#METABOLOMICS WORKBENCH alanyl_20250326_081441 DATATRACK_ID:5782 STUDY_ID:ST003823 ANALYSIS_ID:AN007475 PROJECT_ID:PR002390
VERSION             	1
CREATED_ON             	December 17, 2025, 1:25 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 SEM cells with or without genetic
ST:STUDY_TITLE                   	overexpression of AOX.
ST:STUDY_SUMMARY                 	To investigate whether the effects of bathocuproinedisulfonic acid (BCS)
ST:STUDY_SUMMARY                 	treatment (a copper chelator) on steady state metabolite levels can be rescued
ST:STUDY_SUMMARY                 	by overexpression of alternative oxidase (AOX), SEM cells were engineered to
ST:STUDY_SUMMARY                 	overexpress red fluorescent protein (RFP) or AOX and treated for 14 days with
ST:STUDY_SUMMARY                 	vehicle or 50uM BCS. Cells were seeded at 0.25 million/mL and counted and
ST:STUDY_SUMMARY                 	passaged every two days. This experiment revealed that AOX overexpression
ST:STUDY_SUMMARY                 	significantly rescues levels of nucleotides and reduces ureidosuccinic
ST:STUDY_SUMMARY                 	acid/carbamoyl aspartic acid to near normal.
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           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1184	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:RFP	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1184.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1185	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:RFP	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1185.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1186	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:RFP	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1186.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1187	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:RFP	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1187.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1188	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:RFP	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1188.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1189	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:RFP	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1189.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1190	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:AOX	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1190.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1191	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:AOX	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1191.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1192	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:AOX	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1192.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1193	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:AOX	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1193.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1194	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:AOX	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1194.raw
SUBJECT_SAMPLE_FACTORS           	-	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1195	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:AOX	RAW_FILE_NAME(raw file name)=20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1195.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	One million cells from culture were collected via centrifugation for 20 seconds
CO:COLLECTION_SUMMARY            	at 18,000xG at 4C, washed once with ice-cold 0.9% NaCl, and collected via
CO:COLLECTION_SUMMARY            	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 treated for 14 days with vehicle or 50uM BCS. Culture media was
TR:TREATMENT_SUMMARY             	RPMI-1640 containing 10% FBS and penicillin and streptomycin in 6-well plates.
TR:TREATMENT_SUMMARY             	Cells were seeded at 0.25 million/mL in 4mL and counted and passaged every two
TR:TREATMENT_SUMMARY             	days.
#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 × 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	Normalized peak area
MS_METABOLITE_DATA_START
Samples	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1184	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1185	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1186	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1187	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1188	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1189	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1190	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1191	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1192	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1193	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1194	20240812_QE2_HILIC_BCSPU_BCSAOX_AYW1195
Factors	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:RFP	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:RFP	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:RFP	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:RFP	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:RFP	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:RFP	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:AOX	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:AOX	Sample source:SEM leukemia cells | guideRNA:Vehicle | overexpression:AOX	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:AOX	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:AOX	Sample source:SEM leukemia cells | guideRNA:BCS | overexpression:AOX
2-hydroxyglutarate	671482329.6	796955752.4	694338281.7	649538327.7	536607282.7	1091380063	823483162.5	687710823.4	733998465.7	748440566.2	873940629.5	740374282.9
3-methyl.2.oxopentanoic_acid	18806483.13	21862165.36	22451898.41	8170666.609	9129973.322	14345727.35	20700803.33	22444938.02	22339265.78	17302400.81	16352612.87	22475004.08
3-phosphoglycerate	2726862.937	2466027.607	2488685.37	1512532.16	1923090.619	1453894.569	3008952.664	3065600.689	2902215.241	4989382.831	5768051.442	5245318.188
Acetyl.CoA	4200463.931	3948707.384	4081641.672	2168950.102	2784317.544	2728621.027	4333669.222	4236577.611	4397147.572	3393720.371	3022872.794	2896923.322
acetylcarnitine	71563470.68	78901770.35	69734673.26	66640738.96	73798453.59	84972691.35	67330883.37	68435756.52	83062457.84	74285906.23	68923587.04	78467625.35
ADP	52244420.81	54853094.73	52448450.41	68482987.67	73172185.22	71494014.73	43792427.83	45908368.53	43053536.17	42457700.23	44399074.78	40527224.28
alanine	93421511.66	93556553.52	90160576.27	87363027.13	76047247.75	85605386.35	97197120.3	95276652.65	83139611.09	81938827.05	81917204.69	73804582.46
Alpha.Ketoglutarate	33571366.12	23527679.2	32131454.21	10011532.26	12066941.51	12536316.28	23541086.22	32055205.05	37177520.8	25955384.56	23436980.66	24375256.28
AMP	1424583.522	1676218.731	1614809.317	4884753.179	4891257.035	5317738.061	693816.5168	1150607.025	1557342.41	1423617.516	1298317.462	1922569.271
arginine	834290350.9	920715879.5	876322439	699276187.9	703087765	702548099.4	997292921.4	895721514.5	868235047.1	814357707.7	796297652.2	840102612.8
argininosuccinate	12858787.6	10742350.08	12509475.12	9402441.075	8618387.439	11426124.4	13562516.32	12832502.14	12400776.3	10570047.76	11017720.98	11076140.76
ascorbate_unc	1.01964E+11	94443685741	1.03053E+11	94363901331	73045900647	83654254143	1.04862E+11	1.02061E+11	82714960797	91481042004	1.06988E+11	85203586324
asparagine	1315076235	1211448861	1410896753	958723276.4	945399163.5	975450169.4	1320533485	1351709053	1271016062	1047980277	1081072188	1010915212
aspartate	29430973.47	30312802.47	28295947.75	9393935.303	9133780.63	8135728.148	29190441.64	28142180.99	28696794.06	18160720.34	18033320.83	17512096.93
asymmetric.dimethylarginine	8608346.704	8658585.685	9003617.01	6198983.719	6548545.635	6466799.797	8419268.894	8706119.148	8993912.173	7333244.951	7020360.555	7687265.964
ATP	238375614.8	221254234.9	254067185.4	174676286.9	196854148.3	111829379.3	192568061.9	268874284.4	215589808.8	202536252.3	198660109.1	180269565.4
beta.alanine	24718735	23833869.24	25788765.96	25252543.03	27335487.56	30235632.49	27171058.81	28266114.3	29188581.2	31630838.51	28658916.84	28538859.7
betaine	860399835.3	850177042.9	845435921.1	499432495.7	515803212.3	520562642.9	829084418.5	824788963.2	884084643.3	712825379.1	651287028	662440931.2
Butyrylcarnitine	74710234	70232375.23	73868414.28	48663435.03	52740281.44	50841410.98	76375501.15	78065782.96	77122122.23	61783479.94	60871412.49	57743451.37
C5.Carnitines	13427475.36	12445750.91	11859972.68	11993406.32	10905404.05	12483798.53	11025405.15	11345433.11	10485669.32	5257913.088	4959975.296	5003060.741
carnitine	147594977.9	144287397	138617729.9	153848859.7	163392215	143285395.6	141883357	145079607.8	137749119.1	154596492	145383809.8	146939990.8
carnosine	2720127.409	3526466.258	3620297.054	3294764.391	3287773.112	2558176.093	2684448.241	2804924.2	3518172.773	2716839.065	2945637.47	3988507.802
CDP.choline	326411.9436	321558.3851	253298.0691	1068178.208	1032467.757	852576.2698	407947.0082	276018.6442	186618.426	936504.657	959947.065	943219.1366
CDP	1631857.783	1785258.521	1544309.898	1355143.504	1178062.164	1154574.388	1313663.562	1313572.143	1277379.695	1177532.966	1123659.222	1070324.182
cis.aconitate	5711288.956	5440757.421	6028224.231	2002507.774	1817720.127	833595.5468	5178336.544	6184311.58	5343101.452	4510045.29	4070217.061	4395805.027
citrate	254758085.2	238817432.1	267358494.3	65881323.89	64807055.78	63506386.94	229820968.7	275482089.2	262740516.1	220664330.6	207460074.4	206104632.3
citrulline	15471432.16	16579851.51	14727071.41	15535607.28	12279145.13	17235098.33	16536098.65	13452247.07	13044496.31	15424787.9	16436977.9	15495149.41
CMP	216245.4034	252274.0754	198821.8059	409785.2764	300298.0788	352876.6003	236540.495	153821.8262	191233.4299	197821.2209	164942.677	138105.003
creatine	208747584.4	209267920.4	257637351.2	245550695.3	183365164.8	182282395.1	199996618.9	216677489.9	225548490.2	220851092.7	203980194.4	258393178.5
creatinine	40434516.09	45128883.11	41963331.31	34834013.07	33899283.76	37204397.19	41796237.54	41957463.59	40925184.51	40760316.68	39666578.06	41062140.86
CTP	22463534.55	20103576.21	23392001.87	7709985.696	8048176.832	5385638.673	19342327.16	24039917.63	20516757.88	18731747	17868821.94	16168837.13
Cys_Ell	11643938.59	19335244.96	15485875.99	15592942.19	15138874.41	18233423.27	15442925.18	12971576.88	16693941.96	18474458.64	18180402.05	28012694.85
cystathionine	17435868.65	15327969.29	16532022.6	7713384.299	6383463.004	7390305.064	20121124.21	18065626.21	18772762.55	11566260.01	12313170.17	11492265.62
cytidine	72868.42902	82467.18019	81755.16288	250544.418	111935.031	359963.9665	354922.9744	39290.74112	84378.78389	387115.271	253108.9543	118341.0802
dihydroxyacetone_phosphate	3494217.057	3440146.427	3696511.441	7017449.205	9328649.53	6936638.691	3514854.709	4334343.617	3873503.917	9162925.314	7661987.391	8322701.8
dTMP	2532260.354	2816897.943	3223749.243	3071018.428	2307878.569	2871138.417	3053045.011	3264469.088	2496702.112	2898309.922	3367646.687	3090880.386
F6P_or_G1P	2661645.31	2807666.708	2339783.667	7717099.131	7918651.896	6362845.61	2879194.062	2626242.784	2427984.211	6039752.474	5341771.965	4998674.472
FAD	1506911.799	1582268.117	1631488.473	1662973.024	1719158.687	1353464.468	1742300.711	1793141.294	1664477.658	1678129.135	1750887.962	1463767.068
Fructose.1.6.bisphosphate	1096899.034	1005780.843	906352.2863	899150.4398	811452.7305	894370.7501	1200350.02	1104818.108	1097987.953	1469707.978	1363826.279	1315965.007
fumarate	34614376.16	29282496.57	36508826.3	23752389.01	23123574.47	23415129.55	39405166.01	39820414.29	33760859.71	29744269.72	26845210.56	29680658.52
gamma.aminobutyric_acid	69076514.13	74286100.7	69374730.39	55626652.55	66732867.18	62915590.17	68174379.04	65785664.93	67090746.15	76041394.66	72938844.21	67667760.74
GDP	3363089.97	3580286.58	3491538.572	6176664.608	5908877.891	5670048.64	2643418.224	2645947.543	2552407.612	2941017.589	2864864.402	2957206.96
Glucose.6.phosphate	296420.5103	294471.9403	294727.2572	438781.1241	459588.8362	347753.5919	274208.4883	279515.3776	298499.7807	334400.665	319215.3812	288336.5319
glutamate	848943720.8	862093877.4	799993518.8	614397584.3	609389336.2	557028825.5	868936990.4	848478272.7	857732801.4	733127032.9	705183508.3	670798794.2
glutamine	3273559714	3340395849	3411213487	2447504371	2571742942	2462914824	3368589058	3343904191	3177136687	2744825572	2701875350	2608293138
glyceraldehyde_3.phosphate	3494955.491	3402507.458	3696721.46	6951801.966	9273088.799	6937355.727	3515505.9	4335019.446	3874122.623	9164290.836	7662890.11	8323581.012
glycerate	27121721.52	30483066.48	26533517.85	26747595.67	21201252.07	27079176.86	31592166.57	26678461.92	23862797.8	29402723.72	32787591.17	23819794.54
Glycerol.3.Phosphate	11508941.29	10158744.75	10622153.53	57154496.96	58314951.13	64831116.91	8883150.566	8500093.372	8937339.563	20664438.89	20540053.39	23002888.18
Glycerophosphocholine	1461852696	1563861343	1347514452	1003905879	986346150.1	1004965300	1648004628	1482887140	1357172670	1360037451	1325085496	1244875002
glycine	11331664.44	10587399.92	9726027.902	8601107.605	7461312.59	7057216.141	12323618.94	9876321.656	9222350.836	10783230.29	10516763.04	9614668.457
GMP	497931.8751	475502.0151	523598.9684	2178280.621	2805056.792	2501801.097	387345.667	477144.9549	439464.9743	479605.2058	655245.7721	476999.9517
GSH_Ellmans	304872417.6	292741580.9	286707159.3	267923624.7	286710739.3	239815351.9	304898080.9	320986816.7	280415097.6	297749558.8	295067212.7	260681306.9
GSSG	4098563.556	3644315.759	3437493.464	3675985.885	4290908.059	2217068.348	3414199.94	4205696.134	5370242.919	3391194.97	3517228.032	2503586.969
GTP	18179818.71	15325395.67	18986431.75	15624453.97	18004183.93	7271635.485	12514605.61	19354619.08	14055316.69	15127405.4	14385319.75	13724232.13
histidine	317432608.8	360945623.7	297497670.9	265222355.4	232806830	281220775.1	338088911	292312363.7	327855096.5	307469432.4	299622302.6	306287092.8
hypotaurine	33873721.58	41718760.44	40968432.87	34799377.21	26882549.5	93468966.77	34152375.3	29184342.96	87300598.81	42373096.59	43320030.75	108924361.9
IMP	601893.4149	569921.976	633096.3067	2830180.993	2838302.716	2490919.892	620488.3713	751219.2537	789686.608	2114786.212	1889661.91	1988031.449
isoleucine	656789941	665291978.9	621896778.4	560660062.4	523671078.2	741279281.1	728198391.6	636338318.3	630087869	646301201.4	583929921.1	587199432
lactic_acid	338490673.1	338916861.2	353579356.8	337235191.4	346659451.4	523952677.3	336013874.7	339165529.6	428920252.9	302708406.2	513539222.3	650525249.9
lysine	13105255.04	15749711.04	14380260.86	12090271.13	12995624.11	11545338.1	16135975.81	14298292.65	13205080.73	13802428.66	13042451.4	15459252.29
mannose	4676718.333	6113705.537	5217419.129	4729077.726	3474217.39	4057902.22	4892940.629	4869982.841	4586374.229	3844090.339	4411169.533	4744806.026
Methionine.sulfoxide	75897776.46	83524550.29	78874403.09	50893576.2	55019951.96	73994220.36	81027603.16	71141040.46	72542311.12	71546824.93	64829328.18	62299614.78
methionine	237521513.7	242471048	228165511.2	189744597.8	184995452.8	267796026	251594812.8	225224901.9	244151104.9	221966454.7	199530895.1	237314812.7
methylthioadenosine	7635202.36	8061141.283	6362858.661	4492456.69	4037506.909	5019616.784	8378624.904	6611328.771	6648402.322	7420018.188	6635212.735	5366843.048
myo.inositol	232347112.1	214881510.9	217095760.7	234463320	229813301.7	220013946.8	220029024.4	222172184.8	220065627.6	192001722.3	190644376.1	193686540.8
N.acetylglutamate	6782214.931	7086688.805	6684016.098	7360950.149	5693516.922	6464150.681	7711972.213	6988158.81	7658698.304	9456511.546	10660841.08	9117656.003
NAD.	98133092.94	89612687.53	97498191.4	75632985.86	79239021.04	72837247.01	92606279.63	98175393.58	95170754.68	97393569.4	90916136.03	88894223.69
NADH	16241323	13050391.23	15282543.73	24240963.01	25597499.5	20963692.78	13376323.55	15716518.56	14507587.06	11843079.15	13492738.64	14016745.53
NADP	699811.2625	724121.4113	610997.4452	718030.6435	658831.4309	684170.6256	753576.9326	677235.5483	671534.2303	622814.2341	672944.5881	553886.9995
NADPH	4100406.058	3691500.715	3930954.814	2729229.526	3011536.116	2550254.106	3750768.679	4293891.257	4064896.048	3080040.828	2998833.519	3060952.86
niacinamide	33645468.85	34299521.41	36393659.69	35225504.51	35539285.65	40616159.41	39649877.24	40831452.48	40792234.9	39342815.19	40713861.8	48238363.69
O.phosphoethanolamine	792355.7445	1059763.546	623882.6824	18818898.2	22422061.53	15232607.26	807791.1183	716067.8182	477556.8684	3384923.36	3362329.982	2736056.745
ornithine	39215124.16	38365511.07	40080644.96	29798838.72	30709536.83	29752922.68	39821895.48	37195815.35	37241016.49	30982179.45	30393403.32	35771919.13
palmitate	119354960.7	87351641.16	177673275.8	106214953.4	54364771.34	85453455.26	92319021.4	98521316.02	109204385.6	94227128.53	243286156.9	121582461.5
Pantothenate	388350426	392310012.9	372438191.3	215535236.8	219281076.1	227906282.5	394486598.3	391900888.9	404703214.3	372580643.7	341605639	335348213.6
phosphocholine	10145696160	9900376136	9107465337	6074999698	6159162457	6876132720	10567263278	9693888572	9497871592	8128064769	8107670531	7615077208
phosphocreatine	1169677.847	1156558.048	1052340.913	1094654.641	1137071.808	1431292.926	1087613.669	1155109.782	1278223.804	808580.7753	794677.8463	849644.1865
phosphoenolpyruvate	941697.5489	770326.5219	785555.9663	325343.7894	429444.24	491513.6924	1124266.642	1117074.491	1047125.16	1612440.433	1845287.531	1542804.391
proline	2614819121	2270001453	2639305176	2390016550	2448745565	2456431085	2391063555	2463529340	2644034512	2877937228	2795392017	2861360811
propionylcarnitine	4729920.269	4653310.851	4856899.331	1935487.889	2285826.948	2287048.596	4877416.857	5101931.242	5128780.354	4558269.449	4252571.096	4913290.407
Pyridoxine	52399487.28	58880243.14	56278532.22	44964143.12	44903141.67	45197240.22	55540858.16	53179632.1	53555030.82	52291197.15	49029240.37	56245669.29
S.Adenosyl.L.Homocysteine	8496573.054	8721381.437	7739062.54	5427614.477	5786733.726	5280485.582	8480744.855	7874356.251	7366975.552	8075474.394	7541759.505	6377171.356
S.Adenosyl.L.methionine	131233729.7	121848853.4	128141064.7	102318278.4	96848092.28	93191304.1	135845079.7	132110052.4	127047815.4	116993222.8	116280435.5	117499330.5
sarcosine	97195224.97	96228108.45	90729483.59	87612800.08	76047247.75	85710727.34	96651415.38	94892775.69	83368042.86	81937053.51	81434462.48	73804582.46
Sedoheptulose.7.phosphate	1782617.907	1574140.182	1724055.87	6233960.134	7082385.867	5821319.182	1381008.884	2083474.69	1872198.83	5336931.682	5073797.166	5006343.707
serine	45788480.42	48565828.63	38121434.12	45923648.3	41523643.73	35370086.98	48184426.82	34589454.3	30476327.12	34638440.28	38041635.62	28446461.53
succinate	18899261.81	18898609.79	16215808.01	43944424.54	50349156.9	48920271.29	16388218.91	17036645.89	16389986.51	18982336.14	19582035.31	18455774.03
taurine	146642093.9	150256866.1	145886265.1	149239539.3	149356543.2	147503159	162225654.6	174168618.4	166534045.6	165125633.9	159200756.5	164191791.8
threonine	109309917.9	104693969.3	105134049.7	92330807.45	82356069.05	106437937.5	122033504.8	109728442.9	109055934.9	93825708.46	96641888.33	98803936.7
tryptophan	109515637.7	113755924.4	107727236.5	91104374.14	92489661.92	115376264.9	111734780.3	107305364.6	113277013.2	105411065.2	95111601.4	110994280.2
tyrosine	210020739.9	219236758.5	210733655.6	164718750	167304555.6	182403049.8	225137121	207050026.5	199534997.1	188295385.3	183023573.4	172989997.1
UDP.GlcNAc	48763747.7	46721317.82	45752261.91	15570920.77	14819355.73	14865735.76	49028609.44	46046123.67	47265187.73	31659248.99	30475038.77	29193128.82
UDP	7541706.755	7534167.904	7269902.773	5796274.81	5080263.822	5286845.537	6066309.354	5842207.763	5826181.836	5246478.607	5027136.995	5336824.906
UMP	712560.6089	599223.9171	843451.8206	1587487.908	1432629.869	875003.8416	416148.5719	574155.5526	580718.8213	308712.5224	392704.9299	538396.6883
Ureidosuccinic_acid	16899278.9	16531477.62	12274708.08	147515224.7	162027867.7	154316742.9	671198.7009	380945.5898	1053810.188	3350857.111	1922488.814	2425518.249
uric.acid	3088345.739	3660905.289	3736914.011	1911167.148	1942601.072	1666816.436	3012118.591	3283536.026	4227413.643	2676287.946	2480489.438	3668260.005
UTP	66932403.85	63228591.81	72256042.41	21287043.47	21325643.98	13058369.75	53453533.09	74686811.3	59661895.28	56182290.84	53854603.55	50465935.36
dihydroorotate	2091722.171	2099457.51	2000611.358	18869886.07	20494873.62	19569214.92	56538.84294	141844.5073	157907.2571	322413.3334	228738.3109	384084.077
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
2-hydroxyglutarate
3-methyl.2.oxopentanoic_acid
3-phosphoglycerate
Acetyl.CoA
acetylcarnitine
ADP
alanine
Alpha.Ketoglutarate
AMP
arginine
argininosuccinate
ascorbate_unc
asparagine
aspartate
asymmetric.dimethylarginine
ATP
beta.alanine
betaine
Butyrylcarnitine
C5.Carnitines
carnitine
carnosine
CDP.choline
CDP
cis.aconitate
citrate
citrulline
CMP
creatine
creatinine
CTP
Cys_Ell
cystathionine
cytidine
dihydroxyacetone_phosphate
dTMP
F6P_or_G1P
FAD
Fructose.1.6.bisphosphate
fumarate
gamma.aminobutyric_acid
GDP
Glucose.6.phosphate
glutamate
glutamine
glyceraldehyde_3.phosphate
glycerate
Glycerol.3.Phosphate
Glycerophosphocholine
glycine
GMP
GSH_Ellmans
GSSG
GTP
histidine
hypotaurine
IMP
isoleucine
lactic_acid
lysine
mannose
Methionine.sulfoxide
methionine
methylthioadenosine
myo.inositol
N.acetylglutamate
NAD.
NADH
NADP
NADPH
niacinamide
O.phosphoethanolamine
ornithine
palmitate
Pantothenate
phosphocholine
phosphocreatine
phosphoenolpyruvate
proline
propionylcarnitine
Pyridoxine
S.Adenosyl.L.Homocysteine
S.Adenosyl.L.methionine
sarcosine
Sedoheptulose.7.phosphate
serine
succinate
taurine
threonine
tryptophan
tyrosine
UDP.GlcNAc
UDP
UMP
Ureidosuccinic_acid
uric.acid
UTP
dihydroorotate
METABOLITES_END
#END