#METABOLOMICS WORKBENCH alanyl_20250326_080101 DATATRACK_ID:5781 STUDY_ID:ST003822 ANALYSIS_ID:AN007474 PROJECT_ID:PR002390 VERSION 1 CREATED_ON December 17, 2025, 1:18 pm #PROJECT PR:PROJECT_TITLE In vivo CRISPR screen identifies copper metabolism as a vulnerability in acute PR:PROJECT_TITLE lymphoblastic leukemia PR:PROJECT_SUMMARY The nutrient-sparse cerebrospinal fluid (CSF) poses a significant challenge to PR:PROJECT_SUMMARY spreading cancer cells. Despite this challenge, leukemia often spreads to the PR:PROJECT_SUMMARY CSF and represents a significant clinical complication. To uncover nutritional PR:PROJECT_SUMMARY dependencies of leukemia cells in the CSF that could be targeted PR:PROJECT_SUMMARY therapeutically, we conducted an in vivo targeted CRISPR screen in a xenograft PR:PROJECT_SUMMARY model of leukemia. We found that SLC31A1, the primary cell surface copper PR:PROJECT_SUMMARY importer, is a genetic dependency of leukemia in both the central nervous system PR:PROJECT_SUMMARY as well as in the hematopoietic organs. Perturbation of copper metabolism leads PR:PROJECT_SUMMARY to complex IV deficiency, perturbed nucleotide metabolism and slowed leukemia PR:PROJECT_SUMMARY cell proliferation. Furthermore, nutritional copper depletion reduced cancer PR:PROJECT_SUMMARY progression in cell line based and patient-derived xenograft models of leukemia. PR:PROJECT_SUMMARY Copper thus appears to be an actionable micronutrient in leukemia. PR:INSTITUTE Boston Children's Hospital PR:DEPARTMENT Pathology PR:LABORATORY Naama Kanarek PR:LAST_NAME Wong PR:FIRST_NAME Alan PR:ADDRESS 300 Longwood Avenue, Boston, MA, 02115, USA PR:EMAIL alan.wong@childrens.harvard.edu PR:PHONE (617) 355-7433 PR:FUNDING_SOURCE NCI 1R01CA282477-01A1 #STUDY ST:STUDY_TITLE Polar metabolite profiling of SLC31A1 knockout or control cells with or without ST:STUDY_TITLE rescue with over-expression of AOX ST:STUDY_SUMMARY To investigate whether the effects of SLC31A1 knockout can be rescued with ST:STUDY_SUMMARY alternative oxidase (AOX) overexpression, SEM cells containing a ST:STUDY_SUMMARY SLC31A1-targeting sgRNA (guide #2 or guide #7) or a intergenic -targetting sgRNA ST:STUDY_SUMMARY (i.e. a control sgRNA, "Intergenic1") and Cas9 were FACS-purified based on ST:STUDY_SUMMARY SLC31A1 expression, and then engineered to stably over-express either red ST:STUDY_SUMMARY fluorescent protein (RFP, as a control) or AOX from Ciona Intestinalis. Cells ST:STUDY_SUMMARY were then cultured in RPMI-1640 containing 10% FBS and penicillin and ST:STUDY_SUMMARY streptomycin in 6-well plates, and then collected for metabolomics. This ST:STUDY_SUMMARY demonstrated that the changes in aspartate, ureidosuccinic acid/carbamoyl ST:STUDY_SUMMARY aspartate and nucleotides were largely reverted by genetic overexpression of ST:STUDY_SUMMARY AOX. ST:INSTITUTE Boston Childrens Hospital ST:LAST_NAME Wong ST:FIRST_NAME Alan ST:ADDRESS 300 Longwood Avenue ST:EMAIL alan.wong@childrens.harvard.edu ST:PHONE (617) 355-7433 ST:NUM_GROUPS 4 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:CELL_STRAIN_DETAILS SEM leukemia cells #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 - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1309 Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1309.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1310 Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1310.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1311 Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1311.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1312 Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1312.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1313 Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1313.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1314 Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1314.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1315 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:RFP RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1315.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1316 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:RFP RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1316.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1317 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:RFP RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1317.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1318 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:AOX RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1318.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1319 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:AOX RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1319.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1320 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:AOX RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1320.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1321 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1321.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1322 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1322.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1323 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1323.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1324 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1324.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1325 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1325.raw SUBJECT_SAMPLE_FACTORS - 20241122_QE2_HILIC_KO_AOX_repeat_AYW1326 Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX RAW_FILE_NAME(raw file name)=20241122_QE2_HILIC_KO_AOX_repeat_AYW1326.raw #COLLECTION CO:COLLECTION_SUMMARY One and a half million cells from culture were collected via centrifugation for CO:COLLECTION_SUMMARY 20 seconds at 18,000xG at 4C, washed once with ice-cold 0.9% NaCl, and collected CO:COLLECTION_SUMMARY via centrifugation for 20 seconds at 18,000xG at 4C. Cells were cultured in CO:COLLECTION_SUMMARY RPMI-1640 with 10% FBS and penicillin/streptomycin in a 37C incubator with 5% CO:COLLECTION_SUMMARY CO2. CO:SAMPLE_TYPE Leukemia cells #TREATMENT TR:TREATMENT_SUMMARY Cells were seeded 0.5 million/mL into 4 mL of RPMI-1640 + 10%FBS + TR:TREATMENT_SUMMARY penicillin/streptomycin 48 hours prior to collection in 6 well plates. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Cell pellet was resuspended in 400uL of 100% LC-MS grade methanol supplemented SP:SAMPLEPREP_SUMMARY with isotopically-labelled amino acid standards [Cambridge Isotope Laboratories, SP:SAMPLEPREP_SUMMARY MSK-A2-1.2], aminopterin, and reduced glutathione standard [Cambridge Isotope SP:SAMPLEPREP_SUMMARY Laboratories, CNLM-6245-10]). with repeated pipetting up and down and vortexing SP:SAMPLEPREP_SUMMARY for 10 seconds. Then, 100uL of LCMS-grade water containing 125 mM Ammonium SP:SAMPLEPREP_SUMMARY Acetate, 10 mM Na-Ascorbate, and 7.9 mg/mL 5,5-dithio-bis-(2-nitrobenzoic acid SP:SAMPLEPREP_SUMMARY (Ellman's reagent) was added, and the sample vortex for another 10 seconds. SP:SAMPLEPREP_SUMMARY Samples were then centrifuged for 10 minutes at 18,000 g to pellet cell debris. SP:SAMPLEPREP_SUMMARY The supernatant was transferred to a new tube and dried on ice using a liquid SP:SAMPLEPREP_SUMMARY nitrogen dryer. The dried metabolites were then resuspended in 30uL and 2uL was SP:SAMPLEPREP_SUMMARY injected. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY 2 μL of each sample was injected into a ZIC-pHILIC 150 x 2.1 mm (5 μm particle CH:CHROMATOGRAPHY_SUMMARY size) column (EMD Millipore) operated on a Vanquish™ Flex UHPLC system (Thermo CH:CHROMATOGRAPHY_SUMMARY Fisher Scientific). Chromatographic separation was achieved using the following CH:CHROMATOGRAPHY_SUMMARY conditions: buffer A was 95% acetonitrile and 5% 20 mM ammonium carbonate, 0.1% CH:CHROMATOGRAPHY_SUMMARY ammonium hydroxide in water; buffer B was 95% 20 mM ammonium carbonate, 0.1% CH:CHROMATOGRAPHY_SUMMARY ammonium hydroxide in water and 5% acetonitrile. Gradient conditions used were: CH:CHROMATOGRAPHY_SUMMARY 0-20 min: linear gradient from 16.6% to 83.4% B; 20-24 min: hold at 83.4% B; CH:CHROMATOGRAPHY_SUMMARY 24-24.1 min: from 83.4% to 16.6% B; 24.1-32 min: hold at 20% B at 0.150 mL/min CH:CHROMATOGRAPHY_SUMMARY flow rate. The column oven and autosampler tray were held at 25 °C and 4 °C, CH:CHROMATOGRAPHY_SUMMARY respectively. CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME SeQuant ZIC-HILIC (150 x 2.1mm,5um) CH:SOLVENT_A 95% acetonitrile/5% water; 20mM ammonium carbonate; 0.1% ammonium hydroxide CH:SOLVENT_B 95% water/5% acetonitrile; 20mM ammonium carbonate; 0.1% ammonium hydroxide CH:FLOW_GRADIENT 0-20 min: linear gradient from 16.6% to 83.4% B; 20-24 min: hold at 83.4% B; CH:FLOW_GRADIENT 24-24.1 min: from 83.4% to 16.6% B; 24.1-32 min: hold at 20% B at 0.150 mL/min CH:FLOW_GRADIENT flow rate. The column oven and autosampler tray were held at 25 °C and 4 °C, CH:FLOW_GRADIENT respectively. CH:FLOW_RATE 0.15 mL/min CH:COLUMN_TEMPERATURE 25 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS MS data acquisition was performed using a QExactive benchtop orbitrap mass MS:MS_COMMENTS spectrometer equipped with an Ion Max source and a HESI II probe (Thermo Fisher MS:MS_COMMENTS Scientific) and polarity switching was used. Four scans were used: full scans in MS:MS_COMMENTS both positive and negative ionization mode in a range of m/z = 70–1000, with MS:MS_COMMENTS the resolution set at 70,000, the AGC target at 1 × 106, and the maximum MS:MS_COMMENTS injection time (Max IT) at 20 msec from 0-20 minutes. A third scan in the MS:MS_COMMENTS negative mode was used with range of m/z = 220-700 from 0-20 minutes and the MS:MS_COMMENTS same resolution, AGC settings with 30ms Max IT. Lastly, a targeted-SIM scan was MS:MS_COMMENTS added with a resolution of 35k, AGC target 1e5, and max IT 20ms, isolation MS:MS_COMMENTS window = 1.0 m/z, with an inclusion m/z of 503.0552 (corresponding to MS:MS_COMMENTS Ellman-derivatized glutathione). Tune file parameters were: spray voltage = MS:MS_COMMENTS 3.5kV, capillary temperature = 320C, S-lens RF = 50, auxillary gas temperature = MS:MS_COMMENTS 350C. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Normalized peak area MS_METABOLITE_DATA_START Samples 20241122_QE2_HILIC_KO_AOX_repeat_AYW1309 20241122_QE2_HILIC_KO_AOX_repeat_AYW1310 20241122_QE2_HILIC_KO_AOX_repeat_AYW1311 20241122_QE2_HILIC_KO_AOX_repeat_AYW1312 20241122_QE2_HILIC_KO_AOX_repeat_AYW1313 20241122_QE2_HILIC_KO_AOX_repeat_AYW1314 20241122_QE2_HILIC_KO_AOX_repeat_AYW1315 20241122_QE2_HILIC_KO_AOX_repeat_AYW1316 20241122_QE2_HILIC_KO_AOX_repeat_AYW1317 20241122_QE2_HILIC_KO_AOX_repeat_AYW1318 20241122_QE2_HILIC_KO_AOX_repeat_AYW1319 20241122_QE2_HILIC_KO_AOX_repeat_AYW1320 20241122_QE2_HILIC_KO_AOX_repeat_AYW1321 20241122_QE2_HILIC_KO_AOX_repeat_AYW1322 20241122_QE2_HILIC_KO_AOX_repeat_AYW1323 20241122_QE2_HILIC_KO_AOX_repeat_AYW1324 20241122_QE2_HILIC_KO_AOX_repeat_AYW1325 20241122_QE2_HILIC_KO_AOX_repeat_AYW1326 Factors Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP Sample source:SEM leukemia cells | guideRNA:Intergenic1 | overexpression:RFP Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:RFP Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:RFP Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:RFP Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:AOX Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:AOX Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #2 | overexpression:AOX Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX Sample source:SEM leukemia cells | guideRNA:SLC31A1 guide #7 | overexpression:AOX 2-hydroxyglutarate 2000664539 1283017741 1864108006 867271444.2 1399665216 1728102082 812338497.6 1192271812 1451213747 745254334.5 1292057228 1564022473 874132824.7 927318504.8 1466865287 1010343737 909047517.4 2235027769 2-oxobutyric_acid 31340974.1 31884617.73 33323685.21 34998589.56 35887920.62 28888509.5 30146557.23 29550394.91 27407954.23 28515100.53 37317347.46 33916951.21 35975918.76 36274141.84 35135915.2 38011598.78 52353971.04 39328874.6 3-phosphoglycerate 1529967.754 2406611.29 1884181.144 3545924.094 3061606.016 3221953.605 3759579.214 4607521.967 4074757.4 7395019.109 8915390.513 8733233.946 4916513.907 5180289.545 5566005.364 9417094.896 9455758.65 8044662.873 Acetyl.CoA 4352959.971 4677523.657 4552031.698 5704272.382 5022124.135 4479551.574 4658157.212 4775006.153 4459568.309 4930015.619 4517315.987 4755334.319 4566784.983 4612367.922 4838144.338 4701670.663 4621484.96 3710316.927 acetylcarnitine 51019608.13 58002273.98 55947078.08 53837307.38 52742116.53 53268764.07 59654667.81 56927187.88 57933638.31 55067770.15 55843349.98 55160476.56 60277035.23 59196235.38 60455264.99 57113755.66 50776079.34 48388629.78 adenine 34589773.43 22856346.56 40493756.79 14452375.81 41802372.79 44653005.53 58701751.72 42004072.62 29220672.84 28786080.46 29281320.6 26651002.8 24717084.82 29062106.42 42210020.53 23554803.18 18423928.13 26427969.06 ADP 457246826.8 504933310.8 452449411.8 531882799 504884089.4 454189549.2 384665149.6 369653745 381211891 447231101.3 437755553.2 445085477.2 415906382.4 399686797.8 341678180.6 462963738.6 432294743.2 325184310.5 alanine 87495438.5 84646037.51 83566882.26 81950478.1 81136914.79 79155017.9 91360389.08 80461510.6 86880406.82 71556455.23 61551562.38 65907027.4 90322581.31 84088063.19 87313084.42 63285281.89 66824005.91 67137275.22 AMP 74193759.3 52662780.18 87739714.99 46893568.79 95711625.45 93919670.14 111373620.4 108582922.9 92081768.47 105640351 71244845.79 57773225.95 71586663.59 91729742.83 103788219.6 43139793.86 71860823.78 132079851.7 arginine 1143614119 1158757550 1157733092 1296460895 1113158400 1160475924 1128821528 1067286581 1023101157 1166521814 1040288029 1041628098 1172006159 1188932195 1103134680 1131563891 1074175476 950169997.6 argininosuccinate 8034580.408 8186337.895 7721886.671 9653939.733 8828372.483 8533726.433 16299057.94 16058372.04 16298167.6 11288921.35 10380491.81 10048486.3 17331037.02 17533766.83 16816455.88 10464792.63 11349461.9 10206996.09 asparagine 677342362.3 637583574.4 635315289.3 619597526.6 600836089.7 616737438.3 481338382 474749031.5 499618694.1 537377775.2 533394003.5 521713373.8 489337356.1 460209402.3 467379454.8 550952285.5 555861267.7 581870085.8 aspartate 36084674.9 35140031.59 30551004.26 31539990.2 28536090.88 27156471.09 7195786.147 6471320.124 8031473.3 13334681.33 13014398.21 12427933.46 6361209.042 5643220.762 5162756.586 11503822.19 13035446.89 12253008.11 asymmetric.dimethylarginine 5613169.411 5895408.134 6011260.783 6559222.196 6045633.675 6231334.945 6729196.659 6593556.06 6078930.839 6541825.576 5868333.091 6020105.718 7033795.612 7083485.463 6624393.455 6123042.457 5469177.878 4774629.92 beta.alanine 16170007.8 24870720.42 20884247.93 19658546.9 25244521.45 22477185.2 31839799.11 32885014.31 29916509.22 26182992.54 23828976.55 22311085.79 33804939.58 35978482.91 33793053.48 27951425.12 21636934 22800386.37 Butyrylcarnitine 33549255.55 31935769.17 36722712.8 35833585.79 38268247.02 31774701.78 33180850 36925431.43 32776862.52 33159924.66 37030063.77 40716833.57 32497295.4 38558986.49 42310100.85 30233320.09 30813737.81 29987801.45 C5.Carnitines 4186700.841 3826023.582 3335739.472 3469726.367 2937149.351 2818849.992 10495086.78 10968636.72 9094305.875 2414827.324 3340613.498 3448625.232 11939410.67 11786633.3 12150409.82 2659275.57 2967750.007 2704847.897 carnitine 117991768.8 118764855.4 122109135.2 124286188.4 127158950 117402483.5 125892477.9 121173683.8 119873486.6 141262715.6 127755286.3 135619792.6 128990883.9 131943275 131933645.9 137685665.7 142288845.7 119294056.9 carnosine 907423.6635 1102692.239 1396634.919 999662.3641 1002735.527 1059541.94 1138079.554 1136206.775 987842.761 957393.1315 708258.3436 922697.961 918727.4203 1245956.143 974807.7557 658208.6909 765590.1051 883815.2754 CDP 39148963.5 38601872.58 44063449.98 43413998.95 48112212.2 44122986.59 31879605.77 28265847 31312348.89 46770583.54 38557442.63 37258214.03 29633680.99 28131499.87 25514389.15 36213455.18 39021126.16 37130483.38 cis.aconitate 20667176.31 25635696.63 17803100.31 28585934.64 17384389.57 19229794.65 16764904.08 15120605.92 26659694.22 27566789.27 19674338.77 26638620.67 19972889 26898241.85 15530186.33 20300700.1 28640007.41 25012479.15 citrate 209662877.3 227366862.7 189297899.3 288807874.4 229857684.2 225782312.1 250836087.8 241909474.3 229431998.1 308643929.8 303107703.9 259058054.4 240099248.6 264648706.6 227173643.6 268361948.8 306674694.5 273091960 citrulline 16872493.84 16218105.85 16295083.98 17081617.6 14659932.31 16432495.27 16215158.88 14839364.82 15907340.29 15443299.75 17033221.6 15453397.27 16020936.05 16240301.7 16157632.7 17104171.49 16285441.04 16751535.47 creatine 224351077.7 238475516 215228276.1 254347210.9 238379604.5 229618156.3 266242415.4 258279226.5 236572776.2 246429517 226890176.8 221498387.6 262081579.3 261925771.5 246290851.1 235722546.6 219978780 198531640.8 Cysteine_Ellmans_conjugated 11523736.02 11826754.84 12009828.91 9838230.085 10862146.76 15108537.86 11448901 9370667.719 9631716.548 10550076.94 7505996.856 7199005.051 11762582.48 9671308.207 10589033.16 9555320.646 6751498.79 8140550.431 cystathionine 5868219.861 5902993.43 5433228.521 6220093.493 5739373.545 5712572.599 4194864.871 4410228.62 5454308.191 4328949.542 4897534.751 5795487.988 4182665.694 4409341.696 5030361.464 4297374.22 5272253.489 5679158.432 dihydroxyacetone_phosphate 2627347.243 3521698.482 3012572.711 5034454.667 4802725.829 4267612.754 17884930.52 19962509.91 17241711.36 15055981.03 14648665.76 14426590.26 20617715.12 20625822.76 18839937.47 18201433.08 14374570.89 11267152.69 F6P_or_G1P 2163088.259 1794550.14 1867814.017 1960758.591 2156332.344 2095796.364 4217681.135 3729295.875 2893500.212 5713984.262 5754977.476 5328131.44 4505692.385 4145137.675 3750078.97 7770179.31 6239874.637 5352437.228 FAD 1130020.188 1388424.27 1165914.572 1308276.424 1421486.834 1193392.46 1234616.776 1278804.059 1377153.037 1481611.373 1420622.411 1210320.006 1225912.41 1611566.893 1061916.137 1559187.513 1255378.176 1302906.488 Fructose.1.6.bisphosphate 1030017.421 1100599.302 1085655.117 1164125.911 1129003.486 1111814.584 872694.5685 947076.1318 894131.0889 1820336.284 2089004.754 2089508.52 986030.705 918378.8103 924712.3876 2360421.077 1847340.428 1675373.004 fructose 41533358.3 41114443.62 34721394.99 37991685.73 37146841.43 35523988.39 33603705.68 35067364.6 34673116.11 38596362.6 40600990.7 41292857.5 36973477.16 37312869.34 33983240.87 39780670.11 46693977.02 47685363.76 fumarate 13916905.47 19876312.76 14068053.49 24184054.29 18116108.39 22813576.57 28259384.19 28657651.54 29852113.34 22139667.72 18703310.28 16540637.28 31553964.15 30198541.96 31725486.27 17333191.8 19211601.02 15217998.39 GDP 53164441.05 58128873.63 57841878.76 68118402.01 57213991.23 53138867.5 65598518.71 63312458.77 61789663.95 63852387.13 65217228.67 68380265.77 81446081.47 72290689.22 61812984.95 76529055.65 67667756.6 48143343.25 glutamate 1093243948 1135589211 1028945842 1174030657 1072340697 988465610.1 695976173.5 623790984.6 744647492.6 850468616.4 819753077.7 843739990.1 689020174.8 597780670.9 577942842.1 831115770.9 873911505.1 753139839.3 glutamine 2384737268 2316995868 2247664264 2231406296 2197709578 2214712700 1219482242 1059334252 1166687868 1638896215 1392981184 1407096719 1063958487 975194953 986317675 1498794727 1505151428 1473372745 Glycerol.3.Phosphate 9646870.861 9947124.374 10623319.88 10036851.05 10658050.17 10277740.54 36212522.5 39988898.54 38154582.39 25465363.03 26200321.34 25225370.06 40407439.03 41303197.08 43301252.78 31026349.26 28212212.96 28087805.18 Glycerophosphocholine 1969745183 2019005486 2027907933 2149740477 2040374459 2134570733 1157403466 1151457979 1272781826 1796976879 1770665635 1887187533 1070962668 1091220011 1221083788 1622500863 1784763817 1852783776 glycine 9848759.082 10363572.75 10083062.37 10515376.49 10942062.57 10938022.36 8059780.605 7659273.783 7942374.387 10334723.78 10573324.78 8562121.175 8422769.249 7807905.342 7556238.295 9130181.332 10908773.35 9954444.865 GMP 10799174.48 7924623.407 11922131.85 6876831.449 15093381.18 11561526.11 21516517.17 21388869.01 17351957.4 18515140.84 12231789.27 11383258.87 15871125.64 19148320.51 20581688.21 8211363.623 13673788.4 22727784.61 GSH_Ellmans 274326205.8 297141482.9 303572847 342823830.8 308008053.5 277131038 261205969.1 267467861.5 262435854.6 304181159.1 299097821.8 286177981.9 267260242.5 270404164.5 257005535.9 304010241.3 303424938.8 249833855 GSSG 3990029.853 4607382.084 4320918.399 4149694.716 5199140.939 5401229.304 3912863.35 4460951.382 4233182.011 4285498.997 4497732.104 4694109.398 4048062.506 4156502.847 4753020.29 5377205.702 3639372.76 3623365.38 histidine 255805436.3 233359817.9 236499403.6 232034189.4 246206388.1 243783045.4 231240606.8 220365860.5 212775186.8 237235414.8 212598186.4 195051574.5 216462808.6 211555696.2 205681093.1 203620269.9 218321770.6 216530542.3 homocitrulline 1318837.934 1170957.342 914498.0667 1129197.323 1078765.669 1034878.205 1600225.25 1537078.093 1316651.029 1539122.896 911403.3037 1278343.806 1465505.612 1319880.364 1312443.54 1131785.522 1012355.758 1246089.96 hypotaurine 60724620.62 32175077.44 53273997.66 25130572.87 33611695.93 49086645.95 30523500.26 40016808.74 46782484.36 24838826.24 37213963.07 43816879.83 30918832.15 25948183.86 46422931.91 32564732.66 26260384.32 44381331.48 IMP 629343.9253 662608.0156 713635.4405 885968.9825 839022.8915 769568.9218 6672349.469 7700214.663 5821940.7 3719930.964 4574503.672 4327423.825 9076959.295 9248313.804 8022281.963 4902777.554 4367928.528 3363229.243 isoleucine 322843791.1 374190742.6 347803314.3 371168254.6 377091995.3 379855786.3 385331363.2 294401531 337251322.1 334607833 291442313.8 278116652.7 378760694.2 254005033.3 279165536.3 316194061.4 291470479.1 280022609.6 kynurenine 2489033.303 3262343.351 2345499.247 2858765.604 2454980.613 2630241.518 3793775.816 3564415.32 3774772.822 2597439.832 2635685.254 2673909.126 3525791.225 3396701.653 3356977.84 2591512.8 3544134.089 2726026.599 leucine 251789240.6 280517345.9 221240522 264358086.8 213439004.6 230723776.2 252770320.2 217212438.8 230482064.7 219279336.1 230305025.2 226355147.3 192962231.7 208183716.9 243666901 202148010.4 205614957 240954819.3 lysine 11705702.88 12930002.98 11775416.58 12586117.72 10933082.93 11541038.37 10272167.97 10368040.68 9875603.809 11095765.58 9632632.114 9295019.104 10961096.27 11105718.96 10740769.07 9470637.81 10195810.55 8990995.855 malate 236928273.8 272450717.9 233409615.5 291021285.2 281435528.2 255982519.3 410156110 377917677.9 388116755.6 304958196 278866397.8 252536910.4 420743739.6 402128238.4 371921371.4 305415009.3 308350339.5 253140900.3 mannose 41215463.23 41109367.54 34721032.69 37988343.58 37149089.25 35631635.52 33425409 35068746.8 34762290.98 38599461.89 40262349.22 41318215.68 36752834.95 37133355.51 33803948.22 39993091.63 46580780.36 47686619.58 Methionine.sulfoxide 70807752.98 72781549.59 74257020.82 71544885.57 73436254.84 76347368.15 66532282.99 64530135.99 64533907.82 67508940.02 60421984.8 62857748.19 61977629.57 60245081.35 61772341.73 66662707.06 57651670.63 59045872.06 methionine 136405409.5 137550627.5 126657089.6 126000073.3 122317582.8 149038166.9 109121303.2 88380250.62 95667696.69 113800162.9 91207442.23 85584888.69 112626977.3 91753853.28 86921274.19 114523223.1 104548803.6 101569838 methylthioadenosine 2918875.985 2874877.155 3347564.128 3486247.617 3666083.708 3345391.751 3371779.027 3551881.566 3220258.61 2894880.369 3919788.85 4345414.538 2704547.126 3394169.159 3541300.975 3270013.069 2985448.138 3434379.938 NAD. 83470097.07 86410265.67 86661803.96 93055244.93 90300763.26 84942308.7 74684785.94 77604641.63 76072492.81 84502195.99 85947838.33 86844918.25 76884452.33 78696181.57 74207471.8 86010116.77 85693597.67 79073094.81 NADH 11928056.59 12743422.88 13204224.24 12782629.09 11885569 11759905.63 19828054.8 24546968.77 23732405.27 16749983.28 17973989.28 17242538.26 23084374.14 24857937.01 24969785.31 20945786.52 17940532.83 15870603.59 NADP 700283.3455 882228.0816 736753.8851 858714.1495 866099.1859 799930.2694 752974.8541 830334.5434 711578.1139 865003.1749 592024.0172 692186.7429 704354.4959 822807.6278 760597.8692 754481.1783 581337.7816 683155.6959 NADPH 4410542.529 4758784.44 4373470.966 4954500.448 4526758.322 4240101.904 3725180.969 3737760.282 4080453.442 3822113.74 3823480.445 4153771.131 3727990.708 3818425.209 3580639.878 4086784.092 3636841.636 3561861.39 niacinamide 12757978.83 14089800.12 15440660.38 16947868.68 16025783.53 17001741.52 11651893.04 13171320.18 9178194.242 12965004.24 14849538.52 17606143.09 9710851.633 15430744.23 11546675.35 15910979.94 22558404.49 16854922.03 ornithine 14387001.78 15350520.64 15086962.74 16547385.23 14248640.37 15867147.75 12859280.28 12573350.73 12229479.44 14127187.52 13290891.82 12571936.42 14882109.62 14285743.99 12612865.59 11562054.22 12820627.01 12726511.99 palmitate 145116109.6 76168970.92 111844621.3 107974587.8 65092485.16 60752612.53 52272835.32 65439144.21 77545580.23 49960430.94 73637379.12 101266344.2 75515900.37 74711274.18 63282124.72 100028954.6 170277084.6 195717912.9 phosphocholine 11485500468 11252298135 11610130056 11524864426 12259634967 12144328285 8144121640 9069207879 9415545603 11004375748 11596847199 11503640135 7942909082 8181474643 8949972725 10483469354 10649535226 11316510356 phosphocreatine 1224086.019 1293114.935 1333871.095 1228250.316 990786.3368 1108429.83 1116045.886 831708.3914 1089117.597 1070171.012 855407.0285 810029.958 904830.8546 763271.01 964555.8867 1068791.873 759363.4419 739135.0108 phosphoenolpyruvate 476652.0903 330855.7042 453670.3318 538133.947 628714.8168 735155.5622 470544.9758 468514.5425 505588.1385 1508755.775 1377394.695 1750379.413 464778.1981 561189.0058 524739.874 1673697.669 1598484.419 1289893.746 proline 2254205405 2366499089 2079671157 2476188041 2508486771 2443896168 3069029723 2685321504 2773822650 2967501481 2625037002 2886845480 3083970146 2777637974 2556643111 3057677690 2932016668 2900140488 propionylcarnitine 2704495.045 2939959.303 2350232.175 3123614.038 2980769.424 2678737.169 1321496.897 1395348.339 862843.5906 2475754.115 2531499.809 2610875.838 1454487.306 1453473.464 1663978.94 2274299.379 2225772.212 1923410.149 Pyridoxine 32774451.69 31118951.14 34238244.57 33527165.8 35095498.99 34106942.91 30496761.21 30285059.1 31864934.33 36487758.98 34248881.53 38023346.28 33387199.64 35943885.95 32975989.33 32051937.78 30003470.66 31071871.76 S.Adenosyl.L.Homocysteine 4048132.889 4600092.668 4747603.165 4673317.674 5247215.913 4639803.855 3462601.556 4797457.502 4751597.804 3722743.9 5154717.853 4567240.043 3900453.051 3725051.845 4378689.315 4776960.269 3671739.566 4672789.708 S.Adenosyl.L.methionine 55504450.77 58254909.69 55259253.73 64269499.9 61562658.31 56982833.67 55914591.36 54271180.51 57023348.25 61181064.22 63170074.58 57298596.1 55419379.15 58883027.25 52488741.43 54781299.26 69300476.15 59129366.98 sarcosine 90321837.48 84489432.27 83595433.06 80930650.4 78490350.18 79005145.31 91994572.87 81246968.54 85177420.14 71488400.45 59586040.87 64521776.28 87705613.76 84345439.14 88028091.14 61204243.79 66512473.76 66432562.83 Sedoheptulose.7.phosphate 1588789.249 1659236.758 1657372.796 1896793.646 1788154.107 1255934.334 4276902.272 4559558.385 3475697.585 5659470.818 4787588.07 4997102.56 3577897.43 4014381.843 3613758.522 7847125.295 5636804.386 4336856.026 serine 40554326.56 42606481.09 44940707.44 32310360.11 37058458.16 36923156.55 21934433.13 22081458.58 23991192.07 22077188.2 25788802.95 23300053.51 23484903.12 21163544.22 22497148.03 22621869.37 26241472.44 32145756.48 succinate 45838636 38558937.91 38815037.66 25944746.3 41910810.67 48178738.3 64036565.44 60465136.79 75468728.86 33030964.06 38683780.42 40611087.58 78929502.55 58712609.48 76921708.54 43118513.46 33365543.1 62387086.95 succinyl.CoA 842934.187 1006140.463 710814.7298 1104143.581 828713.8775 795306.8055 113857.8759 162247.6947 228375.619 556631.8073 690598.2635 941116.0185 253920.2674 205215.9947 148379.1637 759515.3198 666200.3254 502081.6135 taurine 241620738.7 243832006.9 237852877.4 261879174 243301965.4 235873110.1 269992930.5 270511290.9 278159178.8 264174765 244088919.2 264440702.4 282993623 278492329.1 270951632.6 281968222 249544211.7 222655412.6 threonine 219625063.2 156533413.7 193414387.8 141558693.1 160332107.7 180312254.2 55421849.22 62938732.43 81314682.33 69076525.53 65479558.87 81194903.58 53016447.57 58507264.67 65782001.95 73603120.59 63362438.43 88849353.67 trans.urocanate 15865955.21 21506730.01 26203515.14 24653462.98 10832415.22 12899294.18 8236987.32 13604131.37 9444006.971 8980911.543 41011845.36 14759547.02 13604782.96 7199904.294 7109647.897 9467162.54 12626847.2 8637397.895 tryptophan 41511875.5 47144930.74 40718629.94 42465037.47 41947897.98 49014497.59 39036639.25 33194790.94 33471025.54 39132397.88 32989503.82 30016408.15 42209839.72 35114208.85 33820805.24 40514624.69 39807284.32 33233541.14 tyrosine 224980160.2 179148671.5 212571051.7 154421489.5 179120497.4 217292805 117707945.2 135516448.4 149635325.3 122078871.1 132383581.4 151433217.8 119678493.3 113096016.6 136478111.1 136246953.8 130415317.8 161503289.7 UDP.GlcNAc 46860624.61 47338226.42 46214720.79 49469996.18 47327099.77 45411857.75 30028588 30676757.16 31632571.26 35992821.69 32715410.84 35261398.19 26130866.27 27832859.91 27757280.34 33551753.17 33565794.63 31733380.16 UDP 135880930.7 133913643.3 139017410.2 132239681.2 156506048 140407001.7 102811650.9 96158105.44 100507993.8 140759348.1 120070820.7 120168768.7 92910046.12 89651252.77 82571669.85 118331538 125007719.1 115452158.3 UMP 12349173.17 8868307.501 14839241.5 7429289.476 20389776.77 15460137.53 17048575.43 15189312.94 12940532.43 21303645.56 11896392.7 9041484.409 9715459.779 13269988.36 12896048.32 6720982.711 13267239.6 21981556.71 Carbamoyl Aspartic Acid 5235899.996 4381636.911 5156996.985 NA NA NA 18363079.1 17025481.72 22829039.85 NA NA NA 15859576.54 13886256.24 16136237.41 NA NA NA valine 82819524.37 82539122.26 70759829.86 54518001.14 78005171.16 84981946.28 48141441.36 70879761.99 84231201.2 45785431.33 67216046.09 81330200.31 41655344.1 53206491.88 65965316.92 54732234.19 59257840.52 83275565.49 xanthosine 3887510.611 4485383.318 3854709.751 4526282.477 4115603.743 3897672.521 4074800.727 3804575.648 4115000.697 4012728.12 5568816.15 4435902.888 4527555.444 3700513.563 3913355.591 3887054.808 4865267.209 5560375.571 dihydroorotate 569096.9671 754418.41 423668.1059 46434.90312 66972.56264 39582.99267 1850812.483 1794435.414 3133724.074 0 0 0 1985850.166 1647257.949 1740886.423 0 0 81054.04217 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name 2-hydroxyglutarate 2-oxobutyric_acid 3-phosphoglycerate Acetyl.CoA acetylcarnitine adenine ADP alanine AMP arginine argininosuccinate asparagine aspartate asymmetric.dimethylarginine beta.alanine Butyrylcarnitine C5.Carnitines carnitine carnosine CDP cis.aconitate citrate citrulline creatine Cysteine_Ellmans_conjugated cystathionine dihydroxyacetone_phosphate F6P_or_G1P FAD Fructose.1.6.bisphosphate fructose fumarate GDP glutamate glutamine Glycerol.3.Phosphate Glycerophosphocholine glycine GMP GSH_Ellmans GSSG histidine homocitrulline hypotaurine IMP isoleucine kynurenine leucine lysine malate mannose Methionine.sulfoxide methionine methylthioadenosine NAD. NADH NADP NADPH niacinamide ornithine palmitate phosphocholine phosphocreatine phosphoenolpyruvate proline propionylcarnitine Pyridoxine S.Adenosyl.L.Homocysteine S.Adenosyl.L.methionine sarcosine Sedoheptulose.7.phosphate serine succinate succinyl.CoA taurine threonine trans.urocanate tryptophan tyrosine UDP.GlcNAc UDP UMP Carbamoyl Aspartic Acid valine xanthosine dihydroorotate METABOLITES_END #END