Elucidating the mechanism behind oscillation between glycolysis and gluconeogenesis

阐明糖酵解和糖异生之间振荡背后的机制

• 批准号: 10799353
• 负责人: Junyoung O. Park
• 金额: $ 24.42万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10799353
• 关键词: Administrative Supplement; Biochemical Reaction; Cell Line; Cells; Environment; Enzymes; Equipment; Gluconeogenesis; Glucose; Glycolysis; Goals; Heterogeneity; Human; Incubators; Isotopes; Mass Spectrum Analysis; Metabolic; Metabolic Pathway; Metabolism; Microscopy; Modeling; Molecular; Nutrient; Play; Population; Proliferating; Reaction; Role; System; Technology; Testing; Thermodynamics; Visual; blood glucose regulation; cytate; fascinate; insight; metabolomics; parent grant; success; theories; three dimensional cell culture

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.

项目摘要 新陈代谢是一个动态的生化反应网络，能够适应不断变化的环境。在 改变葡萄糖的利用率、糖酵解和糖异生支持全身葡萄糖稳态。作为 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重要作用。在我们 在父母的资助下，我们做了一个有趣的观察，即使细胞系具有组成性激活的Ras和 Akt在低葡萄糖高乳酸条件下生长不好，它们在振荡条件下迅速增殖 在大量葡萄糖和无葡萄糖之间的条件。我们假设， 由Ras和Akt引起的酶表达使这些细胞在变化的条件下“不灵活”， 具有讽刺意味的是，一旦葡萄糖变得可用，这种可溶解性有助于它们快速利用葡萄糖。一 这一理论的一个推论是Ras或Akt激活的细胞可能比健康的细胞具有更均匀的代谢， 细胞在振荡环境和代谢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。这一行政补充申请的目的是获得 BioSpa-Cytation（Agilent Technologies）研究细胞群内代谢异质性的作用 在振荡的营养环境中。BioSpa-Cytation是一种自动化系统，集成了孵育器和多个 模式显微镜用于在可编程动态环境中研究2D和3D文化。总体 父母补助金的目标是阐明振荡和协调背后的调节机制， 通过整合代谢组学，通量组学和热力学（主要使用 质谱、同位素示踪和定量建模）。拟议的设备将提供更多- 需要动态细胞培养和高通量显微镜功能，使我们能够获得连贯的显微镜检查 和质谱分析的见解，并最终导致该项目在母基金中取得更大的成功。