#METABOLOMICS WORKBENCH alanyl_20251204_065215 DATATRACK_ID:6836 STUDY_ID:ST004464 ANALYSIS_ID:AN007482 PROJECT_ID:PR002390 VERSION 1 CREATED_ON December 17, 2025, 3:09 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 SEM-luciferase cells purified from the skulls and ST:STUDY_TITLE spleens of mice on copper replete diet (CuR+vehicle) or copper depleted diet ST:STUDY_TITLE with zinc acetate drinking water (CuD+ZnAc) and pulsed with 3-13C serine tracer. ST:STUDY_SUMMARY This study aimed to measure differences in purine synthesis between leukemia ST:STUDY_SUMMARY cells from the central nervous system (CNS) and spleens of mice, and also to ST:STUDY_SUMMARY determine the effect of copper depletion on purine synthesis in these two sites. ST:STUDY_SUMMARY Mice were irradiated, engrafted with leukemia cells and randomized to either ST:STUDY_SUMMARY CuR+vehicle or CuD+ZnAc treatments. 16 hours prior to harvest, mice were dosed ST:STUDY_SUMMARY with 800mg/kg 3-13C serine by intraperitoneal injection. Leukemia cells were ST:STUDY_SUMMARY harvested from the CNS and spleens of mice using magnet-activated cell sorting ST:STUDY_SUMMARY (MACS). This experiment revealed accumulation of purine synthesis intermediates ST:STUDY_SUMMARY in CNS leukemia cells compared to splenic leukemia cells, and that copper ST:STUDY_SUMMARY depletion leads to reduced label incorporation into purines. 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 #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 - 20251030_QE1_HILIC_SEMserineMACS_AYW1357 Sample source:CNS | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1357.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1358 Sample source:Spleen | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1358.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1359 Sample source:CNS | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1359.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1360 Sample source:Spleen | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1360.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1361 Sample source:CNS | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1361.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1362 Sample source:Spleen | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1362.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1363 Sample source:CNS | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1363.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1364 Sample source:Spleen | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1364.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1365 Sample source:CNS | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1365.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1366 Sample source:Spleen | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1366.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1367 Sample source:CNS | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1367.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1368 Sample source:Spleen | treatment:CuR+Vehicle RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1368.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1369 Sample source:CNS | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1369.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1370 Sample source:Spleen | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1370.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1371 Sample source:CNS | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1371.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1372 Sample source:Spleen | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1372.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1373 Sample source:CNS | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1373.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1374 Sample source:Spleen | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1374.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1375 Sample source:CNS | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1375.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1376 Sample source:Spleen | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1376.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1377 Sample source:CNS | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1377.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1378 Sample source:Spleen | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1378.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1379 Sample source:CNS | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1379.raw SUBJECT_SAMPLE_FACTORS - 20251030_QE1_HILIC_SEMserineMACS_AYW1380 Sample source:Spleen | treatment:CuD+ZnAc RAW_FILE_NAME(raw file name)=20251030_QE1_HILIC_SEMserineMACS_AYW1380.raw #COLLECTION CO:COLLECTION_SUMMARY Mice were anesthetized with ketamine/xylazine (100-120mg/kg; and 10mg/kg by CO:COLLECTION_SUMMARY intraperitoneal injection), and cells were harvested from the skulls and spleens CO:COLLECTION_SUMMARY of mice by blunt dissection into 1xPBS with 2% FBS and 2mM EDTA on ice (PBE). CO:COLLECTION_SUMMARY Cells were pelleted, then resuspended in 160uL of PBE, and 50uL of Mouse Cell CO:COLLECTION_SUMMARY Depletion Cocktail (Miltenyi Biotec cat #130-104-694) was added per sample and CO:COLLECTION_SUMMARY samples incubated on ice for 5 minutes. While incubating, one LS column CO:COLLECTION_SUMMARY (Miltenyi Biotec 130-042-401) per sample was affixed to a QuadroMACS magnet CO:COLLECTION_SUMMARY (Miltenyi Biotec 130-091-051) and primed using 3mL of ice-cold PBE. The cell CO:COLLECTION_SUMMARY suspension was then applied to each column and the flow-through containing CO:COLLECTION_SUMMARY unlabelled, human leukemia cell-enriched fraction was collected. The column was CO:COLLECTION_SUMMARY washed with an additional 2x1mL of PBE and the flow-through collected as well. CO:COLLECTION_SUMMARY The columns were then discarded. The flow-through was spun at 4000g for 1 minute CO:COLLECTION_SUMMARY at 4C, and the cell pellet resuspended in 1mL of 0.9% NaCl. After an additional CO:COLLECTION_SUMMARY 20 second spin at 4C and 21,000rcf, pelleted cells were resuspended in 100% CO:COLLECTION_SUMMARY LC-MS grade methanol for metabolite detection. CO:SAMPLE_TYPE Leukemia cells #TREATMENT TR:TREATMENT_SUMMARY NOD-SCID mice (5 weeks or older) were randomized by cage to either a copper TR:TREATMENT_SUMMARY replete diet (6ppm, Teklad TD.240097) with 2% sucrose in the drinking water TR:TREATMENT_SUMMARY (SigmaAldrich S0389-1kg) or copper deficient diet (0.3-0.7ppm, Teklad TD.240098) TR:TREATMENT_SUMMARY with 2% sucrose and zinc acetate (SigmaAldrich 383317-25g) in the drinking TR:TREATMENT_SUMMARY water. The zinc acetate dose was 2g/L for 4 weeks (2 weeks prior to injection TR:TREATMENT_SUMMARY and 2 weeks after); after this treatment period, all mice were switched to TR:TREATMENT_SUMMARY autoclaved water. Mice were allowed to consume food and water ad libitum, and TR:TREATMENT_SUMMARY the food and water was replaced every week. Two weeks after starting these TR:TREATMENT_SUMMARY treatments, mice were irradiated (250 rads) and then engrafted with 3 million TR:TREATMENT_SUMMARY SEM-luciferase cells in 200uL sterile 0.9% NaCl by tail vein injection. On the TR:TREATMENT_SUMMARY 25th day after injection, 16 hours prior to euthanasia, all mice were dosed with TR:TREATMENT_SUMMARY 800mg/kg 3-13C labelled serine in sterile 0.9% NaCl by intraperitoneal TR:TREATMENT_SUMMARY injection. #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 0-17.5 min: linear gradient from 16.6% to 75.0% B; 17.5-18.0 min: linear CH:CHROMATOGRAPHY_SUMMARY gradient from 75.0% to 85.0% B; 18-20.0 min: hold at 85.0% B; 20.0-20.5 min: CH:CHROMATOGRAPHY_SUMMARY from 85.0% to 16.6% B; 20.5-24 min: hold at 16.6% B at 0.150 mL/min flow rate. CH:CHROMATOGRAPHY_SUMMARY 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 0-17.5 min: linear gradient from 16.6% to 75.0% B; 17.5-18.0 min: linear CH:FLOW_GRADIENT gradient from 75.0% to 85.0% B; 18-20.0 min: hold at 85.0% B; 20.0-20.5 min: CH:FLOW_GRADIENT from 85.0% to 16.6% B; 20.5-24 min: hold at 16.6% B 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. Two 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 × 10e6, and the maximum MS:MS_COMMENTS injection time (Max IT) at 20 msec from 0-20 minutes. Tune file parameters were: MS:MS_COMMENTS spray voltage = 3.5kV, capillary temperature = 320C, S-lens RF = 50, auxillary MS:MS_COMMENTS gas temperature = 350C. Targeted TraceFinder analysis for 13C-labelled MS:MS_COMMENTS metabolites. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area MS_METABOLITE_DATA_START Samples 20251030_QE1_HILIC_SEMserineMACS_AYW1357 20251030_QE1_HILIC_SEMserineMACS_AYW1358 20251030_QE1_HILIC_SEMserineMACS_AYW1359 20251030_QE1_HILIC_SEMserineMACS_AYW1360 20251030_QE1_HILIC_SEMserineMACS_AYW1361 20251030_QE1_HILIC_SEMserineMACS_AYW1362 20251030_QE1_HILIC_SEMserineMACS_AYW1363 20251030_QE1_HILIC_SEMserineMACS_AYW1364 20251030_QE1_HILIC_SEMserineMACS_AYW1365 20251030_QE1_HILIC_SEMserineMACS_AYW1366 20251030_QE1_HILIC_SEMserineMACS_AYW1367 20251030_QE1_HILIC_SEMserineMACS_AYW1368 20251030_QE1_HILIC_SEMserineMACS_AYW1369 20251030_QE1_HILIC_SEMserineMACS_AYW1370 20251030_QE1_HILIC_SEMserineMACS_AYW1371 20251030_QE1_HILIC_SEMserineMACS_AYW1372 20251030_QE1_HILIC_SEMserineMACS_AYW1373 20251030_QE1_HILIC_SEMserineMACS_AYW1374 20251030_QE1_HILIC_SEMserineMACS_AYW1375 20251030_QE1_HILIC_SEMserineMACS_AYW1376 20251030_QE1_HILIC_SEMserineMACS_AYW1377 20251030_QE1_HILIC_SEMserineMACS_AYW1378 20251030_QE1_HILIC_SEMserineMACS_AYW1379 20251030_QE1_HILIC_SEMserineMACS_AYW1380 Factors Sample source:CNS | treatment:CuR+Vehicle Sample source:Spleen | treatment:CuR+Vehicle Sample source:CNS | treatment:CuR+Vehicle Sample source:Spleen | treatment:CuR+Vehicle Sample source:CNS | treatment:CuR+Vehicle Sample source:Spleen | treatment:CuR+Vehicle Sample source:CNS | treatment:CuR+Vehicle Sample source:Spleen | treatment:CuR+Vehicle Sample source:CNS | treatment:CuR+Vehicle Sample source:Spleen | treatment:CuR+Vehicle Sample source:CNS | treatment:CuR+Vehicle Sample source:Spleen | treatment:CuR+Vehicle Sample source:CNS | treatment:CuD+ZnAc Sample source:Spleen | treatment:CuD+ZnAc Sample source:CNS | treatment:CuD+ZnAc Sample source:Spleen | treatment:CuD+ZnAc Sample source:CNS | treatment:CuD+ZnAc Sample source:Spleen | treatment:CuD+ZnAc Sample source:CNS | treatment:CuD+ZnAc Sample source:Spleen | treatment:CuD+ZnAc Sample source:CNS | treatment:CuD+ZnAc Sample source:Spleen | treatment:CuD+ZnAc Sample source:CNS | treatment:CuD+ZnAc Sample source:Spleen | treatment:CuD+ZnAc adenine_C0 0 0 0 0 0 0 0 0 0 0 0 0 13853376 0 0 0 0 0 0 0 0 0 0 0 adenine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 adenine_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 adenine_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 adenosine_C0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 adenosine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 adenosine_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 adenosine_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ADP_C0 981054518 2616685096 1994062443 1724536545 3208529723 461789476 2942952447 142260192 3095336678 2796775000 2600838691 1247856442 813311307 29368144 289081188 112126544 3564978560 691607440 3014526531 940776497 3442791549 788856174 2100017906 112817984 ADP_C1 179103013 414120714 326890230 258077705 536598892 74648829 505163715 26642018 531489407 451867705 481464697 207333605 142219750 420678 48697937 16447429 572871687 90247487 500822021 124753892 550365609 104932918 334506373 10470814 ADP_C2 74985662 146635867 142279553 85227517 233058409 26944871 237096415 8895562 233854798 157064274 179340316 67609343 40015635 201985 18836442 3883603 230095140 25512318 192237526 38836997 216249650 33102375 121419761 3004027 ADP_C3 8777995 16944914 16536457 7852361 28653264 1301755 33403984 337496 31768808 17466281 26396818 7744121 2441693 0 637793 0 24930301 1104730 21144234 2405831 26406335 1241022 12729971 196227 AICAR_C0 128144998 6598664 208969263 33140083 225524235 1082481 181673813 0 205430478 5170140 263721400 2349682 28461995 0 8294886 0 199244474 0 226582405 376538 356572899 0 269196209 0 AICAR_C1 14729571 0 22616624 3105075 26640991 0 20974551 0 23698722 0 29310969 0 1936489 0 0 0 19500881 0 24944238 0 37681264 0 29473479 0 AICAR_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 AMP_C0 421753107 1157444965 792632363 940845759 1558240885 242474980 1724760187 47979855 1404555349 1318252747 1269057159 582256407 442064561 9407341 139990959 34438783 2194930117 297035896 1622501792 443473715 1688734678 318140481 1230859510 37313536 AMP_C1 71223097 177277852 119181397 130465617 241491158 35609354 286834252 7075079 225450581 204413798 213921861 87034556 71878855 1006469 20573744 3688804 326936649 29540387 254346940 57227316 253441595 39289744 189103413 4689300 AMP_C2 53478557 67507461 59975400 55335435 88886075 39161953 114444065 38628196 89360161 69823904 90261639 51830615 45758531 32360674 44452635 37674138 105646335 41767731 93182226 42210933 86901585 43222764 78863198 34836505 AMP_C3 0 0 0 0 0 0 1047778 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ATP_C0 1225997436 5273189240 2982491348 1311958936 4441247521 407676269 3877401074 354402829 4880041867 4250006361 3808938741 1455030379 354392889 43870491 143499975 217780413 3767006821 1183024086 3382751892 935999448 4542385947 1255883895 1020786347 119125591 ATP_C1 239076880 833988707 492860434 205106500 762840265 66497740 681944253 65268775 857271989 697295227 717956901 243196721 65322000 6623813 24909496 33295657 609131599 152275603 572132267 133601333 754927603 171974989 172035536 16481906 ATP_C2 90210998 278112444 156610093 64713517 244972235 21444508 247601837 22323321 298691495 228362944 252169966 80878366 14938826 937949 8243352 9991641 201933023 43612927 191719228 36655107 249022928 50145693 54851474 4382808 ATP_C3 12601516 36729395 21186800 8099441 35628591 2354503 39719528 2309137 44948786 32705378 43864826 11358005 1354774 0 517156 496946 25102183 4159641 24897571 3827941 31615113 4871245 6741689 202255 dATP_C0 925395 0 0 11095288 0 0 0 0 0 0 0 0 0 0 6096394 0 0 0 0 10050994 0 0 18831418 0 dATP_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 dATP_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 dATP_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 dimethylglycine_C0 58494103 149864377 145697876 153153787 172104488 59192741 439439800 46033580 189585511 234591521 327559141 221187765 48603660 17493145 28634007 22543607 131150807 31227946 89331014 46334612 138706497 49818346 47107140 12021919 dimethylglycine_C1 2436793 6886546 7645076 6316206 8478161 2627298 20421429 2695411 11607553 11279817 16779129 9430204 2403017 915691 1575205 1055503 5763130 1983457 3707095 2129738 6765335 2525602 4296625 717852 dimethylglycine_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 dTMP_C0 82369477 200874935 150359132 121039613 222001095 92513526 210570548 87056236 199210775 204696560 188802727 122035032 101093627 81028926 97633174 99013159 213776352 110133098 194406222 94937034 203229231 97900266 139225328 98381600 dTMP_C1 6479472 21095814 12093166 10030735 23328331 4505885 18758378 4057264 15768605 20790553 18160601 11415063 4399386 2347432 3869643 3552966 19690902 3115889 15091722 2900594 18693175 3654087 9380185 2768800 dTMP_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 dTMP_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 dTTP_C0 20153309 122421021 58986849 16625553 87655734 3560539 78202408 6400508 107376471 83496396 84292773 16166725 3850202 1695999 1333965 2802676 76334816 16080081 72150324 6718397 95887694 9291982 16524944 2433854 dTTP_C1 2107127 16347828 7464933 1682857 9915691 0 9967688 78596 12575158 11433874 10815364 1722053 0 0 0 0 9222667 1119084 9361488 115607 11302030 308296 1016979 0 dTTP_C2 83689 3602289 1092181 131583 2446484 0 1126622 0 3216899 754385 1662876 110032 0 0 0 0 1568637 91738 1611554 0 2294719 0 0 0 dTTP_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 formyl_methionine_C0 1970838 12848131 6687453 7479003 16803244 1522627 15055124 0 14702748 17878543 11157756 9951351 2058190 0 0 0 16432907 254699 11821600 2920019 15742898 678332 10527614 0 formyl_methionine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 GAR_C0 16926942 119909 27607803 436544 34749048 0 26795860 0 26087588 0 26799932 0 535078 0 0 0 23025057 0 29538376 0 47996129 0 49399094 0 GAR_C1 0 0 308691 0 640871 0 559386 0 257914 0 326399 0 0 0 0 0 0 0 316116 0 1842916 0 2863963 0 GDP_C0 113313364 223338487 231039189 155878552 375095722 48653587 349086923 14809378 325812442 251813834 286326476 122261032 79941985 2552052 31030385 9104070 394895070 61975208 315316799 79872998 358384745 65428863 219643101 10097143 GDP_C1 16586345 31154909 31999672 20463784 54787251 5830575 53396985 1277786 50540456 35877415 42797558 16868712 12508541 0 3469454 624294 57492080 6187813 47412989 9706429 51526300 6980152 30766618 278649 GDP_C2 4452979 8528818 8225067 5332442 12977909 1184216 13545191 71655 11564105 10078011 12924444 4400493 1794407 0 526115 73410 13469436 1856349 10895623 1499834 12224182 902600 7987140 0 GDP_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 glycine_C0 80017054 176761329 168645480 128229427 261314994 75594072 245822556 26079546 188662151 189846013 174934549 115835030 63762092 9885352 42778475 22775824 177123353 49140143 164708032 82221551 173965124 65147692 135995573 16753783 glycine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 GMP_C0 80749707 214739546 274222586 139653440 374971603 33693950 547719623 6692001 424947030 199871890 247627553 75366189 76670087 1577643 26169669 4891092 584450924 44753789 378253105 55215085 490976609 35175355 200403299 5201198 GMP_C1 12134843 29817758 35010592 17931783 51064466 2627110 74823869 2406456 57067669 28286853 39276025 6630156 10299824 1114267 4266264 910822 77515070 5751891 50852899 4236769 63560213 2466400 24251900 424624 GMP_C2 2200145 6417638 6823262 3778180 10770246 556336 16817733 0 11795657 6028411 8039939 2426388 532355 65165 379261 80386 15245254 427062 10087876 716859 13666349 554254 4817588 0 GMP_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 GSH_C0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 GSH_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 GSH_Ellmans_C0 2028973230 7032982415 4327220108 4347412040 6689141561 1910855934 5980351063 515903503 6503241993 7243901097 5270446323 4167054930 2026164371 141480526 842437504 440094624 5622923429 1496419531 4876620475 2924863619 6336408204 2272334227 4607979667 371675879 GSH_Ellmans_C1 434553649 1525340860 914904960 918315220 1447509905 416009383 1311406341 114263365 1407667088 1587432377 1139418288 909023225 434753931 29353312 180246002 96993772 1184779256 312846024 1044781185 630103337 1355640378 496298295 977993418 82629239 GSSG_C0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 GSSG_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 GSSG_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 GTP_C0 121814242 437994498 303227605 89360326 445094235 31050736 352388767 28313928 472238550 342911815 354436514 104442402 34582591 3382669 13838306 13041343 355343303 93203952 316708673 54921792 411261905 94170961 82245962 8119395 GTP_C1 21161264 66640455 47492139 13375664 68068672 4470554 57859052 4767949 73599613 53182138 59949155 16607333 5186579 48438 1998193 1621478 55976514 12088524 49962381 6745619 63283985 12245787 12487235 821139 GTP_C2 6925476 22711762 14067972 3839140 21434899 1372269 19818453 1468837 23380666 17177948 19842510 5317384 1074933 0 429876 191945 17087096 3811552 15645915 1918502 20079505 3449792 3528469 47447 GTP_C3 522249 2337450 1465427 95348 2606364 34145 2343921 0 2836447 2005346 2350771 341412 0 0 39077 0 1623590 87794 1459294 52955 1851041 48535 53542 0 guanine_C0 0 0 0 0 0 0 0 0 0 0 0 244201 0 0 0 0 0 0 0 0 0 0 0 0 guanine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 guanine_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 guanine_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 guanosine_C0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 guanosine_C1 0 0 0 0 0 1036131 0 313270 0 0 0 0 0 0 0 0 0 0 0 0 0 1261453 0 0 guanosine_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 guanosine_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Homocysteine-Ellmans_C0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Homocysteine-Ellmans_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 homocysteine_C0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 homocysteine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 hypoxanthine_C0 16977916 72674428 41832254 44777139 52372930 21410668 70118149 5819993 60752974 129182294 60626342 67935515 30971021 1655225 8315362 3304115 58329436 15445255 48199713 34006064 52934704 28974912 45611817 5505109 hypoxanthine_C1 1961445 5506877 3137846 2984246 5486127 1532029 6113666 643402 4880910 12554183 5733673 6249501 3380520 155419 923076 291957 5250705 1182999 4095587 2213633 4324130 1717293 3948149 400487 hypoxanthine_C2 148981 1063187 205580 386925 747571 185574 2116878 0 1397926 2317559 1364237 1030871 104754 0 32240 0 648439 65852 826619 65222 294949 43823 793619 0 hypoxanthine_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 IMP_C0 6238006 11716416 8069190 8817485 9109089 8259792 13936973 1769388 9645835 9993643 16334135 11323413 15470835 224785 8364198 1373136 7409610 2597404 10184253 6514407 9452576 4549295 16692821 956777 IMP_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4614453 0 IMP_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 IMP_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 inosine_C0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 inosine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 inosine_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 inosine_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 methionine_C0 361770705 942024223 755561213 578003180 1215697657 233047608 1429029599 97900954 2042896116 1739136247 1016499910 568115700 1191966134 45474423 354324687 55279931 2728022926 353487311 1634155436 526723063 2156684909 388470602 2697635580 113267039 methionine_C1 24434292 56605283 46654640 33545483 75899188 13590377 89120755 6198441 119028819 104671406 66299083 35615155 68659684 2831159 21520907 3409781 158969763 20024655 101372929 30709514 121618185 23143384 159058047 6796268 S-Adenosyl-L-Homocysteine_C0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 S-Adenosyl-L-Homocysteine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 S-Adenosyl-L-methionine_C0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 S-Adenosyl-L-methionine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 serine_C0 64506598 151385236 120924380 99239973 189402647 97075892 153088526 32241478 150967626 204686570 142415074 145344280 79702759 15631542 55456295 26481125 123068498 84334643 114802597 139865628 132147480 128112118 111191694 29049964 serine_C1 3703746 7729972 5421175 4575059 11757590 5497486 14702344 1756318 9126467 9287048 8314436 6454495 3467708 104497 2237350 595914 5014881 3308066 6623564 8064514 4503068 5233833 4216496 606413 xanthine_C0 10629537 18417173 14578051 10231789 17376937 2881042 17014676 885446 28388413 18936826 15174550 8863629 5130588 524114 1218764 1347378 17139377 2839188 14825896 4229344 30892559 4714403 17961674 1444461 xanthine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 xanthine_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 xanthine_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 xanthosine_C0 2390858 6426223 4196971 5917346 6743861 1840511 7096337 1473825 7457511 7503668 8966436 5650006 1861678 1527700 2298451 1156662 5054635 1183410 4752403 2698736 6855811 2467797 7234523 826115 xanthosine_C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 xanthosine_C2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 xanthosine_C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name adenine_C0 adenine_C1 adenine_C2 adenine_C3 adenosine_C0 adenosine_C1 adenosine_C2 adenosine_C3 ADP_C0 ADP_C1 ADP_C2 ADP_C3 AICAR_C0 AICAR_C1 AICAR_C2 AMP_C0 AMP_C1 AMP_C2 AMP_C3 ATP_C0 ATP_C1 ATP_C2 ATP_C3 dATP_C0 dATP_C1 dATP_C2 dATP_C3 dimethylglycine_C0 dimethylglycine_C1 dimethylglycine_C2 dTMP_C0 dTMP_C1 dTMP_C2 dTMP_C3 dTTP_C0 dTTP_C1 dTTP_C2 dTTP_C3 formyl_methionine_C0 formyl_methionine_C1 GAR_C0 GAR_C1 GDP_C0 GDP_C1 GDP_C2 GDP_C3 glycine_C0 glycine_C1 GMP_C0 GMP_C1 GMP_C2 GMP_C3 GSH_C0 GSH_C1 GSH_Ellmans_C0 GSH_Ellmans_C1 GSSG_C0 GSSG_C1 GSSG_C2 GTP_C0 GTP_C1 GTP_C2 GTP_C3 guanine_C0 guanine_C1 guanine_C2 guanine_C3 guanosine_C0 guanosine_C1 guanosine_C2 guanosine_C3 Homocysteine-Ellmans_C0 Homocysteine-Ellmans_C1 homocysteine_C0 homocysteine_C1 hypoxanthine_C0 hypoxanthine_C1 hypoxanthine_C2 hypoxanthine_C3 IMP_C0 IMP_C1 IMP_C2 IMP_C3 inosine_C0 inosine_C1 inosine_C2 inosine_C3 methionine_C0 methionine_C1 S-Adenosyl-L-Homocysteine_C0 S-Adenosyl-L-Homocysteine_C1 S-Adenosyl-L-methionine_C0 S-Adenosyl-L-methionine_C1 serine_C0 serine_C1 xanthine_C0 xanthine_C1 xanthine_C2 xanthine_C3 xanthosine_C0 xanthosine_C1 xanthosine_C2 xanthosine_C3 METABOLITES_END #END