#METABOLOMICS WORKBENCH cendalfr_20240411_071306 DATATRACK_ID:4769 STUDY_ID:ST003166 ANALYSIS_ID:AN005195 PROJECT_ID:PR001970
VERSION             	1
CREATED_ON             	April 11, 2024, 1:33 pm
#PROJECT
PR:PROJECT_TITLE                 	Releasing the mitochondrial respiration brake MCJ/DnaJC15 enhances CD8 CAR-T
PR:PROJECT_TITLE                 	cell therapy efficacy
PR:PROJECT_SUMMARY               	Metabolism of chimeric antigen receptor (CAR) T cells is emerging as an
PR:PROJECT_SUMMARY               	important area to improve CAR-T cell therapy in cancer treatment. Mitochondrial
PR:PROJECT_SUMMARY               	respiration is essential for survival and function of CAR-T cells, but
PR:PROJECT_SUMMARY               	developing strategies to specifically enhance mitochondrial respiration has been
PR:PROJECT_SUMMARY               	challenging. Here we identify MCJ/DnaJC15, an endogenous negative regulator of
PR:PROJECT_SUMMARY               	mitochondrial Complex I, as a metabolic target to enhance mitochondrial
PR:PROJECT_SUMMARY               	respiration in CD8 CAR-T cells. Loss of MCJ in CD8 CAR-T cells increases their
PR:PROJECT_SUMMARY               	in vitro and in vivo efficacy against mouse B cell leukemias. MCJ deficiency in
PR:PROJECT_SUMMARY               	TCR- specific CD8 cells also increases their efficacy against solid tumors in
PR:PROJECT_SUMMARY               	vivo. Furthermore, we reveal that human CD8 cells express MCJ and that silencing
PR:PROJECT_SUMMARY               	MCJ expression increases mitochondrial metabolism and anti-tumor activity of
PR:PROJECT_SUMMARY               	human CAR-T cells. Thus, targeting MCJ to enhance mitochondrial metabolism is a
PR:PROJECT_SUMMARY               	promising therapeutic strategy to improve the efficacy of adoptive T cell
PR:PROJECT_SUMMARY               	therapies.
PR:INSTITUTE                     	University of Colorado School of Medicine
PR:LABORATORY                    	Laboratory of Angelo D'Alessandro in collaboration with Mercedes Rincon
PR:LAST_NAME                     	Cendali
PR:FIRST_NAME                    	Francesca
PR:ADDRESS                       	13199 East Montview Boulevard, Aurora, CO, 80045, USA
PR:EMAIL                         	francesca.cendali@cuanschutz.edu
PR:PHONE                         	3037246131
#STUDY
ST:STUDY_TITLE                   	Metabolomics of Murine WT or MCJ KO CD19-BBz CD8 CAR-T cells
ST:STUDY_SUMMARY                 	MCJ/DnaJC15 is an endogenous negative regulator of Complex I and mitochondrial
ST:STUDY_SUMMARY                 	respiration. The goal of these experiments are to characterize the metabolic
ST:STUDY_SUMMARY                 	profile of murine WT or MCJ KO CD8 CD19-BBz CAR-T cells after 3 expansions (6
ST:STUDY_SUMMARY                 	days) with IL-2 (60IU/ml).
ST:INSTITUTE                     	University of Colorado School of Medicine
ST:LABORATORY                    	Laboratory of Angelo D'Alessandro in collaboration with Mercedes Rincon
ST:LAST_NAME                     	Cendali
ST:FIRST_NAME                    	Francesca
ST:ADDRESS                       	13199 East Montview Boulevard, Aurora, CO, 80045, USA
ST:EMAIL                         	francesca.cendali@cuanschutz.edu
ST:PHONE                         	3037246131
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Mus musculus
#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           	-	WT CD19-BBz CART_1	factor:WT | Sample source:T-Cells	RAW_FILE_NAME(Raw File)=1
SUBJECT_SAMPLE_FACTORS           	-	WT CD19-BBz CART_2	factor:WT | Sample source:T-Cells	RAW_FILE_NAME(Raw File)=2
SUBJECT_SAMPLE_FACTORS           	-	WT CD19-BBz CART_3	factor:WT | Sample source:T-Cells	RAW_FILE_NAME(Raw File)=3
SUBJECT_SAMPLE_FACTORS           	-	WT CD19-BBz CART_4	factor:WT | Sample source:T-Cells	RAW_FILE_NAME(Raw File)=4
SUBJECT_SAMPLE_FACTORS           	-	MCJ KO CD19-BBz CART_1	factor:MCJ KO | Sample source:T-Cells	RAW_FILE_NAME(Raw File)=5
SUBJECT_SAMPLE_FACTORS           	-	MCJ KO CD19-BBz CART_2	factor:MCJ KO | Sample source:T-Cells	RAW_FILE_NAME(Raw File)=6
SUBJECT_SAMPLE_FACTORS           	-	MCJ KO CD19-BBz CART_3	factor:MCJ KO | Sample source:T-Cells	RAW_FILE_NAME(Raw File)=7
SUBJECT_SAMPLE_FACTORS           	-	MCJ KO CD19-BBz CART_4	factor:MCJ KO | Sample source:T-Cells	RAW_FILE_NAME(Raw File)=8
#COLLECTION
CO:COLLECTION_SUMMARY            	Mouse CD8 cells were isolated with negative selection as described above,
CO:COLLECTION_SUMMARY            	activated with anti-CD3/anti-CD28 beads (Dynabeads™ Mouse T-Activator CD3/CD28
CO:COLLECTION_SUMMARY            	for T-Cell Expansion and Activation, Gibco, Cat#11453D) with addition of
CO:COLLECTION_SUMMARY            	recombinant human IL-2 (rhIL-2) (60 IU/ml) and human recombinant IL-7 (10
CO:COLLECTION_SUMMARY            	ng/ml). On the second and third day of activation, the supernatant containing
CO:COLLECTION_SUMMARY            	the CD19-BBz CAR/hEGFR retrovirus was spun on a plate coated with retronectin
CO:COLLECTION_SUMMARY            	(Takara Bio Inc.) (2000xg for 2.5h at 32C). The activated cells were added to
CO:COLLECTION_SUMMARY            	the viral-coated plate for transduction. The transduced CD8 CAR-T cells were
CO:COLLECTION_SUMMARY            	removed from the beads on the fourth day and expanded with rhIL-2 (60 IU/ml) or
CO:COLLECTION_SUMMARY            	rhIL-7 (10 ng/ml) and rhL-15 (100 ng/ml). After 2 days (1st expansion) cells
CO:COLLECTION_SUMMARY            	were split 1/3 with fresh cytokine-containing medium. After 2 more days (2nd
CO:COLLECTION_SUMMARY            	expansion), cells were split again with fresh cytokine-containing medium. Two
CO:COLLECTION_SUMMARY            	days later (3rd expansion) cells were used for experiment or incubated in
CO:COLLECTION_SUMMARY            	cytokine free-medium. For CAR+ cells enrichment, the CAR-T cells expanded with
CO:COLLECTION_SUMMARY            	IL-2 for 3 expansions were incubated with an anti-hEGFR-PE (BioLegend,
CO:COLLECTION_SUMMARY            	Cat#352904, RRID: AB_10896794) antibody, followed by incubation with anti-PE
CO:COLLECTION_SUMMARY            	microbeads (Miltenyi, order#130-048-801), and purified using the Miltenyi LS
CO:COLLECTION_SUMMARY            	columns as recommended by the manufacturer (Miltenyi). The enrichment resulted
CO:COLLECTION_SUMMARY            	in a 98-100% CAR+ population, as verified by flow cytometry.
CO:SAMPLE_TYPE                   	T-cells
#TREATMENT
TR:TREATMENT_SUMMARY             	Metabolomics differences in this experiment are observed at baseline
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	CAR-T cells were isolated as described above either from in vitro culture or
SP:SAMPLEPREP_SUMMARY            	from bone marrow harvested in an in vivo study. The cells were washed in PBS and
SP:SAMPLEPREP_SUMMARY            	frozen at -80C until the assay is ready to run. Metabolites from cells were
SP:SAMPLEPREP_SUMMARY            	extracted at 2x106 cells/ml at 4°C (30 min) in the presence of 5:3:2
SP:SAMPLEPREP_SUMMARY            	MeOH:MeCN:water (v/v/v). The samples were spun down and the resulting
SP:SAMPLEPREP_SUMMARY            	supernatant was transferred to new tubes and dried under a vacuum. The resulting
SP:SAMPLEPREP_SUMMARY            	residue was reconstituted in 0.1% formic acid at a 3x concentration, then
SP:SAMPLEPREP_SUMMARY            	analyzed on a Thermo Vanquish UHPLC coupled to a Thermo Q Exactive MS as
SP:SAMPLEPREP_SUMMARY            	previously described in detail.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Positive Ion Mode
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
CH:SOLVENT_A                     	100% water; 0.1% formic acid
CH:SOLVENT_B                     	100% acetonitrile; 0.1% formic acid
CH:FLOW_GRADIENT                 	0-0.5 min 5% B, 0.5-1.1 min 5-95% B, 1.1-2.75 min hold at 95% B, 2.75-3 min
CH:FLOW_GRADIENT                 	95-5% B, 3-5 min hold at 5% B.
CH:FLOW_RATE                     	0.450ml/min
CH:COLUMN_TEMPERATURE            	45
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms,
MS:MS_COMMENTS                   	microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV,
MS:MS_COMMENTS                   	capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas
MS:MS_COMMENTS                   	0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were
MS:MS_COMMENTS                   	annotated and integrated using Maven in conjunction with the KEGG database.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Peak Area
MS_METABOLITE_DATA_START
Samples	WT CD19-BBz CART_1	WT CD19-BBz CART_2	WT CD19-BBz CART_3	WT CD19-BBz CART_4	MCJ KO CD19-BBz CART_1	MCJ KO CD19-BBz CART_2	MCJ KO CD19-BBz CART_3	MCJ KO CD19-BBz CART_4
Factors	factor:WT | Sample source:T-Cells	factor:WT | Sample source:T-Cells	factor:WT | Sample source:T-Cells	factor:WT | Sample source:T-Cells	factor:MCJ KO | Sample source:T-Cells	factor:MCJ KO | Sample source:T-Cells	factor:MCJ KO | Sample source:T-Cells	factor:MCJ KO | Sample source:T-Cells
L-alanine	1560974	1860581	1569585	1809974	1637607	2132121	2116687	2287936
L-arginine	9474497	9690346	8696651	8866196	9816144	11296820	8652307	10939020
L-asparagine	366180.8	448378.4	323211.7	369701.5	278499.3	449202.2	470385.2	499535.5
L-aspartate	2126716	2682148	1893171	2323962	1772503	2600101	2738762	3014410
L-cysteine	36903.59	39842.39	33297.95	33459.18	29179.15	42591.98	35181.66	42745.12
L-glutamate	5148734	6169774	4316300	4601535	4209810	5663506	6202686	6222770
L-glutamine	1131900	1385852	943984.8	1057698	1101014	1477172	1297373	1405208
L-histidine	661184.2	501516.5	588026.3	698885.8	432550.2	546134.3	508735.3	559171.1
L-leucine/isoleucine	5859734	6118064	6318552	6438852	7540604	7634414	6560772	6775482
L-lysine	339298.1	343652.8	355650.7	408541.8	408594.7	331521.5	384127	390379.9
L-methionine	1628233	1278501	1627498	1538566	1840196	2034489	1925362	1788372
L-phenylalanine	2344554	2354135	2706233	3192963	2943106	2643999	2360224	2219698
L-proline	9453505	10765260	9427457	10355620	12292660	14385920	13907360	13842290
L-serine	750107.7	699896.8	603004.6	778602.2	386744.7	566277.7	637541.5	787477.4
L-threonine	410822.1	457620.2	366732.2	429815.2	271374.6	389165.1	429592.2	473229.5
L-tryptophan	1169436	946013.8	1171434	1162017	1207147	1209066	1166954	1255822
L-tyrosine	3590117	2120591	3372709	2435818	3792917	3816349	3004382	2874980
L-valine	5458792	6064960	6007768	5817142	6208846	6386062	5819806	6601522
L-cystine	19560.68	18091.37	18358.91	20575.74	23995.64	28793.09	23110.23	24356.81
ADP	192898.8	164063.2	151915	186493.9	377326.6	471596.1	613402.7	381522.7
AMP	35515510	37689240	40821240	35090290	36996660	47375260	45433750	55747920
dAMP	37690.01	43403.94	33439.31	45304.27	28630.04	23555.89	21198.91	31695.05
Adenosine	1914259	1080486	2395523	1323620	2353503	2402693	1510885	1581484
Adenine	2045558	1979711	2059359	1872918	2181581	2459223	2231389	2729003
GMP	5196508	4458117	5188418	4493158	3903462	5009770	3893602	5243548
Guanosine	99374.78	131166.4	100462.3	137092.4	127640.8	133580.2	130700.6	176768.9
Guanine	1397137	1260923	1377803	1264460	1182391	1424175	1105561	1550463
CMP	378148.1	346206.5	382826.3	366437.1	286132.8	403849.5	349496.7	487927.7
Cytidine	70830.42	64305.09	64315.22	59809.16	51369.25	82231.55	69420.44	124965.9
Cytosine	330017.3	279875.9	286526.6	260101	349136.8	358298.5	295816.5	476999.9
UDP	88846.99	80037.07	76711.02	78554.73	92282.38	103437.4	114533.4	121617
UMP	893351.7	801612.2	799320.7	664863.2	656865.8	927946.5	770534.5	1087464
Uracil	170400	113466.4	145534.8	127921.6	132097.4	130337.7	143446.6	175422.1
IMP	112426.1	63151.55	43833.18	66928.02	93931.57	55801.08	95920.73	122842.4
Inosine	260858.9	270459.9	206688.2	231771.1	234127.7	209468.8	214015.7	358613
Hypoxanthine	2586442	2251369	2179575	2058654	2064268	1661591	1906416	2635113
Xanthine	270286.6	211436.8	268019.8	230413	318553.8	296458.7	282762.5	350826.2
Allantoate	401791.5	266614.3	391021.5	414176.3	246724.8	275889.5	265316.6	292002.8
5-Hydroxyisourate	61375.47	75428.15	43838.48	76877.54	56298.69	86484.12	95102.95	90302.27
Urate	761466.3	754633.8	702935.4	767341.8	786828.8	857789.4	769502.1	712302.5
Nicotinamide	21011410	17360390	20466720	20197780	16338690	20371110	15552190	20396270
NADP+	5944.648	4702.077	3955.111	6155.706	13050.12	14033.09	18497.15	17892.24
NAD+	427988.1	611790.5	470638	590737.4	882142.1	910126.3	1119133	891170.4
Glutathione	67228420	61479730	64665080	61079530	82059820	69900010	88248770	63110770
Glutathione disulfide	333620.8	350272.5	439908.2	345591.3	504896.9	424291.7	487633.1	450908
5-Oxoproline	9190771	5751874	8419663	8360978	6828328	6147630	6911890	7221250
Cys-Gly	2045320	1810221	1934998	1624655	2580867	2235762	2676760	2037312
Ascorbate	306599.2	292930.4	252818.2	281781	253225.1	393053.6	283050.3	264809.5
Dehydroascorbate	309890	232302.8	285360.2	249096.5	380837.4	381146.1	340566.7	340860.3
gamma-L-Glutamyl-L-cysteine	183657.3	171131.7	206872.2	179775.6	252164	212365.3	278628.2	258832.5
gamma-Glutamyl-Se-methylselenocysteine	146896.1	121124.3	169048.4	160805.3	105142.3	115559.9	113414.1	115571
Phosphoserine	58135.73	57209.11	48172.98	64588.56	64139.77	68949.14	83072.52	64131.7
S-Adenosyl-L-homocysteine	80175.12	81068.87	112079	79766.98	104394.3	78656.77	98485.43	100371
S-Adenosyl-L-methionine	151653.6	151432.1	161634.1	175776.7	188482.3	156553.1	137030.6	177016.3
Ornithine	347157	219038	290127.9	367067.1	157608.4	206632.3	217007.8	232587.5
L-Citrulline	71532.2	70962.69	75121.22	78022.5	33514.62	57404.38	48709.61	43118.41
Argininosuccinate	83243.6	108403.5	62496.84	74438.84	53439	109909.8	117585.5	119640.6
Putrescine	60132	56292.78	131177.8	67566.41	132702.4	83762.62	64386.08	71458.27
Spermidine	200398	152133.4	333433.6	288685.4	284191.7	270282.8	325427.5	216473.2
Spermine	40694.37	24330.67	54350.11	64880.25	55700.58	49597.92	96055	59683.52
Homocarnosine	642766.9	679692.1	777400.5	1004225	578504.8	747302.4	1007291	966757.2
Creatine	5177788	6588714	4733909	4985474	5761802	6652140	6652768	7176420
Creatinine	877739.8	1099076	1062474	844304.8	1070186	1191026	572555.8	976383.8
N-Acetyl-L-citrulline	101746.6	148805.8	218326.1	153648.3	94885.44	248451.2	96684.23	84767.32
Guanidinoacetate	113637	128295.4	102793.8	127159.5	145252.1	163708.1	170739.6	182407.4
trans-4-Hydroxy-L-proline	688776.5	813576	604331	700445.5	651177.5	827619.2	812960.8	951099.3
N-Succinyl-L-glutamate 5-semialdehyde	25476.84	32440.41	26563.51	28664.74	48660.39	53142.71	54627.5	53622.96
Taurine	3990009	4621702	3696854	4335012	3985802	4890046	4682844	5102597
Hypotaurine	272966.5	321640.2	229491.9	253244.8	249120.2	314781	360660.5	340369.4
L-Methionine S-oxide	99382.91	66533.93	99130.65	58440.11	147734.5	88623.38	101842.4	100555.5
3-Methyleneoxindole	107182.1	81971.71	111309.6	121574.8	109759.1	116509.4	118826.9	122928.6
Indole-3-acetaldehyde	50228.39	53816.46	63512.91	61611.59	86200.88	83387.26	71885.32	62432.14
Indole-3-acetate	22670.97	25459.15	25484.29	24670.5	33807.91	26837.95	23201.48	28196.42
Indolepyruvate	68129.73	61251.06	76607.94	81111.93	84724.55	49792.39	82357.98	120409.6
Anthranilate	78141.65	76254.41	90995.01	251173.4	141108.7	130119.5	120866.8	121913.2
g-Oxalo-crotonate	54072.84	43222.35	47714.7	50868.53	38439.63	29265.34	23571.96	62519.31
Ectoine	176738	120542.9	162277.4	145142.7	396198.6	344297.6	302579	231293.7
Sphingosine	66541.25	100253.3	231344.9	193211.5	198160.3	174840.7	170430.5	206426.8
Acetylcholine	353667	368839.1	434279.3	387372.8	517535.2	522618.2	540154.7	562411.2
Choline	3224907	3885290	3862851	3901493	5203778	4727862	4532216	4229265
L-Carnitine	1346911	1248948	1488786	1356636	1796599	1848186	1387678	1430163
acetyl-carnitine (acyl-C2)	618760.9	609696.9	595332	679534.3	785650.8	770561.3	814609.2	706110.2
propionyl-carnitine (acyl-C3)	104950.9	144916.5	136092.4	157642.3	187646.9	225789.4	163514.9	159785
butanoyl-l-carnitine (acyl-C4)	50854.95	46403.22	50090.36	49512.27	74722.03	56788.99	57362.75	45174.44
hexanoyl-L-carnitine (acyl-C6)	25401.28	30309.29	38629.7	29021.29	40433.96	39697.02	59966	62707.16
L-octanoylcarnitine (acyl-C8)	11968.05	11860.19	12134.86	11483.64	16225.39	16382.98	22199.16	21193.7
O-Decenoyl-L-carnitine (acyl-C10:1)	23410.67	24079.27	29065.29	22997.48	22603.32	32466.45	29756.58	33181.12
O-dodecenoyl-carnitine (acyl-C12:1)	1421730	1980051	1862438	1734403	1486989	1249311	1613914	1657771
O-tetradecanoyl-L-carnitine (acyl-C14)	5699.241	2329.718	25782.49	13839.59	27347.76	25429.02	39969.91	38373.44
L-Palmitoylcarnitine (acyl-C16)	25785.27	25822.05	92625.81	75484.43	68576.52	59447.41	101208.5	103265.4
Hexadecenoyl-carnitine (acyl-C16:1)	11781.93	11879.79	31447.86	26438.68	29753.65	27310.72	24645.47	28878.34
Octadecanoyl-L-carnitine (acyl-C18)	24538.23	10639.32	80052.68	32577.5	22221.78	254316.7	25376.87	92576.46
O-octadecenoyl-L-carnitine (acyl-C18:1)	11460.83	11977.58	47281.33	28718.54	24955.74	25115.77	21565.84	29061.51
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	CmpdID	parent	medRt
L-alanine	C00041	90.06	0.684
L-arginine	C00062	175.12	0.650
L-asparagine	C00152	133.06	0.677
L-aspartate	C00049	134.04	0.675
L-cysteine	C00097	122.03	0.667
L-glutamate	C00025	148.06	0.680
L-glutamine	C00064	147.08	0.692
L-histidine	C00135	156.08	0.624
L-leucine/isoleucine	C00123	132.10	1.086
L-lysine	C00047	147.11	0.623
L-methionine	C00073	150.06	0.944
L-phenylalanine	C00079	166.09	1.634
L-proline	C00148	116.07	0.735
L-serine	C00065	106.05	0.674
L-threonine	C00188	120.07	0.679
L-tryptophan	C00078	205.10	1.776
L-tyrosine	C00082	182.08	0.971
L-valine	C00183	118.09	0.699
L-cystine	C00491	241.03	0.694
ADP	C00008	428.04	0.897
AMP	C00020	348.07	0.832
dAMP	C00360	332.08	0.939
Adenosine	C00212	268.10	0.978
Adenine	C00147	136.06	0.811
GMP	C00144	364.06	0.886
Guanosine	C00387	284.10	1.022
Guanine	C00242	152.06	0.901
CMP	C00055	324.06	0.713
Cytidine	C00475	244.09	0.790
Cytosine	C00380	112.05	0.798
UDP	C00015	405.01	0.819
UMP	C00105	325.04	0.829
Uracil	C00106	113.04	0.932
IMP	C00130	349.05	0.912
Inosine	C00294	269.09	1.066
Hypoxanthine	C00262	137.05	0.958
Xanthine	C00385	153.04	0.954
Allantoate	C00499	177.06	0.636
5-Hydroxyisourate	C11821	185.03	0.679
Urate	C00366	169.04	0.938
Nicotinamide	C00153	123.06	0.947
NADP+	C00006	744.08	0.855
NAD+	C00003	664.12	0.881
Glutathione	C00051	308.09	0.925
Glutathione disulfide	C00127	613.16	0.935
5-Oxoproline	C01879	130.05	0.963
Cys-Gly	C01419	179.05	0.940
Ascorbate	C00072	177.04	1.915
Dehydroascorbate	C05422	175.02	0.980
gamma-L-Glutamyl-L-cysteine	C00669	251.07	0.941
gamma-Glutamyl-Se-methylselenocysteine	C05695	313.03	0.637
Phosphoserine	C02532	186.02	0.683
S-Adenosyl-L-homocysteine	C00021	385.13	0.941
S-Adenosyl-L-methionine	C00019	399.14	0.657
Ornithine	C01602	133.10	0.625
L-Citrulline	C00327	176.10	0.684
Argininosuccinate	C03406	291.13	0.674
Putrescine	C00134	89.11	0.617
Spermidine	C00315	146.17	0.585
Spermine	C00750	203.22	0.580
Homocarnosine	C00884	241.13	0.933
Creatine	C00300	132.08	0.701
Creatinine	C00791	114.07	0.683
N-Acetyl-L-citrulline	C15532	218.11	0.678
Guanidinoacetate	C00581	118.06	0.682
trans-4-Hydroxy-L-proline	C01157	132.07	0.681
N-Succinyl-L-glutamate 5-semialdehyde	C05932	232.08	0.876
Taurine	C00245	126.02	0.703
Hypotaurine	C00519	110.03	0.681
L-Methionine S-oxide	C02989	166.05	0.944
3-Methyleneoxindole	C02796	146.06	1.802
Indole-3-acetaldehyde	C00637	160.08	1.222
Indole-3-acetate	C00954	176.07	1.851
Indolepyruvate	C00331	204.06	0.981
Anthranilate	C00108	138.05	0.742
g-Oxalo-crotonate	C03453	159.03	0.941
Ectoine	C06231	143.08	1.038
Sphingosine	C00319	300.29	1.986
Acetylcholine	C01996	146.12	0.714
Choline	C00114	104.11	0.669
L-Carnitine	C00318	162.11	0.683
acetyl-carnitine (acyl-C2)	C02571	204.12	0.892
propionyl-carnitine (acyl-C3)	C03017	218.14	1.137
butanoyl-l-carnitine (acyl-C4)	C02862	232.15	1.793
hexanoyl-L-carnitine (acyl-C6)	HMDB00756	260.19	1.832
L-octanoylcarnitine (acyl-C8)	HMDB00791	288.22	1.867
O-Decenoyl-L-carnitine (acyl-C10:1)	CID57357170	314.24	2.427
O-dodecenoyl-carnitine (acyl-C12:1)	HMDB13326	342.26	1.830
O-tetradecanoyl-L-carnitine (acyl-C14)	HMDB05066	372.31	1.977
L-Palmitoylcarnitine (acyl-C16)	C02990	400.34	2.030
Hexadecenoyl-carnitine (acyl-C16:1)	HMDB06317	398.33	2.008
Octadecanoyl-L-carnitine (acyl-C18)	HMDB00848	428.37	2.045
O-octadecenoyl-L-carnitine (acyl-C18:1)	HMDB06351	426.36	2.051
METABOLITES_END
#END