Summary of Study ST000529

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench,, 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.


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Study IDST000529
Study TitleRegulation of Metabolism by LSR (part II)
Study TypeBroad spectrum, reverse phase LCMS metabolomics (Negative ion mode)
Study SummaryOur 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.
University of North Carolina
DepartmentSystems and Translational Sciences
LaboratorySumner Lab
Last NameSumner
First NameSusan
AddressEastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081
Submit Date2016-12-30
Num Groups4
Total Subjects20
Raw Data AvailableYes
Raw Data File Type(s)raw(Waters)
Analysis Type DetailLC-MS
Release Date2018-02-07
Release Version1
Susan Sumner Susan Sumner application/zip

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Project ID:PR000388
Project DOI:doi: 10.21228/M8WP4C
Project Title:Regulation of Metabolism by LSR
Project Type:LC-MS Lipidomics
Project Summary:Breast cancer is a global crisis, accounting for nearly one quarter of all cancers in women. In the U.S., African-American (AA) women suffer disproportionately from breast cancer mortality compared to other racial/ethnic groups. Both social (socioeconomic factors, diet, obesity) and biological hypotheses (gene polymorphisms, gene expression profiling) have been advanced to explain these discrepancies, but the differences remain poorly understood. Multiple aspects of tumor aggressiveness have been identified in the AA population, including a high proportion of basal-like tumors. Basal-like tumors are clinically the most aggressive, characterized by enhanced cancer stem cell-like features. Currently, no effective molecular therapies exist for these highly aggressive cancers and patient survival is poor. Escalating this disparity is the disease promoting effects of obesity and metabolic syndrome, which are significantly higher in AA women. Obesity and its associated inflammation have been attributed to poor patient outcomes, resistance to chemotherapeutics, and/or cancer risk. A meta-analysis of 43 studies of obesity and breast cancer revealed that obese patients were 33% more likely than non-obese patients to die of breast cancer. Yet apart from correlative studies, no reports have combined these factors with cancer disparities to ascertain their molecular interactions and physiology on breast tumorigenesis. We tested the integration these factors, with focus on a specific molecule, Lipolysis Stimulated Lipoprotein Receptor (LSR), in the promotion of aggressive cancer behaviors. LSR is a cell surface molecule that regulates post-prandial lipid uptake in the liver, is sensitive to high fat diets, and is regulated by metabolic cues, including leptin. By testing each of these factors as well as their dynamic interactions on breast cancer tumorigenesis, we will gain valuable insight into biological mechanisms that influence cancer risk/disparities, response to therapy, and ultimately patient outcome. We recently reported LSR is overexpressed in breast tumors, directs aggressive breast cancer cell behaviors including proliferation and migration, and enhances cancer stem cell-like and chemotherapeutic resistance features in breast cancer cells.
Institute:North Carolina Central University
Department:Department of Biology
Last Name:Fleming
First Name:Jodie
Address:1801 Fayetteville Street, Durham, NC 27707


Subject ID:SU000551
Subject Type:Cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Subject Comments:Cell pellets stored at -80˚C.
Cell Counts:1E 7 cells/pellet
Species Group:Human


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

mb_sample_id local_sample_id Cell Line
SA027785NEG_RP_S_33_HS#2Hs578t parental
SA027786NEG_RP_S_34_HS#2Hs578t parental
SA027787NEG_RP_S_35_HS#2Hs578t parental
SA027788NEG_RP_S_32_HSHs578t parental
SA027789NEG_RP_S_31_HS2Hs578t parental
SA027796NEG_RP_S_25_MCF7MCF7 parental
SA027797NEG_RP_S_21_MCF7-1MCF7 parental
SA027798NEG_RP_S_22_MCF7-2MCF7 parental
SA027799NEG_RP_P_29_JFtotPool2Study Pool
SA027800NEG_RP_P_31_JFtotPool4Study Pool
SA027801NEG_RP_EQ _P28_JFtotPool1-6Study Pool
SA027802NEG_RP_P_30_JFtotPool3Study Pool
Showing results 1 to 18 of 18


Collection ID:CO000545
Collection Summary:20 breast cancer cell pellets from four cell lines [MCF7 parental (LSR+), MCF7 Crisper (LSR-), Hs578t parental (LSR-) and Hs578t (LSR++)], with five replicate samples for each cell line were stored at -80˚ until sample preparation.
Sample Type:Cell pellets
Storage Conditions:-80˚C


Treatment ID:TR000565
Treatment Summary:Cultured in an obesogenic media.

Sample Preparation:

Sampleprep ID:SP000558
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 reverse phase, transferred the maximum clean volume of the upper aqueous layer to a new, pre-labeled LoBind tube then a 150 µL aliquot was transferred to a new 1.5 mL, LoBind tube for analysis. An additional 50 µL aliquot of each study sample was combined in a 2 mL LoBind tube to make a QC pool, which was vortexed for 30 sec and aliquoted into 5 Total Pool samples (150 µL each), and the remainder used for 1 Equilibrium sample (for column conditioning). Samples were placed at -80˚C for 60 mins and lyophilized to dryness. Samples were reconstituted in H2O/Methanol (95:5 v/v), vortexed at 5,000 rpm for 2 mins and centrifuged at 16,000 rcf for 4 min. 100 µL of each supernatant was transferred to pre-labeled autosampler vials and 5 µL was injected Synapt G2Si ESI-Q-TOF for data acquisition.

Combined analysis:

Analysis ID AN000807
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity
Column Waters Acquity HSS T3 (100 x 2.1mm,1.8um)
MS instrument type QTOF
MS instrument name Waters Synapt G2 Si QTOF
Units m/z


Chromatography ID:CH000582
Chromatography Summary:Reversed-Phase Gradient Seperation
Methods Filename:RTI-RCMRC-RP
Instrument Name:Waters Acquity
Column Name:Waters Acquity HSS T3 (100 x 2.1mm,1.8um)
Column Temperature:50 °C
Flow Rate:0.4 mL/min
Internal Standard:L-Tryptophan-d5
Solvent A:100% water; 0.1% formic acid
Solvent B:100% methanol; 0.1% formic acid
Analytical Time:22 min
Weak Wash Solvent Name:10% MeOH
Weak Wash Volume:1000 µL
Strong Wash Solvent Name:80% MeOH
Strong Wash Volume:1000 µL
Target Sample Temperature:8 °C
Sample Loop Size:10 µL
Sample Syringe Size:100 µL
Randomization Order:Yes
Chromatography Type:Reversed phase


MS ID:MS000714
Analysis ID:AN000807
Instrument Name:Waters Synapt G2 Si QTOF
Instrument Type:QTOF
Capillary Temperature:110 °C
Capillary Voltage:1.0 kV
Collision Energy:4
Fragmentation Method:CID
Helium Flow:180
Ionization Potential:5 ppm
Source Temperature:100 °C
Desolvation Gas Flow:400 L/Hr
Desolvation Temperature:400 °C
Resolution Setting:18000
Scan Range Moverz:50-1000 m/z
Scanning Cycle:1.0 s
Tube Lens Voltage:74