{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002208","ANALYSIS_ID":"AN003611","VERSION":"1","CREATED_ON":"July 7, 2022, 2:48 pm"},

"PROJECT":{"PROJECT_TITLE":"Metabolomic analysis to assess response to immunotherapy for malignant brain tumors: Part 2","PROJECT_TYPE":"Study of the urine and serum in mice treated with DC vaccine via 1H NMR","PROJECT_SUMMARY":"The objective of this project was to identify a peripheral metabolomic profile to serve as a biomarker of response to immunotherapy for the treatment of malignant brain tumors.","INSTITUTE":"University of Florida","DEPARTMENT":"Applied Physiology and Kinesiology","LABORATORY":"Rm 042","LAST_NAME":"Khattri","FIRST_NAME":"Ram","ADDRESS":"1864 Stadium RD, Gainesville, FL, 32611, USA","EMAIL":"rbk11@ufl.edu","PHONE":"3307856045","FUNDING_SOURCE":"This work was partially funded by the Florida Center for Brain Tumor Research (FCBTR), and by generous benefactors to the University of Florida, Peter and Angela Dziegielewski, who established the Eilzabeth Dziegielewski Glioblastoma Research Fund, and Rosalinde Wolfe, who established the Greg Wolfe Brain Tumor Research Fund. RK and MM were supported by funding from National Institutes of Health (U24-DK097209 and 5U2C-DK119889). All NMR portion of this study was performed in McKnight Brain Institute at National High Magnetic Field Laboratory’s Advanced Magnetic Resonance Imaging and Spectroscopy (AMRIS) Facility, which is funded by National Science Foundation Cooperative Agreement No. DMR-1644779 and the State of Florida.","PROJECT_COMMENTS":"Study of the urine and serum in mice treated with DC vaccine via 1H NMR","PUBLICATIONS":"Metabolomics journal (submitted)","CONTRIBUTORS":"Farhad Dastmalchi, Ram B. Khattri, Marc A. McLeod, Kaitlyn Melnick, Loic P. Deleyrolle, Yusuf Mehkri, Aida Karachi, Paul Kubilis, Shu Wang, Duane A. Mitchell, Matthew E. Merritt, Maryam Rahman"},

"STUDY":{"STUDY_TITLE":"Metabolomic analysis to assess response to immunotherapy for malignant brain tumors: Part 2","STUDY_SUMMARY":"An effective immune response in patients with cancer treated with immunotherapy includes dendritic cell (DC) activation and migration followed by stimulation of CD8 and CD4 T cells. This then leads to the activation, proliferation and further activation of other immune cell populations including NK cells or immunosuppressive populations such as Tregs and myeloid derived suppressor cells (MDSCs). These studies were carried out utilizing murine brain tumor models treated with an RNA DC vaccine platform. We hypothesized that metabolomic analyses of urines would be sensitive to the action of this diverse set of immune cells. The objective of this study was to evaluate the feasibility of using metabolomics to follow immune responses after immunotherapy. We chose NMR as our analytical technique of choice, as it has many favorable qualities that make it ideal for analyses of urine.","INSTITUTE":"University of Florida","DEPARTMENT":"Applied Physiology and Kinesiology","LABORATORY":"Rm 042","LAST_NAME":"Khattri","FIRST_NAME":"Ram","ADDRESS":"1200 Newell Dr., ARB 240, Gainesville, FL, 32611, USA","EMAIL":"rbk11@ufl.edu","NUM_GROUPS":"10","TOTAL_SUBJECTS":"80","NUM_MALES":"NA","NUM_FEMALES":"NA","STUDY_COMMENTS":"Metabolomic profiling of urine samples","PUBLICATIONS":"Metabolomics journal (submitted)","STUDY_TYPE":"Cancer surrogate biomarker discovery","PHONE":"3307856045"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"C57BL/6J","GENDER":"Not applicable","ANIMAL_ANIMAL_SUPPLIER":"Jackson Labs (Bar Harbor, ME)","ANIMAL_HOUSING":"Housed in a temperature of 22 oC","ANIMAL_LIGHT_CYCLE":"12-hour light/12-hour dark","ANIMAL_WATER":"free access to food and water (3-5 animals per cage)."},
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{
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},
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},
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},
{
"Subject ID":"-",
"Sample ID":"Maryam_2nd-set_Urine_DC_Vaccine_D9_D16",
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},
{
"Subject ID":"-",
"Sample ID":"Maryam_2nd-set_Urine_DC_Vaccine_D10_D16",
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},
{
"Subject ID":"-",
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},
{
"Subject ID":"-",
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},
{
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},
{
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},
{
"Subject ID":"-",
"Sample ID":"Maryam_2nd-set_Urine_DC_Vaccine_D5_D20",
"Factors":{"Group":"DC_Vaccine_Day 0 to 20"},
"Additional sample data":{"RAW_FILE_NAME":"Maryam_2nd-set_Urine_DC_Vaccine_D5_D20.raw"}
},
{
"Subject ID":"-",
"Sample ID":"Maryam_2nd-set_Urine_DC_Vaccine_D7_D20",
"Factors":{"Group":"DC_Vaccine_Day 0 to 20"},
"Additional sample data":{"RAW_FILE_NAME":"Maryam_2nd-set_Urine_DC_Vaccine_D7_D20.raw"}
},
{
"Subject ID":"-",
"Sample ID":"Maryam_2nd-set_Urine_DC_Vaccine_D8_D20",
"Factors":{"Group":"DC_Vaccine_Day 0 to 20"},
"Additional sample data":{"RAW_FILE_NAME":"Maryam_2nd-set_Urine_DC_Vaccine_D8_D20.raw"}
},
{
"Subject ID":"-",
"Sample ID":"Maryam_2nd-set_Urine_DC_Vaccine_D9_D20",
"Factors":{"Group":"DC_Vaccine_Day 0 to 20"},
"Additional sample data":{"RAW_FILE_NAME":"Maryam_2nd-set_Urine_DC_Vaccine_D9_D20.raw"}
},
{
"Subject ID":"-",
"Sample ID":"Maryam_2nd-set_Urine_DC_Vaccine_D10_D20",
"Factors":{"Group":"DC_Vaccine_Day 0 to 20"},
"Additional sample data":{"RAW_FILE_NAME":"Maryam_2nd-set_Urine_DC_Vaccine_D10_D20.raw"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Intakt urine samples were collected from C57/Bl6 mice. Urine is collected from a clean surface after urination.","SAMPLE_TYPE":"Urine","COLLECTION_METHOD":"Each animal was in a separate chamber individually. The chamber bottom was covered with clean parafilm. Animals were kept in the chamber until urination. Urine drops were collected using sterile syringes. Samples were stored at -80 °C until further analysis.","COLLECTION_LOCATION":"University of Florida, Neurosurgery Department, Medical college, University of Florida","COLLECTION_FREQUENCY":"Post-vaccination day 1, day 8, day 12, day 16, and day 20","COLLECTION_DURATION":"~30 minutes","VOLUMEORAMOUNT_COLLECTED":"~100 uL","STORAGE_CONDITIONS":"-80℃","COLLECTION_VIALS":"cryovials","STORAGE_VIALS":"cryovials"},

"TREATMENT":{"TREATMENT_SUMMARY":"DC vaccine treatment Naïve C57/BL6 mice were used for tumor implantation. B16F10-OVA (2 × 104 cells/brain) tumor cell lines were injected intracranially. Antigen specific T cells (3 × 107 cells/50 µL of PBS) were infused into the animals intravenous (IV) five days after tumor implantation and the DC vaccine (1x106 cells/50 µL of PBS per animal) was injected intradermally on the same day. Both antigen-specific T cells and DC vaccine were administered once. Urine samples were collected after DC vaccination at several timepoints. Finally, animals were euthanized when they reached the endpoints. Sample collection Each animal was in a separate chamber individually. The chamber bottom was covered with clean parafilm. Animals were kept in the chamber until urination. Urine drops were collected using sterile syringes. Samples were stored at -80 °C until further analysis. Serum samples were collected from peripheral blood.","ANIMAL_ANESTHESIA":"isoflurane","ANIMAL_FASTING":"non-fasted","ANIMAL_ENDP_EUTHANASIA":"Euthanasia was carried out by thoracotomy followed by cervical dislocation."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"No filtration was performed for urine samples. The urine samples were centrifuged at 4 oC, with 13.2 K rpm speed before mixing it with an internal reference and phosphate buffer system.","SAMPLEPREP_PROTOCOL_FILENAME":"DC_Vaccine_treated_NMR_urine_Procedures.docx","PROCESSING_METHOD":"None","PROCESSING_STORAGE_CONDITIONS":"-80℃","EXTRACTION_METHOD":"None","EXTRACT_STORAGE":"-80℃","SAMPLE_RESUSPENSION":"In 500 microliter of 150 mM phosphate buffer (pH 7.2) with 2 mM EDTA, 0.5 mM DSS and 0.2% sodium azide for aqueous phase samples.","SAMPLE_SPIKING":"0.5 mM DSS for urine samples"},

"ANALYSIS":{"DATA_FORMAT":"fid, 1r"},

"NM":{"INSTRUMENT_NAME":"Bruker 600 MHz","INSTRUMENT_TYPE":"FT-NMR","NMR_EXPERIMENT_TYPE":"1D-1H","FIELD_FREQUENCY_LOCK":"Deuterium","STANDARD_CONCENTRATION":"0.5mM DSS","SPECTROMETER_FREQUENCY":"600 MHz","NMR_PROBE":"Z44866_0026 (CP TXI 600S3 H-C/N-D-05 Z)","NMR_SOLVENT":"Phosphate buffer (pH 7.2) + 2 mM EDTA + 0.5 mM DSS + 0.2% of sodium azide in deuterated environment","NMR_TUBE_SIZE":"5 mm O.D.","SHIMMING_METHOD":"Topshim","PULSE_SEQUENCE":"noesypr1d","WATER_SUPPRESSION":"presat","PULSE_WIDTH":"90-degree","RECEIVER_GAIN":"101","OFFSET_FREQUENCY":"4.78 ppm","CHEMICAL_SHIFT_REF_CPD":"DSS","TEMPERATURE":"298.2 oK","NUMBER_OF_SCANS":"64","DUMMY_SCANS":"8","ACQUISITION_TIME":"4s","RELAXATION_DELAY":"1s","SPECTRAL_WIDTH":"7142.9 Hz","NUM_DATA_POINTS_ACQUIRED":"28572","REAL_DATA_POINTS":"65536","LINE_BROADENING":"0.22 Hz","ZERO_FILLING":"65,536 points","APODIZATION":"Exponential","BASELINE_CORRECTION_METHOD":"Spline","CHEMICAL_SHIFT_REF_STD":"0 ppm for DSS"},

"NMR_METABOLITE_DATA":{
"Units":"mM",

"Data":[{"Metabolite":"Formate","Maryam_2nd-set_Urine_Control_C1_Post":"0.0050","Maryam_2nd-set_Urine_Control_C2_Post":"0.0077","Maryam_2nd-set_Urine_Control_C3_Post":"0.0157","Maryam_2nd-set_Urine_Control_C4_Post":"0.0263","Maryam_2nd-set_Urine_Control_C5_Post":"0.0379","Maryam_2nd-set_Urine_Control_C6_Post":"0.0339","Maryam_2nd-set_Urine_Control_C7_Post":"0.0382","Maryam_2nd-set_Urine_Control_C1_D8":"0.0030","Maryam_2nd-set_Urine_Control_C2_D8":"0.0079","Maryam_2nd-set_Urine_Control_C3_D8":"0.0189","Maryam_2nd-set_Urine_Control_C4_D8":"0.0202","Maryam_2nd-set_Urine_Control_C5_D8":"0.0289","Maryam_2nd-set_Urine_Control_C6_D8":"0.0214","Maryam_2nd-set_Urine_Control_C7_D8":"0.0102","Maryam_2nd-set_Urine_Control_C1_D12":"0.0100","Maryam_2nd-set_Urine_Control_C2_D12":"0.0100","Maryam_2nd-set_Urine_Control_C3_D12":"0.0287","Maryam_2nd-set_Urine_Control_C4_D12":"0.0128","Maryam_2nd-set_Urine_Control_C5_D12":"0.0152","Maryam_2nd-set_Urine_Control_C6_D12":"0.0193","Maryam_2nd-set_Urine_Control_C7_D12":"0.0095","Maryam_2nd-set_Urine_Control_C1_D16":"0.0126","Maryam_2nd-set_Urine_Control_C2_D16":"0.0128","Maryam_2nd-set_Urine_Control_C3_D16":"0.0083","Maryam_2nd-set_Urine_Control_C4_D16":"0.0361","Maryam_2nd-set_Urine_Control_C5_D16":"0.0145","Maryam_2nd-set_Urine_Control_C7_D16":"0.0069","Maryam_2nd-set_Urine_Control_C1_D20":"0.0099","Maryam_2nd-set_Urine_Control_C2_D20":"0.0081","Maryam_2nd-set_Urine_Control_C3_D20":"0.0108","Maryam_2nd-set_Urine_Control_C7_D20":"0.0046","Maryam_2nd-set_Urine_DC_Vaccine_D1_Post":"0.0338","Maryam_2nd-set_Urine_DC_Vaccine_D2_Post":"0.0209","Maryam_2nd-set_Urine_DC_Vaccine_D3_Post":"0.0188","Maryam_2nd-set_Urine_DC_Vaccine_D4_Post":"0.0192","Maryam_2nd-set_Urine_DC_Vaccine_D5_Post":"0.0179","Maryam_2nd-set_Urine_DC_Vaccine_D6_Post":"0.0258","Maryam_2nd-set_Urine_DC_Vaccine_D7_Post":"0.0213","Maryam_2nd-set_Urine_DC_Vaccine_D8_Post":"0.0102","Maryam_2nd-set_Urine_DC_Vaccine_D9_Post":"0.0121","Maryam_2nd-set_Urine_DC_Vaccine_D10_Post":"0.0328","Maryam_2nd-set_Urine_DC_Vaccine_D1_D8":"0.0740","Maryam_2nd-set_Urine_DC_Vaccine_D2_D8":"0.0215","Maryam_2nd-set_Urine_DC_Vaccine_D3_D8":"0.0466","Maryam_2nd-set_Urine_DC_Vaccine_D4_D8":"0.0641","Maryam_2nd-set_Urine_DC_Vaccine_D5_D8":"0.0204","Maryam_2nd-set_Urine_DC_Vaccine_D6_D8":"0.0099","Maryam_2nd-set_Urine_DC_Vaccine_D7_D8":"0.0360","Maryam_2nd-set_Urine_DC_Vaccine_D8_D8":"0.0168","Maryam_2nd-set_Urine_DC_Vaccine_D9_D8":"0.0467","Maryam_2nd-set_Urine_DC_Vaccine_D10_D8":"0.0315","Maryam_2nd-set_Urine_DC_Vaccine_D1_D12":"0.0131","Maryam_2nd-set_Urine_DC_Vaccine_D2_D12":"0.0212","Maryam_2nd-set_Urine_DC_Vaccine_D3_D12":"0.0161","Maryam_2nd-set_Urine_DC_Vaccine_D4_D12":"0.0081","Maryam_2nd-set_Urine_DC_Vaccine_D5_D12":"0.0111","Maryam_2nd-set_Urine_DC_Vaccine_D6_D12":"0.0066","Maryam_2nd-set_Urine_DC_Vaccine_D7_D12":"0.0180","Maryam_2nd-set_Urine_DC_Vaccine_D8_D12":"0.0028","Maryam_2nd-set_Urine_DC_Vaccine_D9_D12":"0.0080","Maryam_2nd-set_Urine_DC_Vaccine_D10_D12":"0.0139","Maryam_2nd-set_Urine_DC_Vaccine_D1_D16":"0.0062","Maryam_2nd-set_Urine_DC_Vaccine_D2_D16":"0.0037","Maryam_2nd-set_Urine_DC_Vaccine_D3_D16":"0.0074","Maryam_2nd-set_Urine_DC_Vaccine_D4_D16":"0.0049","Maryam_2nd-set_Urine_DC_Vaccine_D5_D16":"0.0078","Maryam_2nd-set_Urine_DC_Vaccine_D6_D16":"0.0348","Maryam_2nd-set_Urine_DC_Vaccine_D7_D16":"0.0026","Maryam_2nd-set_Urine_DC_Vaccine_D8_D16":"0.0162","Maryam_2nd-set_Urine_DC_Vaccine_D9_D16":"0.0165","Maryam_2nd-set_Urine_DC_Vaccine_D10_D16":"0.0202","Maryam_2nd-set_Urine_DC_Vaccine_D1_D20":"0.0149","Maryam_2nd-set_Urine_DC_Vaccine_D2_D20":"0.0068","Maryam_2nd-set_Urine_DC_Vaccine_D3_D20":"0.0118","Maryam_2nd-set_Urine_DC_Vaccine_D4_D20":"0.0020","Maryam_2nd-set_Urine_DC_Vaccine_D5_D20":"0.0127","Maryam_2nd-set_Urine_DC_Vaccine_D7_D20":"0.0228","Maryam_2nd-set_Urine_DC_Vaccine_D8_D20":"0.0076","Maryam_2nd-set_Urine_DC_Vaccine_D9_D20":"0.0108","Maryam_2nd-set_Urine_DC_Vaccine_D10_D20":"0.0143"},{"Metabolite":"3-Indoxylsulfate","Maryam_2nd-set_Urine_Control_C1_Post":"0.0448","Maryam_2nd-set_Urine_Control_C2_Post":"0.0406","Maryam_2nd-set_Urine_Control_C3_Post":"0.2322","Maryam_2nd-set_Urine_Control_C4_Post":"0.2005","Maryam_2nd-set_Urine_Control_C5_Post":"0.1829","Maryam_2nd-set_Urine_Control_C6_Post":"0.2959","Maryam_2nd-set_Urine_Control_C7_Post":"0.1768","Maryam_2nd-set_Urine_Control_C1_D8":"0.1256","Maryam_2nd-set_Urine_Control_C2_D8":"0.1125","Maryam_2nd-set_Urine_Control_C3_D8":"0.1906","Maryam_2nd-set_Urine_Control_C4_D8":"0.0375","Maryam_2nd-set_Urine_Control_C5_D8":"0.1301","Maryam_2nd-set_Urine_Control_C6_D8":"0.2063","Maryam_2nd-set_Urine_Control_C7_D8":"0.1120","Maryam_2nd-set_Urine_Control_C1_D12":"0.1895","Maryam_2nd-set_Urine_Control_C2_D12":"0.1252","Maryam_2nd-set_Urine_Control_C3_D12":"0.0745","Maryam_2nd-set_Urine_Control_C4_D12":"0.1378","Maryam_2nd-set_Urine_Control_C5_D12":"0.2378","Maryam_2nd-set_Urine_Control_C6_D12":"0.1676","Maryam_2nd-set_Urine_Control_C7_D12":"0.2601","Maryam_2nd-set_Urine_Control_C1_D1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"Metabolites":[{"metabolite_name":"Formate"},{"metabolite_name":"3-Indoxylsulfate"},{"metabolite_name":"Hippurate"},{"metabolite_name":"Phenylalanine"},{"metabolite_name":"Histidine"},{"metabolite_name":"Tyrosine"},{"metabolite_name":"3-Hydroxyphenylacetate"},{"metabolite_name":"Cis-acotinate"},{"metabolite_name":"allantoin"},{"metabolite_name":"Glucose"},{"metabolite_name":"N-Acetylglucosamine"},{"metabolite_name":"Taurine"},{"metabolite_name":"Choline"},{"metabolite_name":"Dimethyl sulfone"},{"metabolite_name":"Creatinine"},{"metabolite_name":"Creatine"},{"metabolite_name":"N,N-Dimethyl glycine"},{"metabolite_name":"Citrate"},{"metabolite_name":"2-Oxoglutarate"},{"metabolite_name":"Succinate"},{"metabolite_name":"N-Carbamoyl-ß-Alanine"},{"metabolite_name":"Acetate"},{"metabolite_name":"Alanine"},{"metabolite_name":"Lactate"},{"metabolite_name":"3-Hydroxybutyrate"},{"metabolite_name":"Propylene glycol"},{"metabolite_name":"Isobutyrate"},{"metabolite_name":"3-Hydroxyisobutyrate"},{"metabolite_name":"Isoleucine"},{"metabolite_name":"Leucine"},{"metabolite_name":"2-oxo isocaproate"},{"metabolite_name":"2-Hydroxybutyrate"},{"metabolite_name":"2-Hydroxy-3-methylvalerate"}]
}

}