{ "METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000955","ANALYSIS_ID":"AN001567","VERSION":"1","CREATED_ON":"April 19, 2018, 11:02 am"}, "PROJECT":{"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"}, "STUDY":{"STUDY_TITLE":"Effects of ODC inhibition on T cell metabolism","STUDY_TYPE":"drug treatment at two time","STUDY_SUMMARY":"Isolated T cells were activated for 48-72 hours in the presence and absence of 5mM DFMO, an ODC inhibitor","INSTITUTE":"University of Florida","DEPARTMENT":"SECIM","LAST_NAME":"Hesterberg","FIRST_NAME":"Rebecca","ADDRESS":"12902 Magnolia Dr","EMAIL":"rebecca.hesterberg@moffitt.org","PHONE":"813-270-9181","NUM_GROUPS":"4","TOTAL_SUBJECTS":"16","STUDY_COMMENTS":"OT-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)."}, "SUBJECT":{"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"}, "SUBJECT_SAMPLE_FACTORS":[ { "Subject ID":"WT1", "Sample ID":"WT1_48h_0mM_1", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT1", "Sample ID":"WT1_48h_0mM_2", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT1", "Sample ID":"WT1_48h_0mM_3", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT1", "Sample ID":"WT1_48h_5mM_1", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT1", "Sample ID":"WT1_48h_5mM_2", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT1", "Sample ID":"WT1_48h_5mM_3", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT1", "Sample ID":"WT1_72h_0mM_1", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT1", "Sample ID":"WT1_72h_0mM_2", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT1", "Sample ID":"WT1_72h_0mM_3", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT1", "Sample ID":"WT1_72h_5mM_1", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT1", "Sample ID":"WT1_72h_5mM_2", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT1", "Sample ID":"WT1_72h_5mM_3", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT3", "Sample ID":"WT3_48h_0mM_1", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT3", "Sample ID":"WT3_48h_0mM_2", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT3", "Sample ID":"WT3_48h_0mM_3", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT3", "Sample ID":"WT3_48h_5mM_1", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT3", "Sample ID":"WT3_48h_5mM_2", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT3", "Sample ID":"WT3_48h_5mM_3", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT3", "Sample ID":"WT3_72h_0mM_1", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT3", "Sample ID":"WT3_72h_0mM_2", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT3", "Sample ID":"WT3_72h_0mM_3", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT3", "Sample ID":"WT3_72h_5mM_1", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT3", "Sample ID":"WT3_72h_5mM_2", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT3", "Sample ID":"WT3_72h_5mM_3", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT7", "Sample ID":"WT7_48h_0mM_1", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT7", "Sample ID":"WT7_48h_0mM_2", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT7", "Sample ID":"WT7_48h_0mM_3", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT7", "Sample ID":"WT7_48h_5mM_1", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT7", "Sample ID":"WT7_48h_5mM_2", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT7", "Sample ID":"WT7_48h_5mM_3", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT7", "Sample ID":"WT7_72h_0mM_1", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT7", "Sample ID":"WT7_72h_0mM_2", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT7", "Sample ID":"WT7_72h_0mM_3", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT7", "Sample ID":"WT7_72h_5mM_1", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT7", "Sample ID":"WT7_72h_5mM_2", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT7", "Sample ID":"WT7_72h_5mM_3", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT8", "Sample ID":"WT8_48h_0mM_1", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT8", "Sample ID":"WT8_48h_0mM_2", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT8", "Sample ID":"WT8_48h_0mM_3", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT8", "Sample ID":"WT8_48h_5mM_1", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT8", "Sample ID":"WT8_48h_5mM_2", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT8", "Sample ID":"WT8_48h_5mM_3", "Factors":{"Time activated with peptide":"48","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT8", "Sample ID":"WT8_72h_0mM_1", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT8", "Sample ID":"WT8_72h_0mM_2", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT8", "Sample ID":"WT8_72h_0mM_3", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"-"} }, { "Subject ID":"WT8", "Sample ID":"WT8_72h_5mM_1", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT8", "Sample ID":"WT8_72h_5mM_2", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} }, { "Subject ID":"WT8", "Sample ID":"WT8_72h_5mM_3", "Factors":{"Time activated with peptide":"72","DFMO treatment (mM)":"5"} } ], "COLLECTION":{"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_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."}, "SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"None","SAMPLEPREP_PROTOCOL_FILENAME":"GMetabolomics_LCMS_Protocol_092117.pdf;Appendix_A_Internal_Standard_Prep_GLCMS.pdf"}, "CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"ACE Excel 2 C18-PFP (100 x 2.1mm, 2um)","FLOW_RATE":"350 ul/min","METHODS_FILENAME":"GMetabolomics_LCMS_Protocol_092117.pdf","SOLVENT_A":"0.1% Formic Acid in Water","SOLVENT_B":"Acetonitrile"}, "ANALYSIS":{"ANALYSIS_TYPE":"MS"}, "MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_RESULTS_FILE":"ST000955_AN001567_Results.txt UNITS:peak height"}, "MS_METABOLITE_DATA":{ "Units":"peak height", 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