{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000454","ANALYSIS_ID":"AN000711","VERSION":"1","CREATED_ON":"September 7, 2016, 2:21 pm"},

"PROJECT":{"PROJECT_TITLE":"Utilizing Metabolomics to Understand Novel Anti-Desmoid Tumor Drugs","PROJECT_SUMMARY":"Desmoid tumors (DT) are locally invasive soft tissue growths with no directed therapies currently. While two genes (β-catenin and adenomatous polyposis coli) have been found in patients who develop desmoids, it is unclear how these mutations and other downstream mechanisms lead to desmoid tumorigenesis. Extensive research has been explored in the molecular biology of desmoids; however, the use of metabolomics to understand the how the low molecular weight complements of cells, tissues, and biological fluids are perturbed by this highly localized disease. Additionally, the Desmoid Collaboration for a Cure has identified 45 active drugs against primary cell lines. It is unclear how these therapies perturb the metabolome, outside the Wnt and notch pathways.","INSTITUTE":"RTI International","LAST_NAME":"Mercier","FIRST_NAME":"Kelly","ADDRESS":"3040 E. Cornwallis Road","EMAIL":"kmercier@rti.org","PHONE":"919-541-6396"},

"STUDY":{"STUDY_TITLE":"Utilizing Metabolomics to Understand Novel Anti-Desmoid Tumor Drugs","STUDY_SUMMARY":"This pilot study will use broad spectrum metabolomics to study the tumorigenesis process of fibroblasts to desmoids by investigating paired desmoid and fibroblast cell lines, in addition to unaffected fibroblast cells. Additionally, this pilot study will explore the effects of two of the active drugs identified on the desmoid and fibroblast cells.","INSTITUTE":"RTI International","LAST_NAME":"Mercier","FIRST_NAME":"Kelly","ADDRESS":"3040 E. Cornwallis Road","EMAIL":"kmercier@rti.org","PHONE":"919-541-6396"},

"SUBJECT":{"SUBJECT_TYPE":"Human Cells","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"CP_1",
"Sample ID":"CP_1_r1",
"Factors":{"Treatment":"Total Pool","Growth_type":"Total Pool"},
"Additional sample data":{"Patient":"Total Pool"}
},
{
"Subject ID":"CP_1",
"Sample ID":"CP_1_r2",
"Factors":{"Treatment":"Total Pool","Growth_type":"Total Pool"},
"Additional sample data":{"Patient":"Total Pool"}
},
{
"Subject ID":"CP_1",
"Sample ID":"CP_1_r3",
"Factors":{"Treatment":"Total Pool","Growth_type":"Total Pool"},
"Additional sample data":{"Patient":"Total Pool"}
},
{
"Subject ID":"CP_2",
"Sample ID":"CP_2_r4",
"Factors":{"Treatment":"Total Pool","Growth_type":"Total Pool"},
"Additional sample data":{"Patient":"Total Pool"}
},
{
"Subject ID":"CP_2",
"Sample ID":"CP_2_r5",
"Factors":{"Treatment":"Total Pool","Growth_type":"Total Pool"},
"Additional sample data":{"Patient":"Total Pool"}
},
{
"Subject ID":"CP_2",
"Sample ID":"CP_2_r6",
"Factors":{"Treatment":"Total Pool","Growth_type":"Total Pool"},
"Additional sample data":{"Patient":"Total Pool"}
},
{
"Subject ID":"CP_3",
"Sample ID":"CP_3_r1",
"Factors":{"Treatment":"Total Pool","Growth_type":"Total Pool"},
"Additional sample data":{"Patient":"Total Pool"}
},
{
"Subject ID":"CP_3",
"Sample ID":"CP_3_r2",
"Factors":{"Treatment":"Total Pool","Growth_type":"Total Pool"},
"Additional sample data":{"Patient":"Total Pool"}
},
{
"Subject ID":"CP_3",
"Sample ID":"CP_3_r3",
"Factors":{"Treatment":"Total Pool","Growth_type":"Total Pool"},
"Additional sample data":{"Patient":"Total Pool"}
},
{
"Subject ID":"C_1",
"Sample ID":"C_1_r4",
"Factors":{"Treatment":"DMSO","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_1",
"Sample ID":"C_1_r5",
"Factors":{"Treatment":"DMSO","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_1",
"Sample ID":"C_1_r6",
"Factors":{"Treatment":"DMSO","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_10",
"Sample ID":"C_10_r1",
"Factors":{"Treatment":"H2O","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_10",
"Sample ID":"C_10_r2",
"Factors":{"Treatment":"H2O","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_10",
"Sample ID":"C_10_r3",
"Factors":{"Treatment":"H2O","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_11",
"Sample ID":"C_11_r1",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_11",
"Sample ID":"C_11_r2",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_11",
"Sample ID":"C_11_r3",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_12",
"Sample ID":"C_12_r4",
"Factors":{"Treatment":"H2O","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_12",
"Sample ID":"C_12_r5",
"Factors":{"Treatment":"H2O","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_12",
"Sample ID":"C_12_r6",
"Factors":{"Treatment":"H2O","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_13",
"Sample ID":"C_13_r1",
"Factors":{"Treatment":"DMSO","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_13",
"Sample ID":"C_13_r2",
"Factors":{"Treatment":"DMSO","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_13",
"Sample ID":"C_13_r3",
"Factors":{"Treatment":"DMSO","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_14",
"Sample ID":"C_14_r1",
"Factors":{"Treatment":"Das.","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_14",
"Sample ID":"C_14_r2",
"Factors":{"Treatment":"Das.","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_14",
"Sample ID":"C_14_r3",
"Factors":{"Treatment":"Das.","Growth_type":"Normal"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_15",
"Sample ID":"C_15_r1",
"Factors":{"Treatment":"DMSO","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_15",
"Sample ID":"C_15_r2",
"Factors":{"Treatment":"DMSO","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_15",
"Sample ID":"C_15_r3",
"Factors":{"Treatment":"DMSO","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_16",
"Sample ID":"C_16_r1",
"Factors":{"Treatment":"Das.","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_16",
"Sample ID":"C_16_r2",
"Factors":{"Treatment":"Das.","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_16",
"Sample ID":"C_16_r3",
"Factors":{"Treatment":"Das.","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_17",
"Sample ID":"C_17_r4",
"Factors":{"Treatment":"DMSO","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_17",
"Sample ID":"C_17_r5",
"Factors":{"Treatment":"DMSO","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_17",
"Sample ID":"C_17_r6",
"Factors":{"Treatment":"DMSO","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_18",
"Sample ID":"C_18_r1",
"Factors":{"Treatment":"Das.","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_18",
"Sample ID":"C_18_r4",
"Factors":{"Treatment":"Das.","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_18",
"Sample ID":"C_18_r6",
"Factors":{"Treatment":"Das.","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_19",
"Sample ID":"C_19_r4",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_19",
"Sample ID":"C_19_r5",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_19",
"Sample ID":"C_19_r6",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_2",
"Sample ID":"C_2_r1",
"Factors":{"Treatment":"Das.","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_2",
"Sample ID":"C_2_r2",
"Factors":{"Treatment":"Das.","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_2",
"Sample ID":"C_2_r3",
"Factors":{"Treatment":"Das.","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_20",
"Sample ID":"C_20_r4",
"Factors":{"Treatment":"H2O","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_20",
"Sample ID":"C_20_r5",
"Factors":{"Treatment":"H2O","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_20",
"Sample ID":"C_20_r6",
"Factors":{"Treatment":"H2O","Growth_type":"Unaffected"},
"Additional sample data":{"Patient":"CRL7518"}
},
{
"Subject ID":"C_3",
"Sample ID":"C_3_r1",
"Factors":{"Treatment":"DMSO","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_3",
"Sample ID":"C_3_r2",
"Factors":{"Treatment":"DMSO","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_3",
"Sample ID":"C_3_r3",
"Factors":{"Treatment":"DMSO","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_4",
"Sample ID":"C_4_r1",
"Factors":{"Treatment":"Das.","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_4",
"Sample ID":"C_4_r2",
"Factors":{"Treatment":"Das.","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_4",
"Sample ID":"C_4_r3",
"Factors":{"Treatment":"Das.","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_5",
"Sample ID":"C_5_r1",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_5",
"Sample ID":"C_5_r2",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_5",
"Sample ID":"C_5_r3",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_6",
"Sample ID":"C_6_r4",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_6",
"Sample ID":"C_6_r5",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_6",
"Sample ID":"C_6_r6",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_7",
"Sample ID":"C_7_r1",
"Factors":{"Treatment":"H2O","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_7",
"Sample ID":"C_7_r2",
"Factors":{"Treatment":"H2O","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_7",
"Sample ID":"C_7_r3",
"Factors":{"Treatment":"H2O","Growth_type":"Normal"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_8",
"Sample ID":"C_8_r1",
"Factors":{"Treatment":"H2O","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_8",
"Sample ID":"C_8_r2",
"Factors":{"Treatment":"H2O","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_8",
"Sample ID":"C_8_r3",
"Factors":{"Treatment":"H2O","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"219"}
},
{
"Subject ID":"C_9",
"Sample ID":"C_9_r1",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_9",
"Sample ID":"C_9_r2",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
},
{
"Subject ID":"C_9",
"Sample ID":"C_9_r3",
"Factors":{"Treatment":"FAK Inh. 14","Growth_type":"Tumor"},
"Additional sample data":{"Patient":"141"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"An aliquot of the media following treatment was collected, and the remainder of the media was aspirated. The cells were washed with PBS twice, and quenched with 8mL ice-cold isotonic 0.9% (w/v) saline for 2 minutes. Total cellular content was then extracted with 1.7mL ice-cold acetonitrile/water (50:50, v/v) solution. Cell extracts were collected using a cell scraper and quickly transferred to MagNA Lyser Green Beads tubes (Roche, Indianapolis, USA) and stored in -80°C. Media was added to empty plates and incubated together with the cells for the duration of the experiment served as a blank."},

"TREATMENT":{"TREATMENT_SUMMARY":"Monolayer cell cultures of desmoid tumors and normal fibroblasts from desmoid patients and an unaffected fibroblast cell line were grown in DMEM supplemented with 5% fetal bovine serum and maintained at 37°C in 5% CO2. Cells were divided when confluent and experiments were performed between the third and sixth passages. Approximately 10 x 106 cells were treated with 1.0uM Dasatinib (Selleck, Houston, USA) dissolved in DMSO, or 0.5uM FAK Inhibitor 14 (Cayman Chemicals Company, Michigan, USA) dissolved in water. Cells were incubated in fresh media containing the inhibitors, or vehicle, for 24 hours."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Cells were homogenized on the MagNA Lyzer, with two 30-sec cycles at 2000 rpm, resting in a -20 °C chilling block for 1 min in between pulses, and centrifuged samples at 16,000 rcf for 4 min. The cell lysate was transferred to a new 2 mL Lo-Bind Eppendorf tubes, with the final cell count approximately 10x10^6 cells for each sample. Of the twenty cell lysate samples, six samples had sufficient volume for study samples and to be included in an analytical quality control (QC) total pool. Aliquots from these cell lysate samples were combined, divided into three total pool aliquots, and processed identically to the cell lysate study samples. All study and pool samples were lyophilized to dryness and reconstituted in a 0.2M phosphate buffer, pH 7.4, in D2O with 10% Chenomx ISTD."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"-","INSTRUMENT_NAME":"-","COLUMN_NAME":"-"},

"ANALYSIS":{"ANALYSIS_TYPE":"NMR"},

"NM":{"INSTRUMENT_NAME":"Bruker Avance III","INSTRUMENT_TYPE":"FT-NMR","NMR_EXPERIMENT_TYPE":"1D-1H","SPECTROMETER_FREQUENCY":"700.13 MHz"},

"Data":[{"Bin 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