Summary of Study ST003903
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 PR002443. The data can be accessed directly via it's Project DOI: 10.21228/M8VG1V This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
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.
| Study ID | ST003903 |
| Study Title | Hypertonic stress promotes intracellular lipid accumulation via glutamine anaplerosis |
| Study Type | Untargeted lipidomics |
| Study Summary | In this work, we describe that acute hypertonic stress triggers the upregulation of metabolic pathways that promote glutamine-dependent lipid accumulation associated with decreased oxygen consumption thus increasing potential water storage. Motive: We sought to understand the lipidomic profile of cells exposed to hypertonic stress, as hypertonic stress is known to regulate lipid metabolism. Methods: We treated mouse kidney inner medullary collecting duct cells with 100 mmol NaCl for 12 hours and then collected cell lysates for evaluation of the lipidomic profile via LC-MS/MS Samples: Control (n=4) – no exposure to 100 mmol NaCl – treated with DMEM low glucose no FBS or antibiotic/antimycotic. Salt (n=5) – Exposed to 100 mmol NaCl – treated with DMEM low glucose with no FBS or antibiotic/antimycotic. Results: We found that long chain triglycerides and cardiolipins are increased after exposure to NaCl. |
| Institute | Vanderbilt University |
| Department | Chemistry |
| Laboratory | Center for Innovative Technology |
| Last Name | Leaptrot |
| First Name | Katrina |
| Address | 7300 Stevenson Center Lane |
| katrina.l.leaptrot@vanderbilt.edu | |
| Phone | 6158228422 |
| Submit Date | 2025-04-30 |
| Num Groups | 2 |
| Total Subjects | 9 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, d |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-10-30 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002443 |
| Project DOI: | doi: 10.21228/M8VG1V |
| Project Title: | Hypertonic stress promotes intracellular lipid accumulation via glutamine anaplerosis |
| Project Type: | Untargeted lipidomics |
| Project Summary: | In this work, we describe that acute hypertonic stress triggers the upregulation of metabolic pathways that promote glutamine-dependent lipid accumulation associated with decreased oxygen consumption thus increasing potential water storage. Motive: We sought to understand the lipidomic profile of cells exposed to hypertonic stress, as hypertonic stress is known to regulate lipid metabolism. Methods: We treated mouse kidney inner medullary collecting duct cells with 100 mmol NaCl for 12 hours and then collected cell lysates for evaluation of the lipidomic profile via LC-MS/MS Samples: Control (n=4) – no exposure to 100 mmol NaCl – treated with DMEM low glucose no FBS or antibiotic/antimycotic Salt (n=5) – Exposed to 100 mmol NaCl – treated with DMEM low glucose with no FBS or antibiotic/antimycotic. Results: We found that long chain triglycerides and cardiolipins are increased after exposure to NaCl. |
| Institute: | Vanderbilt University |
| Department: | Chemistry |
| Laboratory: | Center for Innovative Technology |
| Last Name: | Leaptrot |
| First Name: | Katrina |
| Address: | 7300 Stevenson Center Lane |
| Email: | katrina.l.leaptrot@vanderbilt.edu |
| Phone: | 6158228422 |
Subject:
| Subject ID: | SU004038 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Cell Strain Details: | kidney inner medullary collecting duct cells |
Factors:
Subject type: Cultured cells; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment |
|---|---|---|
| SA429748 | Ctrl 1_neg | Control |
| SA429749 | Ctrl 4_pos | Control |
| SA429750 | Ctrl 1_pos | Control |
| SA429751 | Ctrl 3_pos | Control |
| SA429752 | Ctrl 3_neg | Control |
| SA429753 | Ctrl 2_neg | Control |
| SA429754 | Ctrl 4_neg | Control |
| SA429755 | Ctrl 2_pos | Control |
| SA429756 | NaCl 3_neg | NaCl |
| SA429757 | NaCl 5_neg | NaCl |
| SA429758 | NaCl 2_neg | NaCl |
| SA429759 | NaCl 1_neg | NaCl |
| SA429760 | NaCl 4_neg | NaCl |
| SA429761 | NaCl 5_pos | NaCl |
| SA429762 | NaCl 3_pos | NaCl |
| SA429763 | NaCl 4_pos | NaCl |
| SA429764 | NaCl 1_pos | NaCl |
| SA429765 | NaCl 2_pos | NaCl |
| Showing results 1 to 18 of 18 |
Collection:
| Collection ID: | CO004031 |
| Collection Summary: | Inner medullary collecting duct cells were cultured in standard low glucose DMEM supplemented with 10% FBS, antibiotic and antimycotic. Cells were plated in 10 cm plates and grown to 90% confluency. Medium was the changed to DMEM low glucose with or without 100 mmol NaCl added. Cells were incubated overnight and collected for downstream analysis. |
| Sample Type: | Kidney |
Treatment:
| Treatment ID: | TR004047 |
| Treatment Summary: | Treatment was DMEM low glucose with or without 100 mmol NaCl. No other additives. |
Sample Preparation:
| Sampleprep ID: | SP004044 |
| Sampleprep Summary: | Frozen samples were thawed on ice and lysed in ice-cold lysis buffer (1:1:2, acetonitrile: methanol: ammonium bicarbonate 0.1M, pH 8.0) to an equal cell density per sample, followed by probe tip sonication with 10 pulses at 30% power. Samples were normalized by total protein amount (150 µg) based on bicinchoninic acid (BCA) assay and further mixed with 800 µL of cold MeOH, vortexed for 30 seconds and incubated overnight at -80°C for protein precipitation. Following incubation, samples were centrifuged for 10 min at 15,000 rpm at 4°C and the supernatant was transferred to a new labeled tube and dried down using a cold vacuum centrifuge. Samples were reconstituted in 100 µL H2O, 100 µL MeOH, and 10 µL of SPLASH LIPIDOMIX with vortex mixing after each addition. Samples were incubated at room temperature for 10 min followed by liquid-liquid extraction. For liquid-liquid extraction (LLE), 600 µL MTBE was added with vortex mixing for 30 seconds followed by incubation on ice for 10 min and centrifugation at 15,000 rpm for 15 minutes at 4°C. An upper (hydrophobic) fraction was transferred and dried down using cold vacuum centrifuge and stored at -80°C for further lipidomic studies. Prior to mass spectrometry analysis, individual hydrophobic extracts were reconstituted in 80 µL methanol:chloroform (9:1, v:v) containing exogenous lipid standards to assess instrument variability. A pooled quality control (QC) sample was prepared by pooling equal volumes (30 μL) from each individual sample following reconstitution. |
| Sampleprep Protocol Filename: | Global_Untargeted_Lipidomics_Methods.pdf |
Chromatography:
| Chromatography ID: | CH004859 |
| Methods Filename: | Global_Untargeted_Lipidomics_Methods.pdf |
| Instrument Name: | Agilent 1290 Infinity |
| Column Name: | Thermo Hypersil GOLD aQ (100 x 2.1mm,1.9um) |
| Column Temperature: | 40 |
| Flow Gradient: | 0.00 min 70% B; 0.50 min 30% B; 2.00 min 70% B; 15.00 min 100% B; 21.00 min 100% B; 22.00 min 10% B; 24.00 min 10% B; 25.00 min 30% B; 30 min 70% B |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 100% water; 10 mM ammonium acetate; 0.1% formic acid |
| Solvent B: | 60% acetonitrile/36% isopropanol/4% water; 10 mM ammonium acetate; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
Analysis:
| Analysis ID: | AN006407 |
| Analysis Type: | MS |
| Analysis Protocol File: | Global_Untargeted_Lipidomics_Methods.pdf |
| Chromatography ID: | CH004859 |
| Has Mz: | 1 |
| Has Rt: | 1 |
| Rt Units: | Minutes |
| Results File: | ST003903_AN006407_Results.txt |
| Units: | Abundance |
| Analysis ID: | AN006408 |
| Analysis Type: | MS |
| Analysis Protocol File: | Global_Untargeted_Lipidomics_Methods.pdf |
| Chromatography ID: | CH004859 |
| Has Mz: | 1 |
| Has Rt: | 1 |
| Rt Units: | Minutes |
| Results File: | ST003903_AN006408_Results.txt |
| Units: | Abundance |
