13C-metabolic flux analysis reveals metabolic rewiring in HL-60 neutrophil-like cells through differentiation and immune stimulation

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Takeo Taniguchi , Nobuyuki Okahashi , Fumio Matsuda
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引用次数: 0

Abstract

Neutrophils are innate immune cells and the first line of defense for the maintenance of homeostasis. However, our knowledge of the metabolic rewiring associated with their differentiation and immune stimulation is limited. Here, quantitative 13C-metabolic flux analysis was performed using HL-60 cells as the neutrophil model. A metabolic model for 13C-metabolic flux analysis of neutrophils was developed based on the accumulation of 13C in intracellular metabolites derived from 13C-labeled extracellular carbon sources and intracellular macromolecules. Aspartate and glutamate in the medium were identified as carbon sources that enter central carbon metabolism. Furthermore, the breakdown of macromolecules, estimated to be fatty acids and nucleic acids, was observed. Based on these results, a modified metabolic model was used for 13C-metabolic flux analysis of undifferentiated, differentiated, and lipopolysaccharide (LPS)-activated HL-60 cells. The glucose uptake rate and glycolytic flux decreased with differentiation, whereas the tricarboxylic acid (TCA) cycle flux remained constant. The addition of LPS to differentiated HL-60 cells activated the glucose uptake rate and pentose phosphate pathway (PPP) flux levels, resulting in an increased rate of total NADPH regeneration, which could be used to generate reactive oxygen species. The flux levels of fatty acid degradation and synthesis were also increased in LPS-activated HL-60 cells. Overall, this study highlights the quantitative metabolic alterations in multiple pathways via the differentiation and activation of HL-60 cells using 13C-metabolic flux analysis.

13C 代谢通量分析揭示了 HL-60 中性粒细胞样细胞在分化和免疫刺激过程中的代谢重构
中性粒细胞是先天性免疫细胞,也是维持体内平衡的第一道防线。然而,我们对与中性粒细胞分化和免疫刺激相关的新陈代谢重新布线了解有限。在此,我们以 HL-60 细胞为中性粒细胞模型进行了 13C 代谢通量定量分析。中性粒细胞 13C 代谢通量分析的代谢模型是基于 13C 标记的细胞外碳源和细胞内大分子在细胞内代谢产物中的积累而建立的。培养基中的天冬氨酸和谷氨酸被确定为进入中枢碳代谢的碳源。此外,还观察到大分子的分解,估计是脂肪酸和核酸。在这些结果的基础上,对未分化、已分化和脂多糖(LPS)活化的 HL-60 细胞进行了 13C 代谢通量分析。葡萄糖摄取率和糖酵解通量随细胞分化而降低,而三羧酸(TCA)循环通量则保持不变。向分化的 HL-60 细胞中添加 LPS 会激活葡萄糖摄取率和磷酸戊糖途径通量水平,导致 NADPH 总再生率增加,从而产生活性氧。在 LPS 激活的 HL-60 细胞中,脂肪酸降解和合成的通量水平也有所增加。总之,本研究通过 13C 代谢通量分析,强调了 HL-60 细胞分化和活化过程中多种途径的定量代谢变化。
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来源期刊
Metabolic Engineering Communications
Metabolic Engineering Communications Medicine-Endocrinology, Diabetes and Metabolism
CiteScore
13.30
自引率
1.90%
发文量
22
审稿时长
18 weeks
期刊介绍: Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.
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