Summary of Study ST000528

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 PR000388. The data can be accessed directly via it's Project DOI: 10.21228/M8WP4C 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.

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Study ID	ST000528
Study Title	Regulation of Metabolism by LSR
Study Type	LC-MS lipidomics
Study Summary	Our aim is to identify the LSR-driven metabolomics profile of breast cancer cells in lean and obesogenic environments. Breast cancer cell models with high or undetectable levels of LSR, including drug resistance models, were cultured in lean and obesogenic environments and comprehensive metabolomics profiling, including lipidomics-focused sub-analyses were performed. The metabolomics analyses using both approaches will help us determine if LSR enhances aggressive breast cancer phenotypes via modulation of cellular bioenergetic metabolism, ultimately contributing to poor patient outcome.
Institute	University of North Carolina
Department	Systems and Translational Sciences
Laboratory	Sumner Lab
Last Name	Sumner
First Name	Susan
Address	Eastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081
Email	susan_sumner @unc.edu
Phone	704-250-5066
Submit Date	2016-12-29
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2018-02-07
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP000557
Sampleprep Summary:	Cell pellets were resuspended in volume of ice cold Lipid Extractions Solvent (50 µg/mL) based on biomass. Contents were transferred to new, pre-labeled 2.0 mL LoBind tube with 10-15 ceramic beads, and homogenized using 2 pulses at 2,000 rpm for 30 sec on a MagNA Lyser. Volume of HPLC-grade water (with 0.02 mg/mL Tryptophan-d5 in water) was added based on biomass. Samples were allowed to sit at room temperature for 10 minutes, then centrifuged at 16,000 rcf for 10 min at 10˚C. For lipidomics, transferred the maximum clean volume of the lower lipid-rich DCM layer to a new, pre-labeled LoBind tube and a 375 µL aliquot was then transferred to a new 1.5 mL, LoBind tube for analysis. An additional 125 µL aliquot of each study sample was transferred to a 7.0 mL glass vial to make a QC pool, which was vortexed for 30 sec and aliquoted into 5 Total Pool samples (375 µL each), and the remaining was used for 1 Equilibrium sample (column conditioning). Samples were frozen at -80˚C for 60 mins and lyophilized to dryness. Samples were reconstituted in ACN/IPA/H2O (65:30:5 v/v/v), vortexed at 4,000 rpm for 2 mins and centrifuged at 16,000 rcf for 4 min. 225 µL of the supernatant was transferred to pre-labeled autosampler vials and 10 µL was injected into OrbiTrap Velos.

Sampleprep ID:

SP000557

Sampleprep Summary:

Cell pellets were resuspended in volume of ice cold Lipid Extractions Solvent (50 µg/mL) based on biomass. Contents were transferred to new, pre-labeled 2.0 mL LoBind tube with 10-15 ceramic beads, and homogenized using 2 pulses at 2,000 rpm for 30 sec on a MagNA Lyser. Volume of HPLC-grade water (with 0.02 mg/mL Tryptophan-d5 in water) was added based on biomass. Samples were allowed to sit at room temperature for 10 minutes, then centrifuged at 16,000 rcf for 10 min at 10˚C. For lipidomics, transferred the maximum clean volume of the lower lipid-rich DCM layer to a new, pre-labeled LoBind tube and a 375 µL aliquot was then transferred to a new 1.5 mL, LoBind tube for analysis. An additional 125 µL aliquot of each study sample was transferred to a 7.0 mL glass vial to make a QC pool, which was vortexed for 30 sec and aliquoted into 5 Total Pool samples (375 µL each), and the remaining was used for 1 Equilibrium sample (column conditioning). Samples were frozen at -80˚C for 60 mins and lyophilized to dryness. Samples were reconstituted in ACN/IPA/H2O (65:30:5 v/v/v), vortexed at 4,000 rpm for 2 mins and centrifuged at 16,000 rcf for 4 min. 225 µL of the supernatant was transferred to pre-labeled autosampler vials and 10 µL was injected into OrbiTrap Velos.