Summary of Study ST004020

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002515. The data can be accessed directly via it's Project DOI: 10.21228/M8JN9T This work is supported by NIH grant, U2C- DK119886.

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This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST004020
Study TitleAnalysis of the effects of MINCH on the metabolism of human preadipocytes and mature adipocytes by 13C metabolic tracing with [1,2-13C]glucose
Study SummaryIn the second part of the project, the observed effects of MINCH on glycolysis in human preadipocytes and mature adipocytes were validated with [1,2-13C]glucose. Again, the effects were compared with rosiglitazone-treated cells and untreated control cells. The [1,2-13C]-glucose labeling data confirmed the increased glycolytic and PPP activity in MINCH-treated preadipocytes and mature adipocytes. Analysis of the PPP contribution to overall glucose metabolism (i.e., glycolysis and PPP) showed no increase in mature adipocytes and even a decrease in preadipocytes, in contrast to rosiglitazone treatment, indicating an increased glycolytic contribution to glucose metabolism by MINCH treatment. This suggests that MINCH produces its effects in part via a PPARG-independent mechanism, which is consistent with previous reports.
Institute
Helmholtz Centre for Environmental Research
Last NameGoerdeler
First NameCornelius
AddressPermoserstr. 15
Emailcornelius.goerdeler@ufz.de
Phone004934160252713
Submit Date2025-06-18
Raw Data AvailableYes
Raw Data File Type(s)mzML, wiff
Analysis Type DetailLC-MS
Release Date2025-07-24
Release Version1
Cornelius Goerdeler Cornelius Goerdeler
https://dx.doi.org/10.21228/M8JN9T
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002515
Project DOI:doi: 10.21228/M8JN9T
Project Title:13C Metabolic Tracing in Human SGBS Cells Provides a Potential New Approach Methodology for Assessing Metabolism-Disrupting Properties
Project Summary:Due to the increased use and production of plastic materials worldwide, humans are ubiquitously exposed to plastic additives, including plasticizers. Recent research suggests that exposure to certain plasticizers promotes obesity due to their metabolism-disrupting properties. Following the ban on di-(2-ethylhexyl) phthalate (DEHP) and other phthalate plasticizers due to their reproductive toxicity, substitutes such as the plasticizer diisononylcyclohexane-1,2-dicarboxylate (DINCH) have been increasingly used. However, in vitro, studies indicate that the primary metabolite monoisononylcyclohexane-1,2-dicarboxylic acid ester (MINCH) promotes differentiation of human adipocytes. In contrast, no obesogenic effect has been observed in application studies in vivo. The absence of weight-promoting effects of DINCH was confirmed in a recent study with DINCH-exposed C57BL/6N mice, but an increase in adipocyte size in visceral adipose tissue and sex-specific effects on serum lipid levels together with impaired insulin sensitivity were observed. Therefore, as there is still limited information about the potential metabolism-disrupting properties of MINCH, we used 13C tracing as a novel method to investigate the effects of MINCH on the pathway activity of central carbon metabolism in human adipocytes. In contrast to metabolomics, which provides information on changes in the abundance of metabolites, 13C metabolic tracing provides an overview of changes in metabolic pathway activity, enabling an in-depth understanding of how metabolism-disrupting chemicals might disrupt cellular metabolism. The changes in central carbon metabolism activity following MINCH treatment were analyzed after insulin stimulation using three carbon tracers. The project consists of three main studies, depending on the use of the carbon tracer: 1. Analysis of the effects of MINCH on glycolysis and pentose phosphate pathway (PPP) activity, the contribution of glucose to the tricarboxylic acid cycle (TCA) and pyruvate carboxylase-mediated anaplerosis in human preadipocytes and mature adipocytes using [U-13C]glucose; 2. Validation of the effects of MINCH on glycolysis and PPP activity and discrimination of their contribution to glucose metabolism in human preadipocytes and mature adipocytes using [1,2-13C]glucose; 3. Assessment of glyceroneogenesis activity, glutamine contribution to the TCA cycle, oxidative flux through the TCA, reductive carboxylation via isocitrate dehydrogenase (IDH) for lipid synthesis and cycling of metabolites through the TCA cycle in human preadipocytes and mature adipocytes using [U-13C]-glutamine.
Institute:Helmholtz Centre for Environmental Research
Last Name:Engelmann
First Name:Beatrice
Address:Permoserstr. 15
Email:beatrice.engelmann@ufz.de
Phone:004934160251099

Subject:

Subject ID:SU004158
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Gender:Male

Factors:

Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Sample source Treatment Sample Type Labeling time
SA462258Adult_Ctrl_24h_1_2Glc_1SGBS mature adipocytes Control cell pellet 24 h
SA462259Adult_Ctrl_24h_1_2Glc_3SGBS mature adipocytes Control cell pellet 24 h
SA462260Adult_Ctrl_24h_1_2Glc_4SGBS mature adipocytes Control cell pellet 24 h
SA462261Adult_Ctrl_24h_1_2Glc_2SGBS mature adipocytes Control cell pellet 24 h
SA462262Adult_Ctrl_3h_1_2Glc_4SGBS mature adipocytes Control cell pellet 3 h
SA462263Adult_Ctrl_3h_1_2Glc_3SGBS mature adipocytes Control cell pellet 3 h
SA462264Adult_Ctrl_3h_1_2Glc_2SGBS mature adipocytes Control cell pellet 3 h
SA462265Adult_Ctrl_3h_1_2Glc_1SGBS mature adipocytes Control cell pellet 3 h
SA462266Adult_Ctrl_SN_24h_1_2Glc_4SGBS mature adipocytes Control cell supernatant 24 h
SA462267Adult_Ctrl_SN_24h_1_2Glc_3SGBS mature adipocytes Control cell supernatant 24 h
SA462268Adult_Ctrl_SN_24h_1_2Glc_2SGBS mature adipocytes Control cell supernatant 24 h
SA462269Adult_Ctrl_SN_24h_1_2Glc_1SGBS mature adipocytes Control cell supernatant 24 h
SA462270Adult_Ctrl_SN_3h_1_2Glc_1SGBS mature adipocytes Control cell supernatant 3 h
SA462271Adult_Ctrl_SN_3h_1_2Glc_2SGBS mature adipocytes Control cell supernatant 3 h
SA462272Adult_Ctrl_SN_3h_1_2Glc_3SGBS mature adipocytes Control cell supernatant 3 h
SA462273Adult_Ctrl_SN_3h_1_2Glc_4SGBS mature adipocytes Control cell supernatant 3 h
SA462274Adult_DINCH_10uM_24h_1_2Glc_3SGBS mature adipocytes DINCH 10µM cell pellet 24 h
SA462275Adult_DINCH_10uM_24h_1_2Glc_2SGBS mature adipocytes DINCH 10µM cell pellet 24 h
SA462276Adult_DINCH_10uM_24h_1_2Glc_4SGBS mature adipocytes DINCH 10µM cell pellet 24 h
SA462277Adult_DINCH_10uM_24h_1_2Glc_1SGBS mature adipocytes DINCH 10µM cell pellet 24 h
SA462278Adult_DINCH_10uM_3h_1_2Glc_4SGBS mature adipocytes DINCH 10µM cell pellet 3 h
SA462279Adult_DINCH_10uM_3h_1_2Glc_3SGBS mature adipocytes DINCH 10µM cell pellet 3 h
SA462280Adult_DINCH_10uM_3h_1_2Glc_2SGBS mature adipocytes DINCH 10µM cell pellet 3 h
SA462281Adult_DINCH_10uM_3h_1_2Glc_1SGBS mature adipocytes DINCH 10µM cell pellet 3 h
SA462282Adult_DINCH_10uM_SN_24h_1_2Glc_3SGBS mature adipocytes DINCH 10µM cell supernatant 24 h
SA462283Adult_DINCH_10uM_SN_24h_1_2Glc_4SGBS mature adipocytes DINCH 10µM cell supernatant 24 h
SA462284Adult_DINCH_10uM_SN_24h_1_2Glc_2SGBS mature adipocytes DINCH 10µM cell supernatant 24 h
SA462285Adult_DINCH_10uM_SN_24h_1_2Glc_1SGBS mature adipocytes DINCH 10µM cell supernatant 24 h
SA462286Adult_DINCH_10uM_SN_3h_1_2Glc_2SGBS mature adipocytes DINCH 10µM cell supernatant 3 h
SA462287Adult_DINCH_10uM_SN_3h_1_2Glc_3SGBS mature adipocytes DINCH 10µM cell supernatant 3 h
SA462288Adult_DINCH_10uM_SN_3h_1_2Glc_4SGBS mature adipocytes DINCH 10µM cell supernatant 3 h
SA462289Adult_DINCH_10uM_SN_3h_1_2Glc_1SGBS mature adipocytes DINCH 10µM cell supernatant 3 h
SA462290Adult_MINCH_10nM_24h_1_2Glc_1SGBS mature adipocytes MINCH 10nM cell pellet 24 h
SA462291Adult_MINCH_10nM_24h_1_2Glc_2SGBS mature adipocytes MINCH 10nM cell pellet 24 h
SA462292Adult_MINCH_10nM_24h_1_2Glc_3SGBS mature adipocytes MINCH 10nM cell pellet 24 h
SA462293Adult_MINCH_10nM_24h_1_2Glc_4SGBS mature adipocytes MINCH 10nM cell pellet 24 h
SA462294Adult_MINCH_10nM_3h_1_2Glc_2SGBS mature adipocytes MINCH 10nM cell pellet 3 h
SA462295Adult_MINCH_10nM_3h_1_2Glc_1SGBS mature adipocytes MINCH 10nM cell pellet 3 h
SA462296Adult_MINCH_10nM_3h_1_2Glc_4SGBS mature adipocytes MINCH 10nM cell pellet 3 h
SA462297Adult_MINCH_10nM_3h_1_2Glc_3SGBS mature adipocytes MINCH 10nM cell pellet 3 h
SA462298Adult_MINCH_10nM_SN_24h_1_2Glc_3SGBS mature adipocytes MINCH 10nM cell supernatant 24 h
SA462299Adult_MINCH_10nM_SN_24h_1_2Glc_1SGBS mature adipocytes MINCH 10nM cell supernatant 24 h
SA462300Adult_MINCH_10nM_SN_24h_1_2Glc_2SGBS mature adipocytes MINCH 10nM cell supernatant 24 h
SA462301Adult_MINCH_10nM_SN_24h_1_2Glc_4SGBS mature adipocytes MINCH 10nM cell supernatant 24 h
SA462302Adult_MINCH_10nM_SN_3h_1_2Glc_1SGBS mature adipocytes MINCH 10nM cell supernatant 3 h
SA462303Adult_MINCH_10nM_SN_3h_1_2Glc_4SGBS mature adipocytes MINCH 10nM cell supernatant 3 h
SA462304Adult_MINCH_10nM_SN_3h_1_2Glc_3SGBS mature adipocytes MINCH 10nM cell supernatant 3 h
SA462305Adult_MINCH_10nM_SN_3h_1_2Glc_2SGBS mature adipocytes MINCH 10nM cell supernatant 3 h
SA462306Adult_MINCH_10uM_24h_1_2Glc_1SGBS mature adipocytes MINCH 10µM cell pellet 24 h
SA462307Adult_MINCH_10uM_24h_1_2Glc_2SGBS mature adipocytes MINCH 10µM cell pellet 24 h
SA462308Adult_MINCH_10uM_24h_1_2Glc_4SGBS mature adipocytes MINCH 10µM cell pellet 24 h
SA462309Adult_MINCH_10uM_24h_1_2Glc_3SGBS mature adipocytes MINCH 10µM cell pellet 24 h
SA462310Adult_MINCH_10uM_3h_1_2Glc_4SGBS mature adipocytes MINCH 10µM cell pellet 3 h
SA462311Adult_MINCH_10uM_3h_1_2Glc_2SGBS mature adipocytes MINCH 10µM cell pellet 3 h
SA462312Adult_MINCH_10uM_3h_1_2Glc_3SGBS mature adipocytes MINCH 10µM cell pellet 3 h
SA462313Adult_MINCH_10uM_3h_1_2Glc_1SGBS mature adipocytes MINCH 10µM cell pellet 3 h
SA462314Adult_MINCH_10uM_SN_24h_1_2Glc_3SGBS mature adipocytes MINCH 10µM cell supernatant 24 h
SA462315Adult_MINCH_10uM_SN_24h_1_2Glc_2SGBS mature adipocytes MINCH 10µM cell supernatant 24 h
SA462316Adult_MINCH_10uM_SN_24h_1_2Glc_1SGBS mature adipocytes MINCH 10µM cell supernatant 24 h
SA462317Adult_MINCH_10uM_SN_24h_1_2Glc_4SGBS mature adipocytes MINCH 10µM cell supernatant 24 h
SA462318Adult_MINCH_10uM_SN_3h_1_2Glc_1SGBS mature adipocytes MINCH 10µM cell supernatant 3 h
SA462319Adult_MINCH_10uM_SN_3h_1_2Glc_3SGBS mature adipocytes MINCH 10µM cell supernatant 3 h
SA462320Adult_MINCH_10uM_SN_3h_1_2Glc_2SGBS mature adipocytes MINCH 10µM cell supernatant 3 h
SA462321Adult_MINCH_10uM_SN_3h_1_2Glc_4SGBS mature adipocytes MINCH 10µM cell supernatant 3 h
SA462322Adult_Rosi_24h_1_2Glc_1SGBS mature adipocytes Rosiglitazone cell pellet 24 h
SA462323Adult_Rosi_24h_1_2Glc_2SGBS mature adipocytes Rosiglitazone cell pellet 24 h
SA462324Adult_Rosi_24h_1_2Glc_3SGBS mature adipocytes Rosiglitazone cell pellet 24 h
SA462325Adult_Rosi_24h_1_2Glc_4SGBS mature adipocytes Rosiglitazone cell pellet 24 h
SA462326Adult_Rosi_3h_1_2Glc_2SGBS mature adipocytes Rosiglitazone cell pellet 3 h
SA462327Adult_Rosi_3h_1_2Glc_4SGBS mature adipocytes Rosiglitazone cell pellet 3 h
SA462328Adult_Rosi_3h_1_2Glc_3SGBS mature adipocytes Rosiglitazone cell pellet 3 h
SA462329Adult_Rosi_3h_1_2Glc_1SGBS mature adipocytes Rosiglitazone cell pellet 3 h
SA462330Adult_Rosi_SN_24h_1_2Glc_1SGBS mature adipocytes Rosiglitazone cell supernatant 24 h
SA462331Adult_Rosi_SN_24h_1_2Glc_2SGBS mature adipocytes Rosiglitazone cell supernatant 24 h
SA462332Adult_Rosi_SN_24h_1_2Glc_3SGBS mature adipocytes Rosiglitazone cell supernatant 24 h
SA462333Adult_Rosi_SN_24h_1_2Glc_4SGBS mature adipocytes Rosiglitazone cell supernatant 24 h
SA462334Adult_Rosi_SN_3h_1_2Glc_1SGBS mature adipocytes Rosiglitazone cell supernatant 3 h
SA462335Adult_Rosi_SN_3h_1_2Glc_2SGBS mature adipocytes Rosiglitazone cell supernatant 3 h
SA462336Adult_Rosi_SN_3h_1_2Glc_3SGBS mature adipocytes Rosiglitazone cell supernatant 3 h
SA462337Adult_Rosi_SN_3h_1_2Glc_4SGBS mature adipocytes Rosiglitazone cell supernatant 3 h
SA462338Pre_Ctrl_24h_1_2Glc_4SGBS preadipocytes Control cell pellet 24 h
SA462339Pre_Ctrl_24h_1_2Glc_3SGBS preadipocytes Control cell pellet 24 h
SA462340Pre_Ctrl_24h_1_2Glc_1SGBS preadipocytes Control cell pellet 24 h
SA462341Pre_Ctrl_24h_1_2Glc_2SGBS preadipocytes Control cell pellet 24 h
SA462342Pre_Ctrl_3h_1_2Glc_3SGBS preadipocytes Control cell pellet 3 h
SA462343Pre_Ctrl_3h_1_2Glc_2SGBS preadipocytes Control cell pellet 3 h
SA462344Pre_Ctrl_3h_1_2Glc_1SGBS preadipocytes Control cell pellet 3 h
SA462345Pre_Ctrl_3h_1_2Glc_4SGBS preadipocytes Control cell pellet 3 h
SA462346Pre_Ctrl_SN_24h_1_2Glc_1SGBS preadipocytes Control cell supernatant 24 h
SA462347Pre_Ctrl_SN_24h_1_2Glc_2SGBS preadipocytes Control cell supernatant 24 h
SA462348Pre_Ctrl_SN_24h_1_2Glc_3SGBS preadipocytes Control cell supernatant 24 h
SA462349Pre_Ctrl_SN_24h_1_2Glc_4SGBS preadipocytes Control cell supernatant 24 h
SA462350Pre_Ctrl_SN_3h_1_2Glc_2SGBS preadipocytes Control cell supernatant 3 h
SA462351Pre_Ctrl_SN_3h_1_2Glc_4SGBS preadipocytes Control cell supernatant 3 h
SA462352Pre_Ctrl_SN_3h_1_2Glc_3SGBS preadipocytes Control cell supernatant 3 h
SA462353Pre_Ctrl_SN_3h_1_2Glc_1SGBS preadipocytes Control cell supernatant 3 h
SA462354Pre_MINCH_10nM_24h_1_2Glc_2SGBS preadipocytes MINCH 10nM cell pellet 24 h
SA462355Pre_MINCH_10nM_24h_1_2Glc_4SGBS preadipocytes MINCH 10nM cell pellet 24 h
SA462356Pre_MINCH_10nM_24h_1_2Glc_3SGBS preadipocytes MINCH 10nM cell pellet 24 h
SA462357Pre_MINCH_10nM_24h_1_2Glc_1SGBS preadipocytes MINCH 10nM cell pellet 24 h
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Collection:

Collection ID:CO004151
Collection Summary:The SGBS cells were obtained from Prof. Martin Wabitsch laboratory at the University Clinic Ulm. SGBS preadipocytes were differentiated according to the standard protocol described previously (Wabitsch et al., 2001, DOI: 10.1038/sj.ijo.0801520).
Sample Type:Adipose tissue

Treatment:

Treatment ID:TR004167
Treatment Summary:SGSB preadipocytes and mature adipocytes were maintained at 37 °C, 5 % CO2, and 95 % humidity. Treatment of preadipocytes and mature adipocytes was performed as described in Study 1. For isotope labeling under insulin-stimulated conditions, cells were kept insulin-deprived for 16 hours, and the medium was replaced with 3FC medium prepared with custom-made DMEM/F12 in which glucose was replaced with the stable isotope-labeled analog [1,2 13C]glucose and conditioned according to treatment (DINCH, MINCH rosiglitazone, or control). Labeling incubation was performed for 3 and 24 hours, followed by metabolite extraction. A final concentration of 0.01% (v/v) MeOH and 0.02% (v/v) DMSO was added to all conditioned differentiation media. Continuous exposure was mimicked by replacing the cell culture medium every other day. Each treatment was performed in four biological replicates (n=4).

Sample Preparation:

Sampleprep ID:SP004164
Sampleprep Summary:Intracellular and extracellular metabolites were extracted with 1:1:1 methanol:water:chloroform. To extract the intracellular metabolites, the culture medium was removed, the cells were rinsed twice with 1 ml 0.9 % ice-cold NaCl, and the metabolism was stopped by adding MeOH (-20 °C) followed by the addition of ice-cold H2O containing 10 µM d6-glutarate in equal amounts. The cells were collected by cell lifter and chloroform was added to the lysate. After shaking for 20 minutes at 1,400 rpm and 4 °C, the extraction mixture was centrifuged at 18,000 g and 4 °C for 5 minutes. The polar upper phase was then collected and evaporated to complete dryness. For extraction of the extracellular metabolites, 300 µL of the supernatant was extracted in a 1:1 methanol:water:chloroform ratio, again with MeOH (-20 °C) containing 100 nM MEHP and ice-cold H2O containing 40 µM d6-glutarate. The following sample preparation was identical to the extraction of the intracellular metabolites. Note: After LC-MS measurement of the samples, the raw AUC isotopologue values provided here were corrected by 1.1 % of the 13C natural abundance using the R-based IsoCorrectoR tool (Heinrich et al., 2018). After correction, the relative 13C isotopolog abundances and the 13C fractional contributions from glucose and glutamine were calculated for each metabolite.

Chromatography:

Chromatography ID:CH005035
Chromatography Summary:Solvent A: 10mM tributylamine, 10mM acetic acid, 5% MeOH, 2% 2-propanol in water; Solvent B: 100% 2-propanol
Instrument Name:Agilent 1290 Infinity II
Column Name:Waters Xselect XP HSS T3 (150 x 2.1mm, 2.5um)
Column Temperature:40
Flow Gradient:0-5 min 0% B, 5-9 min 0%- 2% B, 9-9.5 min 2-6% B, 9.5-11.5 min 6% B, 11.5-12 min 6-11% B, 12-13.5 min 11% B, 13.5-15.5 min 11-28% B, 15.5-16.5 min 28-53% B, 16.5-22.5 53% B, 22.5-23 min 53-0% B, 23-33 min 0% B
Flow Rate:0-15.5 min 0.4 mL/min, 15.5-16.5 min 0.4-0.15 mL/min, 16.5-23 min 0.15 mL/min, 23-27 min 0.15-0.4 mL/min, 27-33 min 0.4 mL/min
Solvent A:93% water/5% methanol/2% isopropanol; 10mM tributylamine; 10mM acetic acid
Solvent B:100% isopropanol
Chromatography Type:Reversed phase

Analysis:

Analysis ID:AN006626
Analysis Type:MS
Chromatography ID:CH005035
Num Factors:36
Num Metabolites:123
Units:Peak AUC
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