#METABOLOMICS WORKBENCH japizsaab_20230324_031900 DATATRACK_ID:3815 STUDY_ID:ST002529 ANALYSIS_ID:AN004163 PROJECT_ID:PR001627 VERSION 1 CREATED_ON March 28, 2023, 10:11 am #PROJECT PR:PROJECT_TITLE Pancreatic tumors activate arginine biosynthesis to adapt to myeloid-driven PR:PROJECT_TITLE amino acid stress PR:PROJECT_SUMMARY Nutrient stress in the tumor microenvironment requires cancer cells to adopt PR:PROJECT_SUMMARY adaptive metabolic programs to maintain survival and proliferation. Therefore, PR:PROJECT_SUMMARY knowledge of microenvironmental nutrient levels and how cancer cells cope with PR:PROJECT_SUMMARY such nutrition is critical to understand the metabolism underpinning cancer cell PR:PROJECT_SUMMARY biology. Previously, we performed quantitative metabolomics of the interstitial PR:PROJECT_SUMMARY fluid (the local perfusate) of murine pancreatic ductal adenocarcinoma (PDAC) PR:PROJECT_SUMMARY tumors to comprehensively characterize nutrient availability in the PR:PROJECT_SUMMARY microenvironment of these tumors (Sullivan et al., 2019a). Here, we develop PR:PROJECT_SUMMARY Tumor Interstitial Fluid Medium (TIFM), a cell culture medium that contains PR:PROJECT_SUMMARY nutrient levels representative of the PDAC microenvironment, enabling study of PR:PROJECT_SUMMARY PDAC metabolism under physiological nutrition. We show that PDAC cells cultured PR:PROJECT_SUMMARY in TIFM, compared to standard laboratory models, adopt a cellular state more PR:PROJECT_SUMMARY similar to PDAC cells in tumors. Further, using the TIFM model we identified PR:PROJECT_SUMMARY arginine biosynthesis as a metabolic adaptation PDAC cells engage to cope with PR:PROJECT_SUMMARY microenvironmental arginine starvation driven by myeloid cells in PDAC tumors. PR:PROJECT_SUMMARY Altogether, these data show that nutrient availability in tumors is an important PR:PROJECT_SUMMARY determinant of cancer cell metabolism and behavior, and cell culture models that PR:PROJECT_SUMMARY incorporate physiological nutrient availability have improved fidelity and PR:PROJECT_SUMMARY enable the discovery of novel cancer metabolic phenotypes. PR:INSTITUTE University of Chicago PR:LAST_NAME Apiz Saab PR:FIRST_NAME Juan PR:ADDRESS 929 E. 57th St. PR:EMAIL japizsaab@uchicago.edu PR:PHONE 7738346506 #STUDY ST:STUDY_TITLE In vivo 15N2-glutamine tracing by jugular vein infusion in PDAC-tumor bearing ST:STUDY_TITLE Lyz2-Arg1 and control mice ST:STUDY_SUMMARY We assessed the relative levels of urea cycle related metabolites in PDAC tumors ST:STUDY_SUMMARY in Lyz2-Cre+/+; Arg1fl/fl compared to Arg1fl/fl control host animals. ST:INSTITUTE University of Chicago ST:LAST_NAME Apiz Saab ST:FIRST_NAME Juan ST:ADDRESS 929 E. 57th St. ST:EMAIL japizsaab@uchicago.edu ST:PHONE 7738346506 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - 0785-0 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0785-0.mzML SUBJECT_SAMPLE_FACTORS - 0785-120 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0785-120.mzML SUBJECT_SAMPLE_FACTORS - 0785-240 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0785-240.mzML SUBJECT_SAMPLE_FACTORS - 0785-30 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0785-30.mzML SUBJECT_SAMPLE_FACTORS - 0787-0 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0787-0.mzML SUBJECT_SAMPLE_FACTORS - 0787-120 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0787-120.mzML SUBJECT_SAMPLE_FACTORS - 0787-240 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0787-240.mzML SUBJECT_SAMPLE_FACTORS - 0787-30 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0787-30.mzML SUBJECT_SAMPLE_FACTORS - 0788-0 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0788-0.mzML SUBJECT_SAMPLE_FACTORS - 0788-120 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0788-120.mzML SUBJECT_SAMPLE_FACTORS - 0788-240 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0788-240.mzML SUBJECT_SAMPLE_FACTORS - 0788-30 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0788-30.mzML SUBJECT_SAMPLE_FACTORS - 0789-0 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0789-0.mzML SUBJECT_SAMPLE_FACTORS - 0789-120 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0789-120.mzML SUBJECT_SAMPLE_FACTORS - 0789-240 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0789-240.mzML SUBJECT_SAMPLE_FACTORS - 0789-30 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_0789-30.mzML SUBJECT_SAMPLE_FACTORS - A35066-0 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35066-0.mzML SUBJECT_SAMPLE_FACTORS - A35066-120 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35066-120.mzML SUBJECT_SAMPLE_FACTORS - A35066-240 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35066-240.mzML SUBJECT_SAMPLE_FACTORS - A35066-30 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35066-30.mzML SUBJECT_SAMPLE_FACTORS - A35089-0 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35089-0.mzML SUBJECT_SAMPLE_FACTORS - A35089-120 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35089-120.mzML SUBJECT_SAMPLE_FACTORS - A35089-240 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35089-240.mzML SUBJECT_SAMPLE_FACTORS - A35089-30 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35089-30.mzML SUBJECT_SAMPLE_FACTORS - A35096-0 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35096-0.mzML SUBJECT_SAMPLE_FACTORS - A35096-120 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35096-120.mzML SUBJECT_SAMPLE_FACTORS - A35096-240 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35096-240.mzML SUBJECT_SAMPLE_FACTORS - A35096-30 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35096-30.mzML SUBJECT_SAMPLE_FACTORS - A35097-0 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35097-0.mzML SUBJECT_SAMPLE_FACTORS - A35097-120 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35097-120.mzML SUBJECT_SAMPLE_FACTORS - A35097-240 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35097-240.mzML SUBJECT_SAMPLE_FACTORS - A35097-30 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35097-30.mzML SUBJECT_SAMPLE_FACTORS - A35098-0 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35098-0.mzML SUBJECT_SAMPLE_FACTORS - A35098-120 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35098-120.mzML SUBJECT_SAMPLE_FACTORS - A35098-240 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35098-240.mzML SUBJECT_SAMPLE_FACTORS - A35098-30 Sample type:Plasma | Condition:Arg1fl/fl RAW_FILE_NAME=Plasma_A35098-30.mzML SUBJECT_SAMPLE_FACTORS - A35465-0 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35465-0.mzML SUBJECT_SAMPLE_FACTORS - A35465-120 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35465-120.mzML SUBJECT_SAMPLE_FACTORS - A35465-240 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35465-240.mzML SUBJECT_SAMPLE_FACTORS - A35465-30 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35465-30.mzML SUBJECT_SAMPLE_FACTORS - A35469-0 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35469-0.mzML SUBJECT_SAMPLE_FACTORS - A35469-120 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35469-120.mzML SUBJECT_SAMPLE_FACTORS - A35469-240 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35469-240.mzML SUBJECT_SAMPLE_FACTORS - A35469-30 Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=Plasma_A35469-30.mzML SUBJECT_SAMPLE_FACTORS - 0785_tumor_Arg1_KO Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=0785_tumor_Arg1_KO.mzML SUBJECT_SAMPLE_FACTORS - 0787_tumor_Arg1_KO Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=0787_tumor_Arg1_KO.mzML SUBJECT_SAMPLE_FACTORS - 0788_tumor_Arg1_KO Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=0788_tumor_Arg1_KO.mzML SUBJECT_SAMPLE_FACTORS - 0789_tumor_Arg1_KO Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=0789_tumor_Arg1_KO.mzML SUBJECT_SAMPLE_FACTORS - A35089_tumor_Arg1_KO Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=A35089_tumor_Arg1_KO.mzML SUBJECT_SAMPLE_FACTORS - A35096_tumor_Control Sample type:Tumor | Condition:Arg1fl/fl RAW_FILE_NAME=A35096_tumor_Control.mzML SUBJECT_SAMPLE_FACTORS - A35097_tumor_Control Sample type:Tumor | Condition:Arg1fl/fl RAW_FILE_NAME=A35097_tumor_Control.mzML SUBJECT_SAMPLE_FACTORS - A35098_tumor_Control Sample type:Tumor | Condition:Arg1fl/fl RAW_FILE_NAME=A35098_tumor_Control.mzML SUBJECT_SAMPLE_FACTORS - A35465_tumor_Arg1_KO Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=A35465_tumor_Arg1_KO.mzML SUBJECT_SAMPLE_FACTORS - A35466_tumor_Arg1_KO Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=A35466_tumor_Arg1_KO.mzML SUBJECT_SAMPLE_FACTORS - A35469_tumor_Arg1_KO Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl RAW_FILE_NAME=A35469_tumor_Arg1_KO.mzML #COLLECTION CO:COLLECTION_SUMMARY 6~8-month-old female & male Lyz2-Cre +/+; Arg1fl/fl and litter mate control CO:COLLECTION_SUMMARY Arg1fl/fl mice with murine PDAC orthotopic tumors underwent dual jugular vein & CO:COLLECTION_SUMMARY carotid artery catheterization surgery. On day 5 of post recovery, mice received CO:COLLECTION_SUMMARY a 0.28 mg/g 10 min. bolus followed by a continuous 4 hr. infusion 0.005 mg/g/min CO:COLLECTION_SUMMARY infusion of 15N2-glutamine (Cambridge Isotope Laboratory #NLM-1328-PK). Plasma CO:COLLECTION_SUMMARY samples were taken at time points: 0' 15' 30' 60' 120' 180’ 240’. Tumors and CO:COLLECTION_SUMMARY tissues were harvested at 240’ and immediately snap frozen with liquid CO:COLLECTION_SUMMARY nitrogen stored at -80°C prior to analysis. CO:SAMPLE_TYPE Tumor, plasma #TREATMENT TR:TREATMENT_SUMMARY 6~8-month-old female & male Lyz2-Cre +/+; Arg1fl/fl and litter mate control TR:TREATMENT_SUMMARY Arg1fl/fl mice with murine PDAC orthotopic tumors underwent dual jugular vein & TR:TREATMENT_SUMMARY carotid artery catheterization surgery. On day 5 of post recovery, mice received TR:TREATMENT_SUMMARY a 0.28 mg/g 10 min. bolus followed by a continuous 4 hr. infusion 0.005 mg/g/min TR:TREATMENT_SUMMARY infusion of 15N2-glutamine (Cambridge Isotope Laboratory #NLM-1328-PK). Plasma TR:TREATMENT_SUMMARY samples were taken at time points: 0' 15' 30' 60' 120' 180’ 240’. Tumors and TR:TREATMENT_SUMMARY tissues were harvested at 240’ and immediately snap frozen with liquid TR:TREATMENT_SUMMARY nitrogen stored at -80°C prior to analysis. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Cryogenically frozen tumor pieces were ground to a fine homogenous powder with a SP:SAMPLEPREP_SUMMARY liquid nitrogen cooled mortar and pestle. ~30mg of tissue powder was weighed SP:SAMPLEPREP_SUMMARY into sample tubes, and metabolites were extracted with 600µL HPLC grade SP:SAMPLEPREP_SUMMARY methanol, 300µL HPLC grade water, and 400µL chloroform. Samples were vortexed SP:SAMPLEPREP_SUMMARY for 10min at 4°C, centrifuged 21,000xg at 4°C for 10 min. 400µL of the SP:SAMPLEPREP_SUMMARY aqueous top layer was removed into a new tube and dried under nitrogen. Dried SP:SAMPLEPREP_SUMMARY tumor extracts were resuspended in 100µL HPLC grade water and LC-MS analysis SP:SAMPLEPREP_SUMMARY was performed as described before(Sullivan et al., 2019b, 2019a). For plasma SP:SAMPLEPREP_SUMMARY samples, we extracted polar metabolites from 5µL of sample using 45µL of a SP:SAMPLEPREP_SUMMARY 75:25:0.1 HPLC grade acetonitrile:methanol:formic acid extraction mix. Samples SP:SAMPLEPREP_SUMMARY in extraction mix were vortexed for 10 min at 4°C and centrifugated at 15,000x SP:SAMPLEPREP_SUMMARY rpm for 10 min at 4°C to pellet insoluble material. 20µL of the soluble polar SP:SAMPLEPREP_SUMMARY metabolite supernatant was moved to sample vials for analysis by LC-MS. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Atlantis BEH Z-HILIC (2.1x150 mm, 2.5 µM; part # 186009990; Waters Corporation) CH:SOLVENT_A 20 mM ammonium bicarbonate at pH 9.6, adjusted by ammonium hydroxide addition CH:SOLVENT_B 100% Acetonitrile CH:FLOW_GRADIENT 0 minute: 85% B, 0.5 minute: 85% B, 18 minutes: 20% B, 20 minutes: 20% B, 20.5 CH:FLOW_GRADIENT minutes: 85% B and 28 minutes: 85% B CH:FLOW_RATE 0.2 mL/minute CH:COLUMN_TEMPERATURE 30°C #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap IQ-X Tribrid MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS Data acquisition was done using the Xcalibur software (Thermo Scientific) in MS:MS_COMMENTS full-scan mode with a range of 70-1000 m/z at 60K. Metabolite identification was MS:MS_COMMENTS done by matching the retention time and MS/MS fragmentation to the reference MS:MS_COMMENTS standards. Data analysis was performed using Tracefinder 5.1 software (Thermo MS:MS_COMMENTS Scientific). #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Natural abundance corrected percentages MS_METABOLITE_DATA_START Samples A35096_tumor_Control A35097_tumor_Control A35098_tumor_Control 0785_tumor_Arg1_KO 0787_tumor_Arg1_KO 0788_tumor_Arg1_KO 0789_tumor_Arg1_KO A35089_tumor_Arg1_KO A35465_tumor_Arg1_KO A35466_tumor_Arg1_KO A35469_tumor_Arg1_KO A35096-0 A35096-120 A35096-240 A35096-30 A35097-0 A35097-120 A35097-240 A35097-30 A35098-0 A35098-120 A35098-240 A35098-30 0785-0 0785-120 0785-240 0785-30 0787-0 0787-120 0787-240 0787-30 0788-0 0788-120 0788-240 0788-30 0789-0 0789-120 0789-240 0789-30 A35066-0 A35066-120 A35066-240 A35066-30 A35089-0 A35089-120 A35089-240 A35089-30 A35465-0 A35465-120 A35465-240 A35465-30 A35469-0 A35469-120 A35469-240 A35469-30 Factors Sample type:Tumor | Condition:Arg1fl/fl Sample type:Tumor | Condition:Arg1fl/fl Sample type:Tumor | Condition:Arg1fl/fl Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Tumor | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Sample type:Plasma | Condition:Lyz2-Cre+/+; Arg1fl/fl Glutamine_M+0 42.03 45.24 41.12 36.60 30.69 36.96 48.20 37.68 40.51 35.02 33.44 99.95 38.14 35.86 42.61 99.95 37.27 37.04 38.67 100.00 32.13 32.55 32.60 99.95 33.04 25.56 38.99 99.96 23.55 20.70 32.12 99.86 31.03 21.26 34.21 100.00 41.92 6.84 43.51 100.00 31.22 23.15 34.62 99.97 34.52 29.49 36.02 100.00 40.35 32.26 40.82 100.00 26.40 22.23 30.05 Glutamine_M+1 25.06 25.27 24.09 27.70 22.83 22.14 26.53 29.98 25.68 26.41 23.28 0.00 26.45 26.79 20.22 0.00 25.23 26.66 19.59 0.00 23.01 24.83 21.98 0.00 25.10 27.99 21.64 0.00 18.71 22.46 13.50 0.00 22.37 22.27 22.71 0.00 24.73 1.41 22.23 0.00 24.61 25.13 19.64 0.00 27.87 29.77 25.82 0.00 23.00 25.96 22.18 0.00 20.85 24.23 19.41 Glutamine_M+2 32.91 29.49 34.80 35.71 46.48 40.91 25.27 32.34 33.81 38.57 43.28 0.05 35.41 37.35 37.17 0.05 37.50 36.31 41.74 0.00 44.86 42.62 45.42 0.05 41.85 46.46 39.37 0.04 57.74 56.84 54.38 0.14 46.59 56.47 43.08 0.00 33.35 91.75 34.26 0.00 44.17 51.72 45.75 0.03 37.61 40.74 38.17 0.00 36.65 41.78 37.00 0.00 52.76 53.54 50.54 Aspartate_M+0 76.75 78.29 78.06 82.19 75.45 83.00 81.34 77.43 78.31 76.87 77.81 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 99.98 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 Aspartate_M+1 23.25 21.71 21.94 17.81 24.55 17.00 18.66 22.57 21.69 23.13 22.19 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Citrulline _M+0 64.07 62.47 60.93 65.11 43.44 74.62 58.31 66.67 58.17 63.77 60.02 100.00 43.93 43.12 58.91 100.00 43.37 42.37 52.31 100.00 39.33 40.36 42.16 100.00 38.38 29.34 48.21 100.00 29.59 26.12 44.28 100.00 44.56 34.06 51.03 100.00 51.60 43.52 52.86 100.00 36.95 27.23 48.39 100.00 43.31 34.42 49.91 100.00 49.12 36.50 49.87 100.00 37.78 29.09 47.06 Citrulline _M+1 26.30 26.94 28.42 22.76 32.33 16.79 31.23 22.66 28.33 21.91 23.40 0.00 38.45 39.95 31.58 0.00 39.62 40.58 33.18 0.00 40.82 42.26 36.84 0.00 41.89 41.42 37.03 0.00 39.41 40.72 32.60 0.00 38.17 39.74 33.82 0.00 37.56 40.94 34.08 0.00 40.03 39.42 35.04 0.00 39.53 41.47 34.02 0.00 37.76 40.78 34.05 0.00 38.03 39.16 33.64 Citrulline _M+2 8.41 9.16 9.06 10.07 19.14 6.96 9.39 8.86 11.41 11.58 13.02 0.00 15.15 14.48 8.27 0.00 14.49 14.63 12.18 0.00 16.61 14.70 16.74 0.00 16.78 23.46 12.71 0.00 23.94 25.32 17.62 0.00 14.43 20.14 12.41 0.00 9.64 13.52 11.34 0.00 19.17 26.07 13.82 0.00 14.60 19.81 13.32 0.00 11.62 18.97 13.60 0.00 18.99 23.89 15.23 Citrulline _M+3 1.21 1.43 1.59 2.05 5.08 1.64 1.07 1.80 2.08 2.73 3.56 0.00 2.47 2.45 1.24 0.00 2.51 2.43 2.32 0.00 3.24 2.67 4.26 0.00 2.95 5.78 2.05 0.00 7.06 7.84 5.50 0.00 2.85 6.06 2.75 0.00 1.20 2.02 1.72 0.00 3.85 7.28 2.76 0.00 2.55 4.30 2.75 0.00 1.50 3.75 2.49 0.00 5.20 7.87 4.07 Glutamate _M+0 76.73 79.22 78.34 79.06 76.37 83.23 81.73 77.39 77.82 76.94 78.24 100.00 81.63 79.96 86.25 100.00 82.74 78.48 85.98 100.00 80.65 78.00 83.88 100.00 78.71 74.21 85.47 99.60 76.40 71.23 81.67 100.00 74.90 64.24 79.08 100.00 82.85 100.00 85.08 100.00 80.10 70.62 83.28 100.00 77.62 72.28 80.50 100.00 83.32 77.54 85.12 99.97 79.19 70.61 82.03 Glutamate _M+1 23.27 20.78 21.66 20.94 23.63 16.77 18.27 22.61 22.18 23.06 21.76 0.00 18.37 20.04 13.75 0.00 17.26 21.52 14.02 0.00 19.35 22.00 16.12 0.00 21.29 25.79 14.53 0.40 23.60 28.77 18.33 0.00 25.10 35.76 20.92 0.00 17.15 0.00 14.92 0.00 19.90 29.38 16.72 0.00 22.38 27.72 19.50 0.00 16.68 22.46 14.88 0.03 20.81 29.39 17.97 Argininosuccinate _M+0 72.66 70.79 73.83 76.01 56.96 77.12 81.99 85.38 74.60 76.66 82.65 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Argininosuccinate _M+1 22.14 19.13 18.35 16.84 21.54 16.78 10.73 6.57 17.87 12.59 8.51 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Argininosuccinate _M+2 4.36 8.96 7.24 5.37 16.99 5.42 6.86 6.91 6.74 9.15 5.36 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Argininosuccinate _M+3 0.84 1.12 0.58 1.78 4.50 0.68 0.42 1.14 0.79 1.60 3.48 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Argininosuccinate _M+4 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Arginine _M+0 87.14 88.08 89.23 82.77 81.19 85.69 86.42 87.84 82.57 87.16 87.79 100.00 81.01 80.07 88.45 100.00 81.86 80.19 86.64 100.00 81.43 79.91 82.85 100.00 76.91 71.38 83.16 100.00 76.16 68.76 90.48 100.00 77.19 68.05 84.21 100.00 81.59 78.50 83.86 100.00 81.44 76.60 86.47 100.00 84.63 80.95 87.23 100.00 81.70 75.21 84.40 100.00 81.17 73.79 84.14 Arginine _M+1 8.65 7.67 6.96 9.63 8.96 8.95 8.98 7.35 10.60 6.99 6.45 0.00 12.22 12.96 8.49 0.00 11.70 12.77 8.71 0.00 11.54 12.99 10.06 0.00 13.82 14.93 10.96 0.00 11.90 14.84 5.26 0.00 15.31 18.56 10.57 0.00 12.69 14.04 10.72 0.00 10.81 11.72 8.37 0.00 9.55 10.97 7.82 0.00 12.52 14.51 9.56 0.00 10.49 13.26 9.55 Arginine _M+2 3.39 3.42 3.04 5.57 6.73 4.13 3.73 3.68 5.26 4.16 4.00 0.00 5.36 5.58 2.57 0.00 5.19 5.64 3.63 0.00 5.48 5.70 5.36 0.00 7.21 9.97 4.77 0.00 8.19 11.16 2.87 0.00 6.20 10.38 4.25 0.00 4.74 6.04 4.33 0.00 5.87 8.23 3.90 0.00 4.61 6.14 3.72 0.00 4.75 7.79 4.61 0.00 6.12 9.08 4.67 Arginine _M+3 0.73 0.74 0.67 1.77 2.62 1.07 0.79 1.01 1.41 1.46 1.49 0.00 1.26 1.23 0.45 0.00 1.13 1.26 0.90 0.00 1.39 1.27 1.53 0.00 1.85 3.19 0.99 0.00 3.12 4.34 1.14 0.00 1.16 2.59 0.84 0.00 0.90 1.29 0.98 0.00 1.65 2.92 1.10 0.00 1.06 1.67 1.06 0.00 0.95 2.19 1.26 0.00 1.88 3.22 1.42 Ornithine _M+0 93.09 94.67 93.99 91.01 86.64 92.65 92.29 92.16 89.30 90.69 92.40 100.00 87.76 87.94 91.15 100.00 89.63 88.51 90.92 100.00 88.82 88.60 88.02 100.00 85.77 77.53 86.64 100.00 76.21 72.03 89.46 100.00 85.69 79.98 87.87 100.00 88.45 86.20 86.27 100.00 82.39 76.96 88.88 100.00 86.40 82.57 89.35 100.00 85.60 79.05 84.87 100.00 85.76 79.96 89.67 Ornithine _M+1 5.73 4.37 4.92 7.13 9.83 5.46 6.65 6.28 8.82 7.12 5.58 0.00 10.02 9.95 7.67 0.00 8.39 9.02 7.23 0.00 8.94 9.15 8.92 0.00 11.82 17.39 11.40 0.00 17.00 20.44 8.03 0.00 11.16 14.38 9.28 0.00 10.13 11.35 11.87 0.00 13.78 17.32 8.85 0.00 11.36 13.62 8.53 0.00 12.46 16.87 12.67 0.00 10.88 14.52 7.76 Ornithine _M+2 1.18 0.96 1.08 1.86 3.53 1.89 1.06 1.57 1.88 2.19 2.02 0.00 2.23 2.10 1.18 0.00 1.98 2.47 1.85 0.00 2.24 2.25 3.06 0.00 2.41 5.08 1.95 0.00 6.79 7.53 2.51 0.00 3.15 5.64 2.85 0.00 1.43 2.45 1.86 0.00 3.82 5.71 2.26 0.00 2.24 3.81 2.11 0.00 1.94 4.08 2.46 0.00 3.36 5.52 2.57 Carbamoyl_aspartate _M+0 58.43 56.22 47.74 39.83 47.48 51.46 53.28 54.78 47.82 49.21 62.62 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Carbamoyl_aspartate _M+1 35.10 36.91 43.34 47.14 42.04 39.50 40.68 38.17 43.36 41.72 32.11 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Carbamoyl_aspartate _M+2 6.46 6.87 8.92 13.04 10.48 9.03 6.04 7.05 8.81 9.06 5.27 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Dihydroorotate _M+0 56.56 53.47 47.02 37.55 44.59 49.51 52.44 53.01 45.98 47.13 61.06 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Dihydroorotate _M+1 36.85 39.37 44.19 49.85 44.36 41.83 41.23 39.75 45.18 43.17 33.52 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Dihydroorotate _M+2 6.60 7.16 8.79 12.61 11.05 8.66 6.32 7.24 8.84 9.70 5.42 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Orotate _M+0 55.24 59.60 50.32 44.80 47.12 64.80 56.71 53.39 49.08 50.46 58.13 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Orotate _M+1 37.61 34.20 41.64 45.21 42.55 30.03 37.35 38.62 42.64 40.88 35.45 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Orotate _M+2 7.16 6.19 8.04 9.99 10.33 5.17 5.94 7.99 8.28 8.66 6.43 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA UMP _M+0 98.14 97.15 95.34 100.00 96.45 100.00 99.40 99.25 97.78 93.57 100.00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA UMP _M+1 1.00 1.92 3.36 0.00 2.40 0.00 0.35 0.00 1.62 5.08 0.00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA UMP _M+2 0.86 0.93 1.30 0.00 1.15 0.00 0.24 0.75 0.60 1.35 0.00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA UDP _M+0 91.60 93.41 92.06 97.79 95.66 100.00 93.31 98.49 91.82 95.17 100.00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA UDP _M+1 8.40 6.59 7.94 2.21 4.34 0.00 6.69 1.51 8.18 4.83 0.00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA UDP _M+2 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA UTP _M+0 90.48 92.12 90.02 93.80 95.18 100.00 91.54 100.00 91.41 100.00 100.00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA UTP _M+1 9.52 7.88 9.98 6.20 4.82 0.00 8.46 0.00 8.59 0.00 0.00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA UTP _M+2 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Putrescine _M+0 98.02 97.47 98.13 96.19 97.53 99.39 96.74 98.45 96.00 96.61 98.58 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Putrescine _M+1 1.89 2.30 1.84 3.30 2.09 0.59 3.04 1.34 3.73 2.88 1.06 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Putrescine _M+2 0.09 0.23 0.04 0.51 0.39 0.02 0.22 0.20 0.27 0.51 0.37 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Glutamine_M+0 Glutamine_M+1 Glutamine_M+2 Aspartate_M+0 Aspartate_M+1 Citrulline _M+0 Citrulline _M+1 Citrulline _M+2 Citrulline _M+3 Glutamate _M+0 Glutamate _M+1 Argininosuccinate _M+0 Argininosuccinate _M+1 Argininosuccinate _M+2 Argininosuccinate _M+3 Argininosuccinate _M+4 Arginine _M+0 Arginine _M+1 Arginine _M+2 Arginine _M+3 Ornithine _M+0 Ornithine _M+1 Ornithine _M+2 Carbamoyl_aspartate _M+0 Carbamoyl_aspartate _M+1 Carbamoyl_aspartate _M+2 Dihydroorotate _M+0 Dihydroorotate _M+1 Dihydroorotate _M+2 Orotate _M+0 Orotate _M+1 Orotate _M+2 UMP _M+0 UMP _M+1 UMP _M+2 UDP _M+0 UDP _M+1 UDP _M+2 UTP _M+0 UTP _M+1 UTP _M+2 Putrescine _M+0 Putrescine _M+1 Putrescine _M+2 METABOLITES_END #END