Summary of Study ST000955

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 PR000656. The data can be accessed directly via it's Project DOI: 10.21228/M8W39D 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 IDST000955
Study TitleEffects of ODC inhibition on T cell metabolism
Study Typedrug treatment at two time
Study SummaryIsolated T cells were activated for 48-72 hours in the presence and absence of 5mM DFMO, an ODC inhibitor
Institute
University of Florida
DepartmentSECIM
Last NameHesterberg
First NameRebecca
Address12902 Magnolia Dr
Emailrebecca.hesterberg@moffitt.org
Phone813-270-9181
Submit Date2018-04-14
Num Groups4
Total Subjects16
Study CommentsOT-1 mice express a transgenic T cell receptor in all CD8+ T cells that binds with high affinity to a peptide derived from the protein ovalbumin (SIINFEKL).
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2019-05-15
Release Version1
Rebecca Hesterberg Rebecca Hesterberg
https://dx.doi.org/10.21228/M8W39D
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR000656
Project DOI:doi: 10.21228/M8W39D
Project Title:Cereblon suppression offers novel approach for anti-tumor immunity in melanoma
Project Type:Exploratory
Project Summary:The functional suppression of T cells in the tumor are intimately linked to impared cellular metabolism. Glucose acts as the primary fuel source for the generation of ATP in activated T cells and both oxidative phosphorylation and glycolysis are critical for full effector functions and tumor erradication. We have identified a checkpoint protein that regulates the metabolic suppression in the tumor. Identification of the precise function of different metabolic events regulated by this checkpoint protein will be informative. This project will define the role of a glutamine/arginine mediated pathway in CD8+ T-cells.
Institute:Moffitt Cancer Center
Department:Immunology
Laboratory:Burnette
Last Name:Burnette
First Name:Pearlie
Address:12902 Magnolia Drive, Tampa, FL 33612
Email:Pearlie.Burnette@moffitt.org
Phone:813-270-9181
Funding Source:Moffitt Cancer Center

Subject:

Subject ID:SU000994
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57Bl6 OT-1
Age Or Age Range:6-8 weeks
Animal Animal Supplier:Breeding at Moffitt for several generations, originally from Jax
Animal Light Cycle:12 hour

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Time activated with peptide DFMO treatment (mM)
SA057203WT1_48h_0mM_148 -
SA057204WT3_48h_0mM_348 -
SA057205WT3_48h_0mM_248 -
SA057206WT7_48h_0mM_248 -
SA057207WT8_48h_0mM_148 -
SA057208WT8_48h_0mM_348 -
SA057209WT8_48h_0mM_248 -
SA057210WT3_48h_0mM_148 -
SA057211WT7_48h_0mM_348 -
SA057212WT7_48h_0mM_148 -
SA057213WT1_48h_0mM_348 -
SA057214WT1_48h_0mM_248 -
SA057215WT1_48h_5mM_248 5
SA057216WT7_48h_5mM_148 5
SA057217WT1_48h_5mM_148 5
SA057218WT7_48h_5mM_248 5
SA057219WT8_48h_5mM_148 5
SA057220WT8_48h_5mM_348 5
SA057221WT8_48h_5mM_248 5
SA057222WT1_48h_5mM_348 5
SA057223WT7_48h_5mM_348 5
SA057224WT3_48h_5mM_248 5
SA057225WT3_48h_5mM_148 5
SA057226WT3_48h_5mM_348 5
SA057227WT8_72h_0mM_372 -
SA057228WT7_72h_0mM_372 -
SA057229WT8_72h_0mM_272 -
SA057230WT3_72h_0mM_172 -
SA057231WT8_72h_0mM_172 -
SA057232WT1_72h_0mM_372 -
SA057233WT7_72h_0mM_272 -
SA057234WT1_72h_0mM_172 -
SA057235WT1_72h_0mM_272 -
SA057236WT7_72h_0mM_172 -
SA057237WT3_72h_0mM_372 -
SA057238WT3_72h_0mM_272 -
SA057239WT1_72h_5mM_272 5
SA057240WT8_72h_5mM_172 5
SA057241WT8_72h_5mM_272 5
SA057242WT8_72h_5mM_372 5
SA057243WT7_72h_5mM_372 5
SA057244WT3_72h_5mM_172 5
SA057245WT3_72h_5mM_372 5
SA057246WT1_72h_5mM_372 5
SA057247WT7_72h_5mM_172 5
SA057248WT3_72h_5mM_272 5
SA057249WT7_72h_5mM_272 5
SA057250WT1_72h_5mM_172 5
Showing results 1 to 48 of 48

Collection:

Collection ID:CO000988
Collection Summary:At 48 or 72 hours cells in culture medium were centrifuged on a Ficoll gradient to enrich viable cells. Cells were washed 1x with PBS, counted, and frozen as cell pellets at -80.
Sample Type:Cultured cells

Treatment:

Treatment ID:TR001008
Treatment Summary:Splenic CD8+ T cells were isolated from 4 OT-1 mice (WT1, 3, 7, 8) by negative immunomagnetic selection. T cells stimulated with ova-derived peptide (SIINFEKL) that binds with high affinity to the OT-1 transgenic TCR and 10ng/mL IL-2 with and without 5mM DFMO for 48-72 hours. Cell stoarge: 37 degrees, Cell growth container: 24 well plate, cell media: RPMI1640 with glutamine. Added components were: NEAA, Sodium Pyruvate, Fetal Bovine Serum, Fungizone, Gentimycin, Pen/strep.

Sample Preparation:

Sampleprep ID:SP001001
Sampleprep Summary:None
Sampleprep Protocol Filename:GMetabolomics_LCMS_Protocol_092117.pdf
Appendix_A_Internal_Standard_Prep_GLCMS.pdf

Combined analysis:

Analysis ID AN001566 AN001567
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo Dionex Ultimate 3000 Thermo Dionex Ultimate 3000
Column ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) ACE Excel 2 C18-PFP (100 x 2.1mm, 2um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive Orbitrap Thermo Q Exactive Orbitrap
Ion Mode POSITIVE NEGATIVE
Units peak height peak height

Chromatography:

Chromatography ID:CH001098
Methods Filename:GMetabolomics_LCMS_Protocol_092117.pdf
Instrument Name:Thermo Dionex Ultimate 3000
Column Name:ACE Excel 2 C18-PFP (100 x 2.1mm, 2um)
Flow Rate:350 ul/min
Solvent A:100% water; 0.1% formic acid
Solvent B:100% acetonitrile
Chromatography Type:Reversed phase

MS:

MS ID:MS001444
Analysis ID:AN001566
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS001445
Analysis ID:AN001567
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
Ion Mode:NEGATIVE
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