and coordination between glycolysis and gluconeogenesis by integrating metabolomics, fluxomics, and thermodynamics (mainly using mass spectrometry, isotope tracing, and quantitative modeling). The proposed equipment will provide the much- needed dynamic cell culturing and high-throughput microscopy capabilities, allow us to gain coherent microscopy and mass spectrometry insights, and ultimately lead to a greater success of the project in the parent grant.