#METABOLOMICS WORKBENCH alanyl_20251203_124138 DATATRACK_ID:6823 STUDY_ID:ST004418 ANALYSIS_ID:AN007390 PROJECT_ID:PR002390
VERSION             	1
CREATED_ON             	December 3, 2025, 12:50 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 BCS-treated NALM6, REH or MOLT16 cells with or
ST:STUDY_TITLE                   	without rescue with copper (II) chloride and 15N-amide-glutamine tracing
ST:STUDY_SUMMARY                 	To investigate the effects of bathocuproinedisulfonic acid (BCS) treatment (a
ST:STUDY_SUMMARY                 	copper chelator) on nucleotide synthesis, cells were treated for 6-8 days with
ST:STUDY_SUMMARY                 	vehicle, 50uM BCS, 50uM CuCl2, or 50uM of both. Cells were seeded at 0.125-0.25
ST:STUDY_SUMMARY                 	million/mL and counted and passaged every two to three days. On the day of
ST:STUDY_SUMMARY                 	harvest, cells were seeded at 1.0 million/mL in glutamine-free RPMI-1640 with
ST:STUDY_SUMMARY                 	15N-amide glutamine added back for 4 hours, then subjected to polar
ST:STUDY_SUMMARY                 	metabolomics. This experiment revealed that copper depletion with BCS lead to
ST:STUDY_SUMMARY                 	increased label incorporation and increased abundance of carbamoyl aspartic
ST:STUDY_SUMMARY                 	acid, and reduced label incorporation into purine and pyrimidine nucleotide
ST:STUDY_SUMMARY                 	mono, di and triphosphates.
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                    	8
#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           	-	20250828_QE2_HILIC_NALM6trac_AYW1475	Sample source:NALM6 leukemia cells | treatment:Vehicle	RAW_FILE_NAME(raw file name)=20250828_QE2_HILIC_NALM6trac_AYW1475.raw
SUBJECT_SAMPLE_FACTORS           	-	20250828_QE2_HILIC_NALM6trac_AYW1476	Sample source:NALM6 leukemia cells | treatment:Vehicle	RAW_FILE_NAME(raw file name)=20250828_QE2_HILIC_NALM6trac_AYW1476.raw
SUBJECT_SAMPLE_FACTORS           	-	20250828_QE2_HILIC_NALM6trac_AYW1477	Sample source:NALM6 leukemia cells | treatment:Vehicle	RAW_FILE_NAME(raw file name)=20250828_QE2_HILIC_NALM6trac_AYW1477.raw
SUBJECT_SAMPLE_FACTORS           	-	20250828_QE2_HILIC_NALM6trac_AYW1478	Sample source:NALM6 leukemia cells | treatment:BCS	RAW_FILE_NAME(raw file name)=20250828_QE2_HILIC_NALM6trac_AYW1478.raw
SUBJECT_SAMPLE_FACTORS           	-	20250828_QE2_HILIC_NALM6trac_AYW1479	Sample source:NALM6 leukemia cells | treatment:BCS	RAW_FILE_NAME(raw file name)=20250828_QE2_HILIC_NALM6trac_AYW1479.raw
SUBJECT_SAMPLE_FACTORS           	-	20250828_QE2_HILIC_NALM6trac_AYW1480	Sample source:NALM6 leukemia cells | treatment:BCS	RAW_FILE_NAME(raw file name)=20250828_QE2_HILIC_NALM6trac_AYW1480.raw
SUBJECT_SAMPLE_FACTORS           	-	20250828_QE2_HILIC_NALM6trac_AYW1481	Sample source:NALM6 leukemia cells | treatment:BCS and CuCl2	RAW_FILE_NAME(raw file name)=20250828_QE2_HILIC_NALM6trac_AYW1481.raw
SUBJECT_SAMPLE_FACTORS           	-	20250828_QE2_HILIC_NALM6trac_AYW1482	Sample source:NALM6 leukemia cells | treatment:BCS and CuCl2	RAW_FILE_NAME(raw file name)=20250828_QE2_HILIC_NALM6trac_AYW1482.raw
SUBJECT_SAMPLE_FACTORS           	-	20250828_QE2_HILIC_NALM6trac_AYW1483	Sample source:NALM6 leukemia cells | treatment:BCS and CuCl2	RAW_FILE_NAME(raw file name)=20250828_QE2_HILIC_NALM6trac_AYW1483.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1499	Sample source:MOLT16 leukemia cells | treatment:Vehicle	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1499.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1500	Sample source:MOLT16 leukemia cells | treatment:Vehicle	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1500.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1501	Sample source:MOLT16 leukemia cells | treatment:Vehicle	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1501.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1502	Sample source:MOLT16 leukemia cells | treatment:BCS	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1502.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1503	Sample source:MOLT16 leukemia cells | treatment:BCS	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1503.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1504	Sample source:MOLT16 leukemia cells | treatment:BCS	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1504.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1505	Sample source:MOLT16 leukemia cells | treatment:BCS and CuCl2	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1505.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1506	Sample source:MOLT16 leukemia cells | treatment:BCS and CuCl2	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1506.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1507	Sample source:MOLT16 leukemia cells | treatment:BCS and CuCl2	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1507.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1523	Sample source:REH leukemia cells | treatment:Vehicle	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1523.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1524	Sample source:REH leukemia cells | treatment:Vehicle	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1524.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1525	Sample source:REH leukemia cells | treatment:Vehicle	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1525.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1526	Sample source:REH leukemia cells | treatment:BCS	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1526.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1527	Sample source:REH leukemia cells | treatment:BCS	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1527.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1528	Sample source:REH leukemia cells | treatment:BCS	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1528.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1529	Sample source:REH leukemia cells | treatment:BCS and CuCl2	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1529.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1530	Sample source:REH leukemia cells | treatment:BCS and CuCl2	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1530.raw
SUBJECT_SAMPLE_FACTORS           	-	20250829_QE2_HILIC_REHMOLTtrac_AYW1531	Sample source:REH leukemia cells | treatment:BCS and CuCl2	RAW_FILE_NAME(raw file name)=20250829_QE2_HILIC_REHMOLTtrac_AYW1531.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	1-1.5 million cells from culture were collected via centrifugation for 20
CO:COLLECTION_SUMMARY            	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 for the first 12 days. On day 12, cells were seeded in RPMI-1640 with 10%
CO:COLLECTION_SUMMARY            	dialyzed FBS and penicillin/streptomycin with the noted treatments. Glutamine
CO:COLLECTION_SUMMARY            	tracing was performed for 4 hours on day 14.
CO:SAMPLE_TYPE                   	Leukemia cells
#TREATMENT
TR:TREATMENT_SUMMARY             	Culture of NALM6, REH or MOLT16 cells for 6-8 days in RPMI-1640 media with 10%
TR:TREATMENT_SUMMARY             	FBS and penicillin/streptomycin. On the day of harvest, media was changed to
TR:TREATMENT_SUMMARY             	RPMI-1640 with 10% dialyzed FBS and labelled 15N-amide-glutamine at RPMI-1640
TR:TREATMENT_SUMMARY             	levels (2mM). Amino acid tracing was performed for 4 hours.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Cell pellet was resuspended in 400uL of LC-MS grade methanol (supplemented with
SP:SAMPLEPREP_SUMMARY            	isotopically-labelled amino acid standards [Cambridge Isotope Laboratories,
SP:SAMPLEPREP_SUMMARY            	MSK-A2-1.2], aminopterin, reduced glutathione standard [Cambridge Isotope
SP:SAMPLEPREP_SUMMARY            	Laboratories, CNLM-6245-10]), and then 100uL of LC-MS grade water with 25 mM
SP:SAMPLEPREP_SUMMARY            	Ammonium Acetate and 2.5 mM sodium ascorbate was added. The sample was vortexed
SP:SAMPLEPREP_SUMMARY            	for 10 seconds. Samples were then centrifuged for 10 minutes at 18,000 g to
SP:SAMPLEPREP_SUMMARY            	pellet cell debris. The supernatant was transferred to a new tube and dried on
SP:SAMPLEPREP_SUMMARY            	ice using a liquid nitrogen dryer. The dried metabolites were then resuspended
SP:SAMPLEPREP_SUMMARY            	in 30uL and 2uL was 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 × 10e6, 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	Peak area
MS_METABOLITE_DATA_START
Samples	20250828_QE2_HILIC_NALM6trac_AYW1475	20250828_QE2_HILIC_NALM6trac_AYW1476	20250828_QE2_HILIC_NALM6trac_AYW1477	20250828_QE2_HILIC_NALM6trac_AYW1478	20250828_QE2_HILIC_NALM6trac_AYW1479	20250828_QE2_HILIC_NALM6trac_AYW1480	20250828_QE2_HILIC_NALM6trac_AYW1481	20250828_QE2_HILIC_NALM6trac_AYW1482	20250828_QE2_HILIC_NALM6trac_AYW1483	20250829_QE2_HILIC_REHMOLTtrac_AYW1499	20250829_QE2_HILIC_REHMOLTtrac_AYW1500	20250829_QE2_HILIC_REHMOLTtrac_AYW1501	20250829_QE2_HILIC_REHMOLTtrac_AYW1502	20250829_QE2_HILIC_REHMOLTtrac_AYW1503	20250829_QE2_HILIC_REHMOLTtrac_AYW1504	20250829_QE2_HILIC_REHMOLTtrac_AYW1505	20250829_QE2_HILIC_REHMOLTtrac_AYW1506	20250829_QE2_HILIC_REHMOLTtrac_AYW1507	20250829_QE2_HILIC_REHMOLTtrac_AYW1523	20250829_QE2_HILIC_REHMOLTtrac_AYW1524	20250829_QE2_HILIC_REHMOLTtrac_AYW1525	20250829_QE2_HILIC_REHMOLTtrac_AYW1526	20250829_QE2_HILIC_REHMOLTtrac_AYW1527	20250829_QE2_HILIC_REHMOLTtrac_AYW1528	20250829_QE2_HILIC_REHMOLTtrac_AYW1529	20250829_QE2_HILIC_REHMOLTtrac_AYW1530	20250829_QE2_HILIC_REHMOLTtrac_AYW1531
Factors	Sample source:NALM6 leukemia cells | treatment:Vehicle	Sample source:NALM6 leukemia cells | treatment:Vehicle	Sample source:NALM6 leukemia cells | treatment:Vehicle	Sample source:NALM6 leukemia cells | treatment:BCS	Sample source:NALM6 leukemia cells | treatment:BCS	Sample source:NALM6 leukemia cells | treatment:BCS	Sample source:NALM6 leukemia cells | treatment:BCS and CuCl2	Sample source:NALM6 leukemia cells | treatment:BCS and CuCl2	Sample source:NALM6 leukemia cells | treatment:BCS and CuCl2	Sample source:MOLT16 leukemia cells | treatment:Vehicle	Sample source:MOLT16 leukemia cells | treatment:Vehicle	Sample source:MOLT16 leukemia cells | treatment:Vehicle	Sample source:MOLT16 leukemia cells | treatment:BCS	Sample source:MOLT16 leukemia cells | treatment:BCS	Sample source:MOLT16 leukemia cells | treatment:BCS	Sample source:MOLT16 leukemia cells | treatment:BCS and CuCl2	Sample source:MOLT16 leukemia cells | treatment:BCS and CuCl2	Sample source:MOLT16 leukemia cells | treatment:BCS and CuCl2	Sample source:REH leukemia cells | treatment:Vehicle	Sample source:REH leukemia cells | treatment:Vehicle	Sample source:REH leukemia cells | treatment:Vehicle	Sample source:REH leukemia cells | treatment:BCS	Sample source:REH leukemia cells | treatment:BCS	Sample source:REH leukemia cells | treatment:BCS	Sample source:REH leukemia cells | treatment:BCS and CuCl2	Sample source:REH leukemia cells | treatment:BCS and CuCl2	Sample source:REH leukemia cells | treatment:BCS and CuCl2
ADP_N0	13685148	8555446	5107292	48960282	27423751	47305889	23000954	23208705	5612506	23813714	5307070	23461952	20810609	42723411	25002331	9565358	26491737	9738671	29469364	30338868	29824644	31287952	31407632	32655529	20929031	19547161	19832502
ADP_N1	0	0	0	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_N2	12401524	7886194	4534765	7202801	4340605	6946147	23194593	24223066	4681957	1034437	182816	998660	575660	1548090	607022	212668	534578	148886	24045637	24799737	25282565	11601748	11710423	11749715	14976996	14898229	14720815
ADP_N3	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
AICAR_N0	39456	6980	115559	644158	400434	691111	28625	15677	0	0	0	0	0	13698	54031	0	12597	11853	26192	0	0	779531	658286	637953	0	0	0
AICAR_N1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
AICAR_N2	211134	128026	112640	2980519	1188419	3026824	3838019	3758034	979868	118879	12660	171703	35594	62675	51515	18286	37065	25198	231296	233759	108551	3629661	4104333	4146203	2558720	2013266	1936878
AICAR_N3	0	0	0	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_N0	4526647	3898631	3920624	11286533	2319407	5037786	5811521	1840404	3259610	3824029	672607	3588137	2468944	7996733	2368705	880545	4301606	2775700	1872259	2259034	3674042	4746760	3494839	2545772	1912046	3280114	2448754
AMP_N1	0	0	0	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_N2	3963653	3803228	3414924	1525640	648748	711384	6223906	2135978	3332626	863061	695560	1476952	1348191	992941	726451	673575	809267	458974	1358603	1492073	2537126	1796271	1166053	1117682	1365437	2287679	1500009
AMP_N3	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
ATP_N0	37759419	186294754	107049395	153381190	223876750	143507064	163399425	187481487	78535643	247222715	254186866	204714378	202203520	254720848	220536886	271767726	266257845	52954600	212220370	210027510	209801185	144345961	143358026	141704845	139041102	143255731	132115157
ATP_N1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
ATP_N2	36650364	176983716	102050444	23701340	35914723	22943516	169862740	194728256	80279810	13670974	14832821	11238446	10850317	14193405	12407541	9748459	9643036	1403276	171280934	166182676	167781787	53644866	53108716	54038153	99751728	103646541	98414912
ATP_N3	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
CDP_N0	343329	956849	925325	836257	972172	852491	753871	788660	762692	983265	1016100	992327	1860111	2288643	2052759	959257	1005603	681084	504219	425568	548751	549846	550954	589463	437039	412859	395697
CDP_N1	206169	918257	888632	421690	405131	376121	659863	777291	668937	35185	33779	0	516601	584622	491422	61713	143379	98394	368058	379209	425426	252538	253237	240484	314563	257298	295676
CDP_N2	527812	1646082	1471777	34989	63285	30749	1338615	1412989	1206597	0	0	0	26617	0	0	0	9452	0	1125674	1062799	989370	159755	139070	142569	179436	301717	252579
CDP_N3	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
CTP_N0	11728214	15544097	13303272	5141073	7337259	4685721	10851545	12778088	11524869	16594455	16798853	13713442	22418316	25213425	21475131	19431027	19226449	17912062	9922058	10282259	9575800	8602920	8947710	8874054	6972053	7180854	6415541
CTP_N1	9650018	13040133	10763772	1982445	2895557	1890994	9384872	10469366	9426366	2975354	3172066	2562515	4788655	4947478	4496497	3096922	2937366	2935522	9126241	9196146	8630587	5480306	5592821	5325911	6371329	6310421	5831378
CTP_N2	16343739	21619169	18776065	976590	1404902	900622	16556450	18349947	16053162	0	0	0	0	0	0	0	0	0	17934711	17496731	17724488	8378382	9067284	8315951	11768981	11646844	10499524
dihydroorotate_N0	0	34990	33398	5847256	6206373	5848305	103722	0	0	0	83430	0	8711408	9568149	7238602	0	76743	41357739	41413	101773	97693	0	0	0	130951	0	0
dihydroorotate_N1	0	0	0	8275666	8669509	9108192	0	0	0	0	0	0	2090724	2681973	1825100	0	0	140488	19071	0	0	0	0	0	0	0	0
dihydroorotate_N2	0	0	0	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_N0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	3704190	0	0
dTMP_N1	1826433	0	0	2034168	0	0	0	0	0	0	0	0	1437583	0	0	0	0	1549191	0	0	0	0	0	0	2386343	0	0
dTMP_N2	3584559	0	0	6113167	3120754	0	0	0	0	0	0	0	5114369	0	0	6073931	0	5606046	0	0	0	0	0	0	1790972	0	0
dUMP_N0	309290	0	0	0	0	15822	0	0	0	0	0	0	7703	0	0	0	0	0	0	0	0	0	0	0	0	0	219991
dUMP_N1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
dUMP_N2	0	0	0	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_N0	0	0	0	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_N1	0	0	0	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_N2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
GDP_N0	1995127	2757232	2523105	6527905	7185460	6595040	2202798	2165415	1993538	3100559	3237410	3138673	5438871	5766945	5375145	3686666	3773693	3616884	2441125	2480519	2465962	2772911	3293930	2867393	2234303	2109544	1918408
GDP_N1	1935287	4239326	3764626	4692522	5254034	3573248	3881214	3948868	3620231	3757812	3722058	3252890	3571454	4130450	3267274	3848818	3615734	3522746	4555107	4153983	4096154	4815450	5099048	4762212	4011624	4084333	3781423
GDP_N2	433980	525392	489028	42909	57038	34879	400823	412039	351474	0	9243	5980	0	9849	0	8764	0	13096	238000	200869	284267	83259	130613	76010	115181	123085	114411
GDP_N3	3257081	4371537	3891710	1242849	1165944	1292493	3531768	3423636	3246313	27475	62873	62443	134049	147551	163721	36118	43388	32236	2459486	2441097	2403849	1352259	1465109	1470984	1566073	1548747	1272707
GDP_N4	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
glutamine0	146534478	996012583	121598917	195239522	96836730	74844072	228792702	133596970	112604414	196928698	60487647	195059272	60097942	220799176	80985562	81459364	229693909	163136319	141173177	102830838	164727762	193489439	163068881	148303085	116506071	167931970	168606183
glutamine1	4299981966	3806478273	3630711445	6628094377	3412249696	2604992232	7143335728	4136736754	3558938718	8324489760	2507826272	8496053033	2615626757	9310954630	3523408940	3223256301	9343644180	6561831075	5923280581	4451582199	7367348422	7839861770	6732515459	6127555245	4616934589	6817646104	6518934756
glutamine2	17332346	15803758	15085376	28649840	14099727	10982333	29150416	16075203	14590569	38531885	9627556	37278721	12316086	36790134	13930716	12605357	38889495	25677034	27396101	18288414	31498209	35543544	29576650	26433548	20713084	30231537	29442912
GMP_N0	132692	559544	369631	3266541	3235061	3194596	777417	455001	412427	571896	309312	508480	1263713	1693955	1689063	738390	709556	138242	604477	582095	528743	976188	989394	807583	876168	906751	654848
GMP_N1	0	0	0	210604	185232	202723	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	27871
GMP_N2	325791	766506	666694	869950	603756	939764	842562	892515	490332	884000	813447	763336	720595	607415	555731	826375	773784	157509	938557	996430	1093878	923602	1235530	843926	1174938	1001499	814458
GMP_N3	12085	294644	135573	43840	0	28292	169749	274917	23098	0	17540	23216	0	33917	0	17965	0	31886	0	0	42639	0	50465	0	12858	0	32358
GMP_N4	0	0	0	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_N0	15332291	16820641	13482827	22505975	35254545	20456214	10894478	12242646	11058138	21268204	20758839	16602296	21963331	25127932	20436003	24090488	22832483	21985125	11163687	10489135	10852578	13094573	13696453	12838543	8976738	9478914	8230913
GTP_N1	104596	168791	594969	5631596	8621742	4873340	499681	96431	319226	1142188	1343605	1261340	2348629	2591354	2663901	721388	446116	415350	2472965	2388101	2342348	5273150	5381723	5287132	2873756	3005661	2533902
GTP_N2	3568920	3895616	3318461	86548	16825	34300	2524803	2701899	2392092	0	44576	22069	0	0	9810	14421	46118	11606	2057777	2061300	2313755	1238679	1401506	1175087	1210858	1245339	1142269
GTP_N3	28603681	30710532	25202803	4685785	7297752	4383358	20790164	23150256	20698639	761938	705776	592658	699446	775013	624966	333590	377995	327363	17951106	15860512	15986398	8963431	9177102	9390441	9352492	9827681	8513336
GTP_N4	0	0	0	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_N0	23858	51933	44829	41513564	28509610	45151351	24097	31819	121392	73211	0	0	2061988	4556185	3571790	330723	417351	56062	53961	135905	434842	17888399	19034360	17355116	549243	670482	518686
IMP_N1	0	0	0	4034541	2920388	4212204	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	2722976	2349182	2547583	0	0	0
IMP_N2	1471729	784122	769819	73020754	51048566	80202336	1969517	2154460	406001	0	0	0	0	0	57751	0	0	0	0	1882541	1268448	53304168	54389181	50433798	0	0	0
IMP_N3	0	0	0	0	0	0	0	0	0	0	0	152321	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
orotate_N0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
orotate_N1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
orotate_N2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
UDP_N0	458092	2106281	1165616	2122774	2265578	2093363	1909655	1963996	739564	1901190	1715210	2020801	6401198	7407860	6389942	2197770	2577009	307956	1103149	1015771	1015856	1171031	1223258	1301250	893891	946730	801461
UDP_N1	1326266	5761381	3673858	2020720	2389374	1939033	5641567	5720633	2544237	162835	146124	162412	67757	137233	89314	79405	59862	18016	3618046	3634342	3630074	2842741	2821078	2843116	2063753	2347934	2257364
UDP_N2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
UMP_N0	335912	227110	159438	595056	336792	469663	413743	581043	134801	273964	92606	286528	968608	1846871	997614	162039	462226	235462	107957	113608	45867	69185	170477	193839	128843	174632	200024
UMP_N1	1046364	721988	564606	574099	247787	556164	1734676	1683263	509189	15499	8870	17981	34358	52424	0	0	23119	0	555070	494102	514591	540910	557481	598018	424377	488363	525558
UMP_N2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Ureidosuccinic_acid_N0	0	0	0	51301670	50394876	52740250	0	0	0	0	0	0	62039698	79452478	68429089	0	0	0	0	0	0	0	0	0	0	0	0
Ureidosuccinic_acid_N1	541150	2391869	1641048	102440271	105750310	105363591	1203818	1255888	455135	0	0	0	18178639	22978531	20950099	0	0	0	1333839	1214263	948801	573201	702783	422538	180353	409039	327381
Ureidosuccinic_acid_N2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
UTP_N0	36293541	42590335	35523158	11464127	17582200	11729770	30861849	34283731	30832952	39232685	41313340	34614528	70444642	79874850	64449300	45809714	45349084	42489932	26989235	27196657	26475405	22551844	21603435	21850858	19264622	20722711	19025904
UTP_N1	124333968	143498470	117966806	12567541	18519341	11662389	102304583	112967842	103357974	10182384	10773083	9085509	310418	0	0	9200119	8875852	8507936	91400097	90456651	89397630	56054764	55108297	56458027	55968074	59730964	53570119
UTP_N2	0	0	0	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
ADP_N0
ADP_N1
ADP_N2
ADP_N3
AICAR_N0
AICAR_N1
AICAR_N2
AICAR_N3
AMP_N0
AMP_N1
AMP_N2
AMP_N3
ATP_N0
ATP_N1
ATP_N2
ATP_N3
CDP_N0
CDP_N1
CDP_N2
CDP_N3
CTP_N0
CTP_N1
CTP_N2
dihydroorotate_N0
dihydroorotate_N1
dihydroorotate_N2
dTMP_N0
dTMP_N1
dTMP_N2
dUMP_N0
dUMP_N1
dUMP_N2
GAR_N0
GAR_N1
GAR_N2
GDP_N0
GDP_N1
GDP_N2
GDP_N3
GDP_N4
glutamine0
glutamine1
glutamine2
GMP_N0
GMP_N1
GMP_N2
GMP_N3
GMP_N4
GTP_N0
GTP_N1
GTP_N2
GTP_N3
GTP_N4
IMP_N0
IMP_N1
IMP_N2
IMP_N3
orotate_N0
orotate_N1
orotate_N2
UDP_N0
UDP_N1
UDP_N2
UMP_N0
UMP_N1
UMP_N2
Ureidosuccinic_acid_N0
Ureidosuccinic_acid_N1
Ureidosuccinic_acid_N2
UTP_N0
UTP_N1
UTP_N2
METABOLITES_END
#END