Metabolic profiling of single cells by exploiting NADH and FAD fluorescence via flow cytometry

IF 7 2区医学 Q1 ENDOCRINOLOGY & METABOLISM Molecular Metabolism Pub Date : 2024-07-04 DOI:10.1016/j.molmet.2024.101981

Ariful Haque Abir , Leonie Weckwerth , Artur Wilhelm , Jana Thomas , Clara M. Reichardt , Luis Munoz , Simon Völkl , Uwe Appelt , Markus Mroz , Raluca Niesner , Anja Hauser , Rebecca Sophie Fischer , Katharina Pracht , Hans-Martin Jäck , Georg Schett , Gerhard Krönke , Dirk Mielenz

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引用次数: 0

Abstract

Objective

The metabolism of different cells within the same microenvironment can differ and dictate physiological or pathological adaptions. Current single-cell analysis methods of metabolism are not label-free.

Methods

The study introduces a label-free, live-cell analysis method assessing endogenous fluorescence of NAD(P)H and FAD in surface-stained cells by flow cytometry.

Results

OxPhos inhibition, mitochondrial uncoupling, glucose exposure, genetic inactivation of glucose uptake and mitochondrial respiration alter the optical redox ratios of FAD and NAD(P)H as measured by flow cytometry. Those alterations correlate strongly with measurements obtained by extracellular flux analysis. Consequently, metabolically distinct live B-cell populations can be resolved, showing that human memory B-cells from peripheral blood exhibit a higher glycolytic flexibility than naïve B cells. Moreover, the comparison of blood-derived B- and T-lymphocytes from healthy donors and rheumatoid arthritis patients unleashes rheumatoid arthritis-associated metabolic traits in human naïve and memory B-lymphocytes.

Conclusions

Taken together, these data show that the optical redox ratio can depict metabolic differences in distinct cell populations by flow cytometry.

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通过流式细胞仪利用 NADH 和 FAD 荧光绘制单细胞代谢图谱。

同一微环境中不同细胞的新陈代谢可能不同，并决定生理或病理适应。目前的单细胞新陈代谢分析方法并非无标记。本研究介绍了一种无标记的活细胞分析方法，通过流式细胞仪评估表面染色细胞中 NAD(P)H 和 FAD 的内源性荧光。通过流式细胞仪测量，OxPhos 抑制、线粒体解偶联、葡萄糖暴露、葡萄糖摄取和线粒体呼吸的遗传失活会改变 FAD 和 NAD(P)H 的光学氧化还原比率。这些变化与细胞外通量分析的测量结果密切相关。结果表明，外周血中的人类记忆 B 细胞比天真 B 细胞表现出更高的糖酵解灵活性。此外，通过比较健康捐献者和类风湿性关节炎患者血液中的 B 淋巴细胞和 T 淋巴细胞，还发现了人类幼稚 B 淋巴细胞和记忆 B 淋巴细胞与类风湿性关节炎相关的代谢特征。总之，这些数据表明，光学氧化还原比率可以通过流式细胞术描述不同细胞群的代谢差异。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Metabolism ENDOCRINOLOGY & METABOLISM-

CiteScore

14.50

自引率

2.50%

发文量

219

审稿时长

43 days

期刊介绍： Molecular Metabolism is a leading journal dedicated to sharing groundbreaking discoveries in the field of energy homeostasis and the underlying factors of metabolic disorders. These disorders include obesity, diabetes, cardiovascular disease, and cancer. Our journal focuses on publishing research driven by hypotheses and conducted to the highest standards, aiming to provide a mechanistic understanding of energy homeostasis-related behavior, physiology, and dysfunction. We promote interdisciplinary science, covering a broad range of approaches from molecules to humans throughout the lifespan. Our goal is to contribute to transformative research in metabolism, which has the potential to revolutionize the field. By enabling progress in the prognosis, prevention, and ultimately the cure of metabolic disorders and their long-term complications, our journal seeks to better the future of health and well-being.