Metabolomics in postmortem cerebrospinal fluid diagnostics: a state-of-the-art method to interpret central nervous system–related pathological processes

Bohnert, Simone; Reinert, Christoph; Trella, Stefanie; Schmitz, W.; Ondruschka, Benjamin; Bohnert, Michael

doi:10.1007/s00414-020-02462-2

Cited by 20 publications

(14 citation statements)

References 45 publications

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“…As already published by us, MetR efficiently results in a characteristic metabolic fingerprint and footprint in L929 cells [ 22 ]. The footprint represents a small group of metabolites, which allows a specific definition of AR.…”

Section: Resultsmentioning

confidence: 84%

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Cysteine Restriction in Murine L929 Fibroblasts as an Alternative Strategy to Methionine Restriction in Cancer Therapy

Schmitz¹,

Ries

Koderer

et al. 2021

IJMS

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Methionine restriction (MetR) is an efficient method of amino acid restriction (AR) in cells and organisms that induces low energy metabolism (LEM) similar to caloric restriction (CR). The implementation of MetR as a therapy for cancer or other diseases is not simple since the elimination of a single amino acid in the diet is difficult. However, the in vivo turnover rate of cysteine is usually higher than the rate of intake through food. For this reason, every cell can enzymatically synthesize cysteine from methionine, which enables the use of specific enzymatic inhibitors. In this work, we analysed the potential of cysteine restriction (CysR) in the murine cell line L929. This study determined metabolic fingerprints using mass spectrometry (LC/MS). The profiles were compared with profiles created in an earlier work under MetR. The study was supplemented by proliferation studies using D-amino acid analogues and inhibitors of intracellular cysteine synthesis. CysR showed a proliferation inhibition potential comparable to that of MetR. However, the metabolic footprints differed significantly and showed that CysR does not induce classic LEM at the metabolic level. Nevertheless, CysR offers great potential as an alternative for decisive interventions in general and tumour metabolism at the metabolic level.

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Section: Resultsmentioning

confidence: 84%

“…L929 cells react rapidly and efficiently to MetR, and an analysis of more than 150 different metabolites belonging to different classes (amino acids, urea and TCA cycles, carbohydrates, etc.) by LC/MS defined a metabolic fingerprint and enabled the identification of specific metabolites reflecting normal or MetR (LEM) conditions [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

Cysteine Restriction in Murine L929 Fibroblasts as an Alternative Strategy to Methionine Restriction in Cancer Therapy

Schmitz¹,

Ries

Koderer

et al. 2021

IJMS

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“…In principle, CSF, which communicates without barriers with the extracellular space of the central nervous system (CNS), i.e., with the milieu surrounding the neurons and glial cells, seems to be suitable to reflect central nervous processes such as biochemical processes in the brain after traumatization (“neuroforensomics”), not only because of its apparent postmortem stability [ 43 ]. In addition, this makes it possible to investigate CSF even in cases without visible signs of impact, and it appears promising in the light of the results presented here to identify a possible central nervous involvement and to help clarify controversial causes and circumstances of death.…”

Section: Discussionmentioning

confidence: 99%

Myelin basic protein and neurofilament H in postmortem cerebrospinal fluid as surrogate markers of fatal traumatic brain injury

et al. 2021

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The aim of this study was to investigate if the biomarkers myelin basic protein (MBP) and neurofilament-H (NF-H) yielded informative value in forensic diagnostics when examining cadaveric cerebrospinal fluid (CSF) biochemically via an enzyme-linked immunosorbent assay (ELISA) and comparing the corresponding brain tissue in fatal traumatic brain injury (TBI) autopsy cases by immunocytochemistry versus immunohistochemistry. In 21 trauma and 19 control cases, CSF was collected semi-sterile after suboccipital puncture and brain specimens after preparation. The CSF MBP (p = 0.006) and NF-H (p = 0.0002) levels after TBI were significantly higher than those in cardiovascular controls. Immunohistochemical staining against MBP and against NF-H was performed on cortical and subcortical samples from also biochemically investigated cases (5 TBI cases/5 controls). Compared to the controls, the TBI cases showed a visually reduced staining reaction against MBP or repeatedly ruptured neurofilaments against NF-H. Immunocytochemical tests showed MBP-positive phagocytizing macrophages in CSF with a survival time of > 24 h. In addition, numerous TMEM119-positive microglia could be detected with different degrees of staining intensity in the CSF of trauma cases. As a result, we were able to document that elevated levels of MBP and NF-H in the CSF should be considered as useful neuroinjury biomarkers of traumatic brain injury.

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“…LC/MS analyses were carried out as published before [ 39 , 40 ] in four independent experiments at 24 h, 48 h, 72 h, 96 h and 120 h, each value in triplicate. Metabolites were quantified in cell pellets and corresponding supernatants (media) under methionine-containing and methionine-free conditions (12 samples per time point in total).…”

Section: Methodsmentioning

confidence: 99%

Metabolic Fingerprinting of Murine L929 Fibroblasts as a Cell-Based Tumour Suppressor Model System for Methionine Restriction

Schmitz¹,

Koderer

El-Mesery

et al. 2021

IJMS

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Since Otto Warburg reported in 1924 that cancer cells address their increased energy requirement through a massive intake of glucose, the cellular energy level has offered a therapeutic anticancer strategy. Methionine restriction (MetR) is one of the most effective approaches for inducing low-energy metabolism (LEM) due to the central position in metabolism of this amino acid. However, no simple in vitro system for the rapid analysis of MetR is currently available, and this study establishes the murine cell line L929 as such a model system. L929 cells react rapidly and efficiently to MetR, and the analysis of more than 150 different metabolites belonging to different classes (amino acids, urea and tricarboxylic acid cycle (TCA) cycles, carbohydrates, etc.) by liquid chromatography/mass spectrometry (LC/MS) defines a metabolic fingerprint and enables the identification of specific metabolites representing normal or MetR conditions. The system facilitates the rapid and efficient testing of potential cancer therapeutic metabolic targets. To date, MS studies of MetR have been performed using organisms and yeast, and the current LC/MS analysis of the intra- and extracellular metabolites in the murine cell line L929 over a period of 5 days thus provides new insights into the effects of MetR at the cellular metabolic level.

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Metabolomics in postmortem cerebrospinal fluid diagnostics: a state-of-the-art method to interpret central nervous system–related pathological processes

Cited by 20 publications

References 45 publications

Cysteine Restriction in Murine L929 Fibroblasts as an Alternative Strategy to Methionine Restriction in Cancer Therapy

Cysteine Restriction in Murine L929 Fibroblasts as an Alternative Strategy to Methionine Restriction in Cancer Therapy

Myelin basic protein and neurofilament H in postmortem cerebrospinal fluid as surrogate markers of fatal traumatic brain injury

Metabolic Fingerprinting of Murine L929 Fibroblasts as a Cell-Based Tumour Suppressor Model System for Methionine Restriction

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