Summary of Study ST001983
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 PR001259. The data can be accessed directly via it's Project DOI: 10.21228/M8ZX2N This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
Study ID | ST001983 |
Study Title | Metabolomic Fingerprinting of Human High Grade Serous Ovarian Carcinoma Cell Lines |
Study Summary | Focusing on defining the metabolomic basis of intratumoral heterogeneity in ovarian cancer, the metabolic diversity of a panel of high grade serous ovarian carcinoma (HGSOC) cell-lines we investigated using a metabolomics platform that interrogate 731 compounds. Metabolic fingerprinting followed by 2-dimensional and 3-dimensional principal component analysis defined the heterogeneity of the HGSOC cells and clustered them into five distinct metabolic groups. An overall increase in the metabolites associated with aerobic glycolysis and phospholipid metabolism were observed in the majority of the cancer cells. A preponderant increase in the levels of metabolites involved in trans-sulfuration and glutathione synthesis was also observed. Subsets of HGSOC cells showed an increase in the levels of 5-Hydroxytryptamine, gamma-aminobutyric acid, or glutamate, pointing to their potential role as oncometabolites. In summary, our results identify increased glycolysis, phospholipid metabolism and amino acid metabolism with the resultant increase in the levels of 5-Hydoxytryptamine, GABA, and Glutamate as metabolomic correlates underlying the heterogeneity of ovarian cancer cell lines. |
Institute | University of Oklahoma Health Sciences Center |
Department | Cell Biology |
Laboratory | Danny N Dhanasekaran |
Last Name | Jayaraman |
First Name | Muralidharan |
Address | 975 NE 10th street, BRC1470, Oklahoma City, Oklahoma, 73104, USA |
Muralidharan-Jayaraman@ouhsc.edu | |
Phone | 4052718001 x 30492 |
Submit Date | 2021-09-20 |
Num Groups | 2 |
Total Subjects | 30 |
Num Females | 15 |
Study Comments | Ovarian cancer cell lines |
Analysis Type Detail | LC-MS |
Release Date | 2021-12-01 |
Release Version | 1 |
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Combined analysis:
Analysis ID | AN003234 |
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Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Waters ACQUITY ultra-performance liquid chromatography (UPLC) |
Column | Waters UPLC BEH C18 (100 x 2.1mm,1.7um) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Q Exactive HF hybrid Orbitrap |
Ion Mode | UNSPECIFIED |
Units | ratio |
MS:
MS ID: | MS003008 |
Analysis ID: | AN003234 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | All methods utilized a Waters ACQUITY ultra-performance liquid chromatography (UPLC) and a Thermo Scientific Q-Exactive high resolution/accurate mass spectrometer interfaced with a heated electrospray ionization (HESI-II) source and Orbitrap mass analyzer operated at 35,000 mass resolution. The sample extract was dried then reconstituted in solvents compatible to each of the four methods. Each reconstitution solvent contained a series of standards at fixed concentrations to ensure injection and chromatographic consistency. One aliquot was analyzed using acidic positive ion conditions, chromatographically optimized for more hydrophilic compounds. In this method, the extract was gradient eluted from a C18 column (Waters UPLC BEH C18-2.1x100 mm, 1.7 µm) using water and methanol, containing 0.05% perfluoropentanoic acid (PFPA) and 0.1% formic acid (FA). Another aliquot was also analyzed using acidic positive ion conditions, however it was chromatographically optimized for more hydrophobic compounds. In this method, the extract was gradient eluted from the same afore mentioned C18 column using methanol, acetonitrile, water, 0.05% PFPA and 0.01% FA and was operated at an overall higher organic content. Another aliquot was analyzed using basic negative ion optimized conditions using a separate dedicated C18 column. The basic extracts were gradient eluted from the column using methanol and water, however with 6.5mM Ammonium Bicarbonate at pH 8. The fourth aliquot was analyzed via negative ionization following elution from a HILIC column (Waters UPLC BEH Amide 2.1x150 mm, 1.7 µm) using a gradient consisting of water and acetonitrile with 10mM Ammonium Formate, pH 10.8. The MS analysis alternated between MS and data-dependent MSn scans using dynamic exclusion. The scan range varied slighted between methods but covered 70-1000 m/z. Raw data files are archived and extracted using proprietary, Metabolon LIMS data structures and analysis software (Metabolon Inc, Durham). Briefly, raw data was extracted, peak-identified and QC processed using Metabolon’s hardware and software. Biochemical identifications are based on three criteria: retention index within a narrow RI window of the proposed identification, accurate mass match to the library +/- 10 ppm, and the MS/MS forward and reverse scores between the experimental data and authentic standards. |
Ion Mode: | UNSPECIFIED |