#METABOLOMICS WORKBENCH alanyl_20250326_083011 DATATRACK_ID:5785 STUDY_ID:ST003825 ANALYSIS_ID:AN007477 PROJECT_ID:PR002390
VERSION             	1
CREATED_ON             	December 17, 2025, 1:38 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                   	pyruvate and uridine treatment
ST:STUDY_SUMMARY                 	To investigate whether the effects of SLC31A1 knockout can be rescued with
ST:STUDY_SUMMARY                 	pyruvate and uridine treatment, SEM cells containing a SLC31A1-targeting sgRNA
ST:STUDY_SUMMARY                 	(guide #2) or a intergenic -targetting sgRNA (i.e. a control sgRNA,
ST:STUDY_SUMMARY                 	"Intergenic1") and Cas9 were FACS-purified based on SLC31A1 expression, and then
ST:STUDY_SUMMARY                 	treated with pyruvate and uridine for 48 hours. Cells were then cultured in
ST:STUDY_SUMMARY                 	RPMI-1640 containing 10% FBS and penicillin and streptomycin in 6-well plates,
ST:STUDY_SUMMARY                 	and then collected for metabolomics. This demonstrated that the changes in
ST:STUDY_SUMMARY                 	aspartate, ureidosuccinic acid/carbamoyl aspartate and nucleotides were largely
ST:STUDY_SUMMARY                 	reverted by treatment with pyruvate and uridine.
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           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1330	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:Vehicle	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1330.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1331	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:Vehicle	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1331.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1332	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:Vehicle	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1332.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1333	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:PyruvateAndUridine	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1333.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1334	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:PyruvateAndUridine	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1334.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1335	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:PyruvateAndUridine	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1335.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1336	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:Vehicle	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1336.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1337	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:Vehicle	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1337.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1338	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:Vehicle	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1338.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1339	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:PyruvateAndUridine	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1339.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1340	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:PyruvateAndUridine	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1340.raw
SUBJECT_SAMPLE_FACTORS           	-	20241123_QE2_HILIC_KO_PU_repeat_AYW1341	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:PyruvateAndUridine	RAW_FILE_NAME(raw file name)=20241123_QE2_HILIC_KO_PU_repeat_AYW1341.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 × 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	20241123_QE2_HILIC_KO_PU_repeat_AYW1330	20241123_QE2_HILIC_KO_PU_repeat_AYW1331	20241123_QE2_HILIC_KO_PU_repeat_AYW1332	20241123_QE2_HILIC_KO_PU_repeat_AYW1333	20241123_QE2_HILIC_KO_PU_repeat_AYW1334	20241123_QE2_HILIC_KO_PU_repeat_AYW1335	20241123_QE2_HILIC_KO_PU_repeat_AYW1336	20241123_QE2_HILIC_KO_PU_repeat_AYW1337	20241123_QE2_HILIC_KO_PU_repeat_AYW1338	20241123_QE2_HILIC_KO_PU_repeat_AYW1339	20241123_QE2_HILIC_KO_PU_repeat_AYW1340	20241123_QE2_HILIC_KO_PU_repeat_AYW1341
Factors	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:Vehicle	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:Vehicle	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:Vehicle	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:PyruvateAndUridine	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:PyruvateAndUridine	Sample source:SEM leukemia cells | sgRNA:Intergenic1 | treatment2:PyruvateAndUridine	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:Vehicle	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:Vehicle	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:Vehicle	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:PyruvateAndUridine	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:PyruvateAndUridine	Sample source:SEM leukemia cells | sgRNA:SLC31A1 guide #2 | treatment2:PyruvateAndUridine
2-hydroxyglutarate	843875718.2	1133072907	1129186207	927507527.7	1074938697	1255430012	1076490369	937369775.5	1429007632	814628223.2	1142289701	1072046124
3-phosphoglycerate	4773860.478	5241658.52	4936052.263	4582906.638	5892092.743	5465364.786	4445021.094	3773783.046	3771876.399	4649855.082	4961402.648	4846102.141
Acetyl.CoA	5165333.239	5086051.428	4682141.677	4157283.883	4043626.711	3800653.215	3612337.611	3518822.707	4196622.631	3995504.673	4151531.692	4097168.12
acetylcarnitine	45453395.27	47115878.78	43069207.36	58692521.04	58003363	63069511.41	46755374.24	52198370.73	52473576.82	77078315.46	80075099.35	80104231.12
ADP	443567072.3	405514368.4	411675212.1	362556519.5	312956303.9	317895052.9	340543586.5	338033082.2	334979436.3	340189393	293714231.7	309163154.4
alanine	66347235.3	69314721.08	64330120.52	139205769.6	125771353.6	136540816.4	89069979.33	83953775.52	87294253.1	147879725.2	161458692.3	130098077.6
arginine	1166993477	1102086277	1191620740	1204581970	1125004598	1092204498	1013374982	938604486.5	928200019.6	1110707011	1016181678	1068422145
argininosuccinate	10474704.45	10124301.49	10225926.88	12146990.52	11873807.94	11467729	13293190.51	13235792.28	12142142.73	12566048.01	12273478.77	12506026.36
asparagine	492974063.8	511693482.5	545727497.7	489282743	483880519.7	508118471.5	423739118.1	430710786.3	478886371.6	405318425.1	376370491.5	397112456.7
aspartate	21595838.49	18617094.61	22574247.55	20786353.55	20523172.18	23101305.62	7673384.787	6914558.539	8156591.982	19024852.52	16511485.61	16242612.6
asymmetric.dimethylarginine	6735180.272	6491494.762	6053032.714	6623377.506	6543870.434	6154476.735	5734396.153	5550069.436	5409400.526	5829116.056	5731490.493	5744606.849
beta.alanine	17671524.8	20307006.09	18974370.25	28339205.83	23777378.79	27467695.25	29528266.35	28230524.7	30921330.96	32392629.06	27529915.9	34840104.17
Butyrylcarnitine	35646095.02	32892775.96	32730226.41	37895292.68	41468523.68	27232918.53	33831252.94	39242947.93	40469996.36	38553804.56	37328654.28	36989041.17
C5.Carnitines	4186441.451	3124844.536	4205137.286	3271196.38	2854532.277	2116063.726	12706405.81	15927995.81	15827033.89	8204292.075	6626278.121	7549869.597
carnitine	107653884.3	110261505.5	107533033.6	83559762.53	84368637.74	82573471.68	122101870	119689483	122844354.3	106732982.8	103378953.5	95531467.23
carnosine	921822.8994	790837.2657	955882.9116	618567.7848	611490.5805	692525.2922	966641.5024	746971.5435	828090.7498	1179906.023	1369414.154	1097627.712
CDP	30432764.71	28701327.11	26466632.74	41793554.92	42782558.83	41649777.35	18414825.35	19287755.28	21326379.4	32459149.21	33347582.76	34772609.07
cis.aconitate	22647557.47	24359010.54	27278814.97	28400222.68	27316814.73	28312340.27	19856895.01	21176147.35	21324895.38	20103620.76	20783625.46	21671167.83
citrate	291504290.1	278353442.5	263861826.7	371588882	380955667.2	367043842.2	200546160.3	223655481.2	178704436.9	273739703.4	308092762.6	287189859.4
citrulline	14936508.36	16705449.74	15764439.63	16502497.65	15753849.05	16525332.67	14177923.53	14677964.41	17155895.32	15017246.22	15379211.65	13454097.37
creatine	224434437.9	242834615.1	230965256.9	214556066.7	210584788.4	193560979.6	196668101.4	204070499.8	202177081.1	210467426	201161619.2	191408292.9
creatinine	16953840.65	16942014.34	16694413.38	16860290.61	17695497.6	15318522.31	16424860.25	16581958.91	17261639.41	19088838.86	17875690.52	19075033.51
Cysteine_Ellmans_conjugated	10262903.39	9498037.074	10452767.88	12208879.02	9854397.002	8911440.137	10708624.71	8359619.321	9539074.713	15365394.8	15982023.41	15033143.85
cystathionine	5806135.729	6410273.621	6358787.141	5480469.449	5895420.487	5910520.04	4748144.014	5558483.955	6002087.966	5560105.304	6092478.05	6161820.322
dihydroxyacetone_phosphate	7155676.878	7435691.039	6547141.467	3783330.208	4359830.706	3951723.703	14889125.99	14802339.22	13365697.08	8555300.788	9140781.829	8897737.164
FAD	1399002.444	1562931.663	911753.6369	1139325.933	1226532.121	1262461.081	1059438.463	1086609.626	1058801.755	1185553.786	1200180.087	979411.4769
Fructose.1.6.bisphosphate	1323782.959	1453469.509	1312159.077	1460999.054	1330304.885	1328260.484	959346.4409	886806.705	1007522.978	1153064.394	1061356.801	1252204.733
fructose	32115398.69	32099911.34	32015106.45	32692160.58	34798478	37178493.52	28832375.81	32386583.68	33053716.62	27944418.84	26119282.44	27035992.1
fumarate	23873722.91	22685838.06	21768586.48	26413393.64	23458530.37	25195622.51	28580591.64	30015696.83	24144025.84	26246979.42	30151699.86	30173443.88
GDP	40868325.06	37419254.36	37756758.1	36323354.3	33301562.71	33464362.55	41993386.67	41115753.93	40213435.74	34102052.9	31278733.58	30953114.75
glutamate	971307314.6	904269150.6	945880560.3	818073685.8	770934888.8	782806846.7	654417620.6	617762028.8	675398343.9	730217902.4	634510109.4	657076848.5
glutamine	2017542389	1962638390	2097385916	2133165961	1966610034	2122171488	1105167171	1123627977	1280358778	1874709895	1565670649	1632587349
glycerate	659176468.8	703458804	693264530.8	745086986.1	867829248.8	844816899.6	678364108.3	648119926.3	719960103.2	622061879.9	614745348.9	599602533.7
Glycerol.3.Phosphate	15029345.27	17101129.46	14123252.66	2838920.955	2593879.51	2212208.233	31772447.92	33616702.16	36479222.05	6972594.937	6757898.016	6758023.139
Glycerophosphocholine	2179435590	2310857772	2359080850	1979090931	1868827208	2042761979	946381882.2	992523051	1142927275	1263979184	1142947103	1184966446
glycine	10869273.76	10035809.85	9820181.977	10697808.19	9497153.111	11127019.65	8434363.153	8735172.436	9293003.686	10459636.53	9610298.131	10494598.9
GSH_Ellmans	299932571.8	296340206.7	296555105.5	277050792.5	261560670.5	274825446.2	240218120.7	256455925.9	260778181.9	268686410.2	249163924.3	253545373.8
GSSG	5803256.024	5094950.765	4853967.069	4555031.287	3979021.56	3934546.414	4257629.547	3875766.16	3799768.219	4347666.886	3712306.487	3972397.614
histidine	238849424.2	237954090.4	239414877.6	227068081.4	205351895.3	220270790.1	200729188.9	200320866.6	209134321.8	214302501.5	201545500.7	201727258.7
homocitrulline	1211947.951	1186550.704	1221500.961	1044112.596	1197758.867	913170.2941	1285241.435	1270466.665	930894.7786	1283049.789	1192286.148	1012867.699
hypotaurine	15432546.74	18615900.06	20526806.2	18023291.71	20296829.8	25729071.58	20224095.06	23557423.56	23733122.48	15821373.65	19081380.35	20946072.21
IMP	1305833.296	1459027.379	1136691.789	693662.8301	824586.9044	690748.9114	6285674.061	5995135.351	4955553.93	947786.1751	1630050.183	1481302.849
isoleucine	347945847.7	366048605.7	367017056.2	229016186.6	253131365.6	284243863.9	299786718.3	264824838.8	306070604.8	258912851.2	219483195	277290542.6
kynurenine	2418490.611	1883855.516	3190721.315	2271814.765	2007910.743	2009202.043	2817690.436	3116100.263	2830328.402	3543520.065	3182078.836	3197951.466
leucine	277078102.3	205612432.7	367017056.2	164054441.8	165123165.7	284243863.9	196844261.5	223075927.1	215242125.9	169853973	180876440.7	149490867
lysine	10695701.66	10466049.73	11238545.3	10246858.08	10106142.76	10343025.17	9392372.38	8434453.549	8716527.531	11853224.96	12126224.26	11075617.61
malate	322335903.4	298772121	294938452.4	367472954.4	362789103.7	361785798.3	349644041.8	376710768.1	357725307.7	392074079.8	376963906.9	382418943.5
mannose	32115994.53	32034675.08	32110979.59	32217305.86	34733350.11	37108398.59	28834556.81	32385093.36	32758678.28	27797937.05	26061834.4	27035303.74
Methionine.sulfoxide	80043084.2	77971010.01	74976866.18	76837711.42	65121772.62	69123597.39	64290286.22	59300370.04	64465358.69	68339410.95	64122524.95	61727623.64
methionine	108211185.3	111869092.3	118931206.4	99900739.7	96459870.8	103262192.6	90003418.85	79416381	80347694.67	104223108.7	76908467.42	95728818.67
methylthioadenosine	4659223.353	4398395.994	3620702.272	4397068.768	5616491.322	3644260.795	3074721.664	4480408.442	4241789.297	4244427.838	5048088.74	4600439.706
NAD.	74562654.54	76851046.46	74142428.38	77165348.89	75720435.13	75088163.63	61241332.98	63088861.12	63507938.05	73815901.03	71589979.42	75438291.5
NADH	12443718.37	14057043.16	13848009.75	8849434.847	8028894.908	6669617.546	18577343.61	17966745.28	17915510.11	11796829.97	10224832.84	10037701.64
NADP	927549.4837	807124.6162	793586.6522	658201.6773	701423.9394	690639.5402	641451.8017	658473.944	635445.2995	659827.8451	659040.5817	654251.3147
NADPH	4070652.444	4118966.616	3984679.177	4095023.224	3804010.934	3716278.733	3379749.517	3197928.191	3163439.552	3317601.963	3128158.446	3223894.255
niacinamide	15631449.3	15351500.76	14723224.24	18460906.84	25695650	20840898.34	16031402.01	19512250.81	18890547.35	25755169.57	27389656.42	28696192.57
ornithine	16113980.42	15797517.06	17431748.48	15578600.8	14273717.11	14474307.56	11792217.4	12335999.91	11797645.15	15470299.17	14441455.95	14991366.16
palmitate	151706663.7	113674117.8	95449522.57	88450482.56	108687641.6	97601942.73	140073743.7	67854498.45	96062039.73	59719407.1	88595635.25	126538155.3
phosphocholine	11479850580	12204482764	11801797629	9821369043	9830083186	9995602439	7689508468	8226464168	9438699397	7838696322	8159761144	7879159599
phosphocreatine	941693.3609	686541.6116	982781.2515	984181.4691	421054.1194	592701.8267	729591.4424	640269.0623	693171.7241	619028.7032	503075.0144	715916.3229
phosphoenolpyruvate	949449.1673	1049699.764	932938.1138	1146514.331	1580884.451	1623245.459	538159.8212	378499.0139	405013.9058	1110174.956	1121842.559	1165389.249
proline	2374987584	2010299159	2145004215	1826080168	1730267900	1742941142	2503037016	2147445348	2374646007	2056281886	2050388426	1958863259
propionylcarnitine	1916234.258	1894289.68	1578997.119	2446104.871	2343238.446	2100435.662	1509808.08	1430607.271	1706119.175	2254846.123	2393405.621	2234104.09
Pyridoxine	32762270.95	35809094.59	31730676.9	35261775.7	40008232.93	29651507.83	32031576.69	34440286.51	35673412.24	37488622.83	38273805.37	38381121.02
S.Adenosyl.L.Homocysteine	5144810.243	5315829.882	5212553.456	5649394.604	5861976.775	5274098.574	4078643.737	3151413.236	4115667.694	4964939.2	5255094.547	5216935.169
S.Adenosyl.L.methionine	59045373.86	57717600.23	59026889.8	59072055.93	55396802.3	56020434.65	49793679.3	50594372.43	51561627.31	51676215.39	51936712.12	50068026.85
sarcosine	64774804.99	69314721.08	64330120.52	139205769.6	126238158	136540816.4	89069979.33	84141172.34	88151775.6	147464164.3	161458692.3	130209213.6
serine	29034461.55	31141858.7	33753013.21	35317194.86	34370571.37	41261472.29	22983434.17	23352958.52	25880924.09	38306214.5	36604976.04	38862779.15
succinate	32582774.31	46288888.27	42713141.8	30385920.99	37710247.01	43023982.16	63483125.37	59635267.29	60270940.85	51595253.73	56777817.44	56544446.07
threonine	77476175.88	84608182.5	88849600.2	85689745.86	89899537.03	83735057.65	67612527.29	54978567.92	70345544.09	82527263.21	76250997.37	83673927.6
trans.urocanate	8900187.217	16033002.58	14313333.27	13369739.41	10748102.35	12909193.5	7911053.01	10888229.35	13088203.87	8102786.65	14311433.27	9092766.696
tryptophan	38430640.4	42412254.63	41534806.46	36398038.77	36711599.29	39966773.14	33397535.22	30807830.99	29899680.32	40053417.98	30418421.66	38428363.67
tyrosine	154948882.2	164265685.9	163952390.5	145757345.5	146978372.6	145260866.5	132640244.8	98325494.16	130903136	129167985.4	131945958.9	129503693.5
UDP.GlcNAc	47618103.8	46040738.91	45633785.23	95054163.33	92327934.74	93683710.66	25227505.69	26356081.18	28865164.42	69261010.66	69294747.77	69042483.79
UDP	138962732.8	131079479.1	128177081.4	238237166.6	225881047.6	228356975.1	90343745.3	90483651.93	94792669.39	177311111.9	178329801.6	182155562.9
Carbamoyl Aspartic Acid	7859815.966	7937800.044	7534437.861	NA	NA	NA	18014436.16	17939344.94	20463767.95	1788968.086	1249602.448	1214449.755
xanthosine	3600776.109	3637358.07	4063537.294	3284547.83	3777739.751	3997540.626	3110359.608	3829469.95	3413491.427	3107084.091	2785528.947	3002764.694
dihydroorotate	1056783.736	980695.1	613395.9468	107213.7734	41127.85367	115141.5891	2040926.129	1687718.241	2139903.772	37520.23264	100886.5289	0
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
2-hydroxyglutarate
3-phosphoglycerate
Acetyl.CoA
acetylcarnitine
ADP
alanine
arginine
argininosuccinate
asparagine
aspartate
asymmetric.dimethylarginine
beta.alanine
Butyrylcarnitine
C5.Carnitines
carnitine
carnosine
CDP
cis.aconitate
citrate
citrulline
creatine
creatinine
Cysteine_Ellmans_conjugated
cystathionine
dihydroxyacetone_phosphate
FAD
Fructose.1.6.bisphosphate
fructose
fumarate
GDP
glutamate
glutamine
glycerate
Glycerol.3.Phosphate
Glycerophosphocholine
glycine
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
serine
succinate
threonine
trans.urocanate
tryptophan
tyrosine
UDP.GlcNAc
UDP
Carbamoyl Aspartic Acid
xanthosine
dihydroorotate
METABOLITES_END
#END