#METABOLOMICS WORKBENCH jravilap_20250728_115741 DATATRACK_ID:6222 STUDY_ID:ST004093 ANALYSIS_ID:AN006785 PROJECT_ID:PR002571
VERSION             	1
CREATED_ON             	August 5, 2025, 2:01 pm
#PROJECT
PR:PROJECT_TITLE                 	Metabolite profiling of Ankrd55-/- TH17 Cells
PR:PROJECT_SUMMARY               	Genome-wide association studies (GWAS) have linked the locus encoding Ankyrin
PR:PROJECT_SUMMARY               	Repeat Domain 55 (ANKRD55) with numerous autoimmune diseases; however, its
PR:PROJECT_SUMMARY               	biological function and role in inflammation are unclear. Here, we demonstrate
PR:PROJECT_SUMMARY               	that Ankrd55-deficient mice are protected from T cell-mediated colitis but are
PR:PROJECT_SUMMARY               	more susceptible to Citrobacter rodentium infection. Mechanistically, Ankrd55
PR:PROJECT_SUMMARY               	deletion impairs CD4+ T cell proliferation and reduces effector cytokine
PR:PROJECT_SUMMARY               	production in T helper 17 (TH17) cells in a cell-intrinsic manner. ANKRD55 is
PR:PROJECT_SUMMARY               	associated with mitochondria, and its loss is associated with impaired
PR:PROJECT_SUMMARY               	mitochondrial respiration and activation of the LKB1 pathway. Consistently,
PR:PROJECT_SUMMARY               	IL-17 production can be rescued by the deletion of LKB1 in Ankrd55-deficient T
PR:PROJECT_SUMMARY               	cells. Altogether, our study implicates the protein ANKRD55 as a functional
PR:PROJECT_SUMMARY               	modulator of T cell metabolism that directly impacts TH17 responses,
PR:PROJECT_SUMMARY               	highlighting it as a potential target across multiple autoimmune diseases.
PR:INSTITUTE                     	Broad Institute of MIT and Harvard
PR:LAST_NAME                     	Xavier
PR:FIRST_NAME                    	Ramnik
PR:ADDRESS                       	415 Main Street
PR:EMAIL                         	rxavier@broadinstitute.org
PR:PHONE                         	+1 (617) 714-7379
PR:PROJECT_COMMENTS              	Jinjin Xu, Lingjia Kong, Elizabeth A. Creasey, Sneha Rath, Lei Deng, Julian
PR:PROJECT_COMMENTS              	Avila-Pacheco, Chenhao Li, Blayne A. Oliver, Tyler Dao, Angela R. Shih, Mark J.
PR:PROJECT_COMMENTS              	Daly, Alex K. Shalek, Clary B. Clish, Daniel B. Graham, Jacques Deguine
#STUDY
ST:STUDY_TITLE                   	Autoimmune Disease Risk Gene ANKRD55 Promotes T Cell Proliferation and Th17 2
ST:STUDY_TITLE                   	Effector Function Through Metabolic Modulation
ST:STUDY_TYPE                    	TH17 Cell cultures
ST:STUDY_SUMMARY                 	ANKRD55, a gene linked to various autoimmune diseases through Genome-wide
ST:STUDY_SUMMARY                 	association studies (GWAS), plays a critical role in T cell function and
ST:STUDY_SUMMARY                 	inflammation. This study shows that Ankrd55-deficient mice are protected from T
ST:STUDY_SUMMARY                 	cell-mediated colitis but more vulnerable to Citrobacter rodentium infection,
ST:STUDY_SUMMARY                 	due to impaired CD4⁺ T cell proliferation and reduced TH17 cytokine
ST:STUDY_SUMMARY                 	production. To define the alterations in cellular metabolism caused by a
ST:STUDY_SUMMARY                 	Ankrd55-/- mutation in TH17 cells, we applied four metabolic profiling methods
ST:STUDY_SUMMARY                 	to in vitro differentiated TH17 cells. We found nucleotides were elevated in
ST:STUDY_SUMMARY                 	Ankrd55-/- compared to WT TH17 cells, with one-carbon units’ provider-folate
ST:STUDY_SUMMARY                 	(10-formyl-THF) being enriched. This was in accordance with the upregulated
ST:STUDY_SUMMARY                 	expression of enzymes in the SGOC pathway observed in Ankrd55-/- cells.
ST:INSTITUTE                     	Broad Institute of MIT and Harvard
ST:LAST_NAME                     	Xavier
ST:FIRST_NAME                    	Ramnik
ST:ADDRESS                       	415 Main Street
ST:EMAIL                         	rxavier@broadinstitute.org
ST:PHONE                         	+1 (617) 714-7379
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:CELL_STRAIN_DETAILS           	Naïve CD4+ T cells from either wild-type or Ankrd55-deficient C57BL/6 animals
#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           	-	PREFA01	Sample source:T-cells | Genotype:NA | Treatment:QC-drift_correction	RAW_FILE_NAME(C8-POS)=B0000c_XAV_JinJin_TH17_CP-PREFA01; RAW_FILE_NAME(HILIC-POS)=B0000k_XAV_Jinjin_TH17_HP-PREFA01; RAW_FILE_NAME(C18-NEG)=B0000g_XAV_Jinjin_TH17_CN-PREFA01; RAW_FILE_NAME(HILIC-NEG)=B0000e_XAV_Jinjin_TH17_HN-PREFA01
SUBJECT_SAMPLE_FACTORS           	-	PREFB01	Sample source:T-cells | Genotype:NA | Treatment:QC-pooled_ref	RAW_FILE_NAME(C8-POS)=B0000d_XAV_JinJin_TH17_CP-PREFB01; RAW_FILE_NAME(HILIC-POS)=B0000l_XAV_Jinjin_TH17_HP-PREFB01; RAW_FILE_NAME(C18-NEG)=B0000h_XAV_Jinjin_TH17_CN-PREFB01; RAW_FILE_NAME(HILIC-NEG)=B0000f_XAV_Jinjin_TH17_HN-PREFB01
SUBJECT_SAMPLE_FACTORS           	-	7	Sample source:T-cells | Genotype:WT | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	RAW_FILE_NAME(C8-POS)=B00001_XAV_JinJin_TH17_CP-Th17n_WT-1; RAW_FILE_NAME(HILIC-POS)=B0001_XAV_Jinjin_TH17_HP-7; RAW_FILE_NAME(C18-NEG)=B0001_XAV_Jinjin_TH17_CN-7; RAW_FILE_NAME(HILIC-NEG)=B0001_XAV_Jinjin_TH17_HN-7
SUBJECT_SAMPLE_FACTORS           	-	2	Sample source:T-cells | Genotype:WT | Treatment:No added cytokines	RAW_FILE_NAME(C8-POS)=B00002_XAV_JinJin_TH17_CP-Th0_WT-2; RAW_FILE_NAME(HILIC-POS)=B0002_XAV_Jinjin_TH17_HP-2; RAW_FILE_NAME(C18-NEG)=B0002_XAV_Jinjin_TH17_CN-2; RAW_FILE_NAME(HILIC-NEG)=B0002_XAV_Jinjin_TH17_HN-2
SUBJECT_SAMPLE_FACTORS           	-	3	Sample source:T-cells | Genotype:WT | Treatment:No added cytokines	RAW_FILE_NAME(C8-POS)=B00003_XAV_JinJin_TH17_CP-Th0_WT-3; RAW_FILE_NAME(HILIC-POS)=B0003_XAV_Jinjin_TH17_HP-3; RAW_FILE_NAME(C18-NEG)=B0003_XAV_Jinjin_TH17_CN-3; RAW_FILE_NAME(HILIC-NEG)=B0003_XAV_Jinjin_TH17_HN-3
SUBJECT_SAMPLE_FACTORS           	-	5	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:No added cytokines	RAW_FILE_NAME(C8-POS)=B00004_XAV_JinJin_TH17_CP-Th0_KO-2; RAW_FILE_NAME(HILIC-POS)=B0004_XAV_Jinjin_TH17_HP-5; RAW_FILE_NAME(C18-NEG)=B0004_XAV_Jinjin_TH17_CN-5; RAW_FILE_NAME(HILIC-NEG)=B0004_XAV_Jinjin_TH17_HN-5
SUBJECT_SAMPLE_FACTORS           	-	9	Sample source:T-cells | Genotype:WT | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	RAW_FILE_NAME(C8-POS)=B00005_XAV_JinJin_TH17_CP-Th17n_WT-3; RAW_FILE_NAME(HILIC-POS)=B0005_XAV_Jinjin_TH17_HP-9; RAW_FILE_NAME(C18-NEG)=B0005_XAV_Jinjin_TH17_CN-9; RAW_FILE_NAME(HILIC-NEG)=B0005_XAV_Jinjin_TH17_HN-9
SUBJECT_SAMPLE_FACTORS           	-	8	Sample source:T-cells | Genotype:WT | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	RAW_FILE_NAME(C8-POS)=B00006_XAV_JinJin_TH17_CP-Th17n_WT-2; RAW_FILE_NAME(HILIC-POS)=B0006_XAV_Jinjin_TH17_HP-8; RAW_FILE_NAME(C18-NEG)=B0006_XAV_Jinjin_TH17_CN-8; RAW_FILE_NAME(HILIC-NEG)=B0006_XAV_Jinjin_TH17_HN-8
SUBJECT_SAMPLE_FACTORS           	-	4	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:No added cytokines	RAW_FILE_NAME(C8-POS)=B00007_XAV_JinJin_TH17_CP-Th0_KO-1; RAW_FILE_NAME(HILIC-POS)=B0007_XAV_Jinjin_TH17_HP-4; RAW_FILE_NAME(C18-NEG)=B0007_XAV_Jinjin_TH17_CN-4; RAW_FILE_NAME(HILIC-NEG)=B0007_XAV_Jinjin_TH17_HN-4
SUBJECT_SAMPLE_FACTORS           	-	6	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:No added cytokines	RAW_FILE_NAME(C8-POS)=B00008_XAV_JinJin_TH17_CP-Th0_KO-3; RAW_FILE_NAME(HILIC-POS)=B0008_XAV_Jinjin_TH17_HP-6; RAW_FILE_NAME(C18-NEG)=B0008_XAV_Jinjin_TH17_CN-6; RAW_FILE_NAME(HILIC-NEG)=B0008_XAV_Jinjin_TH17_HN-6
SUBJECT_SAMPLE_FACTORS           	-	12	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	RAW_FILE_NAME(C8-POS)=B00009_XAV_JinJin_TH17_CP-Th17n_KO-3; RAW_FILE_NAME(HILIC-POS)=B0009_XAV_Jinjin_TH17_HP-12; RAW_FILE_NAME(C18-NEG)=B0009_XAV_Jinjin_TH17_CN-12; RAW_FILE_NAME(HILIC-NEG)=B0009_XAV_Jinjin_TH17_HN-12
SUBJECT_SAMPLE_FACTORS           	-	1	Sample source:T-cells | Genotype:WT | Treatment:No added cytokines	RAW_FILE_NAME(C8-POS)=B00010_XAV_JinJin_TH17_CP-Th0_WT-1; RAW_FILE_NAME(HILIC-POS)=B0010_XAV_Jinjin_TH17_HP-1; RAW_FILE_NAME(C18-NEG)=B0010_XAV_Jinjin_TH17_CN-1; RAW_FILE_NAME(HILIC-NEG)=B0010_XAV_Jinjin_TH17_HN-1
SUBJECT_SAMPLE_FACTORS           	-	10	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	RAW_FILE_NAME(C8-POS)=B00011_XAV_JinJin_TH17_CP-Th17n_KO-1; RAW_FILE_NAME(HILIC-POS)=B0011_XAV_Jinjin_TH17_HP-10; RAW_FILE_NAME(C18-NEG)=B0011_XAV_Jinjin_TH17_CN-10; RAW_FILE_NAME(HILIC-NEG)=B0011_XAV_Jinjin_TH17_HN-10
SUBJECT_SAMPLE_FACTORS           	-	11	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	RAW_FILE_NAME(C8-POS)=B00012_XAV_JinJin_TH17_CP-Th17n_KO-2; RAW_FILE_NAME(HILIC-POS)=B0012_XAV_Jinjin_TH17_HP-11; RAW_FILE_NAME(C18-NEG)=B0012_XAV_Jinjin_TH17_CN-11; RAW_FILE_NAME(HILIC-NEG)=B0012_XAV_Jinjin_TH17_HN-11
SUBJECT_SAMPLE_FACTORS           	-	PREFA02	Sample source:T-cells | Genotype:NA | Treatment:QC-drift_correction	RAW_FILE_NAME(C8-POS)=B0012a_XAV_JinJin_TH17_CP-PREFA02; RAW_FILE_NAME(HILIC-POS)=B0012a_XAV_Jinjin_TH17_HP-PREFA02; RAW_FILE_NAME(C18-NEG)=B0012a_XAV_Jinjin_TH17_CN-PREFA02; RAW_FILE_NAME(HILIC-NEG)=B0012a_XAV_Jinjin_TH17_HN-PREFA02
SUBJECT_SAMPLE_FACTORS           	-	PREFB02	Sample source:T-cells | Genotype:NA | Treatment:QC-pooled_ref	RAW_FILE_NAME(C8-POS)=B0012b_XAV_JinJin_TH17_CP-PREFB02; RAW_FILE_NAME(HILIC-POS)=B0012b_XAV_Jinjin_TH17_HP-PREFB02; RAW_FILE_NAME(C18-NEG)=B0012b_XAV_Jinjin_TH17_CN-PREFB02; RAW_FILE_NAME(HILIC-NEG)=B0012b_XAV_Jinjin_TH17_HN-PREFB02
#COLLECTION
CO:COLLECTION_SUMMARY            	Three biological replicates were generated for each WT and Ankrd55-/- cell
CO:COLLECTION_SUMMARY            	lines. Naïve CD4+ T cells from either wild-type or Ankrd55-/- C57BL/6 animals
CO:COLLECTION_SUMMARY            	were isolated by using EasySep Mouse Naïve CD4+ T Cell isolation Kit (STEMCELL
CO:COLLECTION_SUMMARY            	technologies, Vancouver Canada) and 1 million/mL of cells was cultured in 2
CO:COLLECTION_SUMMARY            	μg/mL anti-CD3/anti-CD28-coated 96-well plates under different cytokine
CO:COLLECTION_SUMMARY            	conditions for 3 days. Cells were harvested on day 3 and pellets were
CO:COLLECTION_SUMMARY            	snap-frozen and preserved at -80C until extraction. A polar extract was prepared
CO:COLLECTION_SUMMARY            	using 1,800,000 cells extracted in 80% methanol, while a lipid extract was
CO:COLLECTION_SUMMARY            	obtained from 600,000 cells using 100% isopropanol. The cleared extracts were
CO:COLLECTION_SUMMARY            	analyzed with a combination of four previously described liquid chromatography
CO:COLLECTION_SUMMARY            	high-resolution mass spectrometry (LC-MS) methods. The polar methanol extract
CO:COLLECTION_SUMMARY            	was analyzed using the HILIC-pos (10 μL), HILIC-neg (30 μL), and C18-neg (30
CO:COLLECTION_SUMMARY            	μL) methods, which target polar metabolites and those of intermediate polarity.
CO:COLLECTION_SUMMARY            	In contrast, the isopropanol extract was profiled using the C8-pos lipid
CO:COLLECTION_SUMMARY            	profiling method.
CO:SAMPLE_TYPE                   	T-cells
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	Cells were treated with different cytokine conditions for three days: Th0: no
TR:TREATMENT_SUMMARY             	added cytokines. Th17n: 25ng/mL IL-6 and 2ng/mL TGF-b1.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	LC–MS samples were profiled in each LC-MS method as follows: - HILIC-pos:
SP:SAMPLEPREP_SUMMARY            	Extracts (10 μL) were extracted with the addition of nine volumes of
SP:SAMPLEPREP_SUMMARY            	74.9:24.9:0.2 v/v/v acetonitrile/methanol/formic acid containing stable
SP:SAMPLEPREP_SUMMARY            	isotope-labeled internal standards (valine-d8, Isotec; and phenylalanine-d8,
SP:SAMPLEPREP_SUMMARY            	Cambridge Isotope Laboratories). The samples were centrifuged (10 min, 9,000g,
SP:SAMPLEPREP_SUMMARY            	4°C), and the supernatants (10 μL) injected directly onto column. - C8-pos:
SP:SAMPLEPREP_SUMMARY            	Extracts (10 μL) using 190 μL isopropanol containing
SP:SAMPLEPREP_SUMMARY            	1-dodecanoyl-2-tridecanoyl-sn-glycero-3-phosphocholine as an internal standard
SP:SAMPLEPREP_SUMMARY            	(Avanti Polar Lipids; Alabaster, AL). After centrifugation (10 min, 9,000g,
SP:SAMPLEPREP_SUMMARY            	ambient temperature), supernatants (2 μL) were injected directly onto column. -
SP:SAMPLEPREP_SUMMARY            	C18-neg: Extracts (30 μL) were extracted using 90 μl methanol containing
SP:SAMPLEPREP_SUMMARY            	15R-15-methyl Prostaglandin A2,15R-15-methyl Prostaglandin F2α, 15S-15-methyl
SP:SAMPLEPREP_SUMMARY            	Prostaglandin D2, 15S-15-methyl Prostaglandin E1, and 15S-15-methyl
SP:SAMPLEPREP_SUMMARY            	Prostaglandin E2 as internal standards (Cayman Chemical Co.) and centrifuged (10
SP:SAMPLEPREP_SUMMARY            	min, 9,000g, 4°C). The supernatants (10 μL) were injected onto column. -
SP:SAMPLEPREP_SUMMARY            	HILIC-neg: Extracts (30 μL) were extracted with the addition of four volumes of
SP:SAMPLEPREP_SUMMARY            	80% methanol containing inosine-15N4, thymine-d4 and glycocholate-d4 internal
SP:SAMPLEPREP_SUMMARY            	standards (Cambridge Isotope Laboratories). The samples were centrifuged (10
SP:SAMPLEPREP_SUMMARY            	min, 9,000g, 4°C) and the supernatants 10 μL) were injected directly onto
SP:SAMPLEPREP_SUMMARY            	column.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Shimadzu Nexera X2
CH:COLUMN_NAME                   	Phenomenex Luna NH2 (150 x 2 mm, 5 um)
CH:SOLVENT_A                     	100% water; 20 mM ammonium acetate; 20 mM ammonium hydroxide
CH:SOLVENT_B                     	75% acetonitrile/25% methanol; 10 mM ammonium hydroxide
CH:FLOW_GRADIENT                 	Elution began with 10% mobile phase A and 90% mobile phase B, followed by a
CH:FLOW_GRADIENT                 	linear gradient to 100% mobile phase A over 10 minutes. The gradient was then
CH:FLOW_GRADIENT                 	returned to 10% mobile phase A over 2 minutes and held for 13 minutes to
CH:FLOW_GRADIENT                 	equilibrate the column. MS acquisition stopped at 16 minutes.
CH:FLOW_RATE                     	400 µL/min
CH:COLUMN_TEMPERATURE            	30°C
CH:INTERNAL_STANDARD             	Inosine-15N4,Thymine-d4, Glycocholate-d4, CIL NLM-4264-0.01, DLM-2742-0,
CH:INTERNAL_STANDARD             	DLM-1089-1 respectively
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Plus Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Raw data were processed using TraceFinder 3.3 software (Thermo Fisher
MS:MS_COMMENTS                   	Scientific; Waltham, MA) and Progenesis QI (Nonlinear Dynamics; Newcastle upon
MS:MS_COMMENTS                   	Tyne, UK). Metabolite identities were confirmed using authentic reference
MS:MS_COMMENTS                   	standards or reference samples.
MS:MS_RESULTS_FILE               	ST004093_AN006785_Results.txt	UNITS:Abudances	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Abudances
MS_METABOLITE_DATA_START
Samples	PREFA01	PREFB01	7	2	3	5	9	8	4	6	12	1	10	11	PREFA02	PREFB02
Factors	Sample source:T-cells | Genotype:NA | Treatment:QC-drift_correction	Sample source:T-cells | Genotype:NA | Treatment:QC-pooled_ref	Sample source:T-cells | Genotype:WT | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	Sample source:T-cells | Genotype:WT | Treatment:No added cytokines	Sample source:T-cells | Genotype:WT | Treatment:No added cytokines	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:No added cytokines	Sample source:T-cells | Genotype:WT | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	Sample source:T-cells | Genotype:WT | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:No added cytokines	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:No added cytokines	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	Sample source:T-cells | Genotype:WT | Treatment:No added cytokines	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	Sample source:T-cells | Genotype:ankrd55 ko | Treatment:25ng/mL IL-6 and 2ng/mL TGF-b1	Sample source:T-cells | Genotype:NA | Treatment:QC-drift_correction	Sample source:T-cells | Genotype:NA | Treatment:QC-pooled_ref
Hippuric acid	816652	789526	475819	597083	573563	628525	543939	583118	864314	749018	993238	708684	882338	715556	801826	789526
Homovanillic acid	206641	190201	179339	152134	151197	120221	185327	221554	187127	108126	240321	90833	241656	200139	243247	190201
Dihexose	807751	710651	702246	184398	1205909	819096	541730	586384	308622				430332	248865	731556	710651
Pentose 5-phosphate	1986054	2204283	2609459	286464	257224	287077	3177321	3019252	402861	333057	6890926	291476	4472831	5188956	2185489	2204283
Hexose monophosphate	1915840	1851580	1086001	1678709	1449051	1264503	1188635	1350092	1993185	1987620	1859098	2174539	2373514	2062608	1786182	1851580
Hexose diphosphate	489151	491933	629111	192633	165220	296734	466202	620851	201448	260981	1228952	195478	560218	846899	493182	491933
Glyceric acid	1832419	1828598	1817000	1393769	1830848	1939648	1821932	1588520	1429130	1341706	1559465	1577936	1490774	1481125	1870322	1828598
Erythronic and or Threonic acid	40886360	38081780	39232130	32163225	27882186	30080325	39533557	48386210	38083004	26150356	46330227	28892153	47348792	40318595	39261763	38081780
Ribitol	6125184	6335330	3024205	8070832	7837196	8384540	2500352	2872824	10516649	6897825	4134209	7807814	3907686	3006742	6613768	6335330
Sorbitol	110732669	112273186	67264756	126360791	110504145	137600300	56273198	67587483	173363729	114898207	106909942	115600561	79554291	65969021	113521125	112273186
2-Aminobutyric acid	353757	391734	339860	357486	281016	400967	252075	234335	450438	334636	556139	369260	335648	268868	394840	391734
Oxalic acid	50174862	44264478	49967465	53012765	48260847	45784111	48942865	45667534	41664792	41969905	49300981	45473005	45259831	49671794	44740388	44264478
Adipic and or Methylglutaric acid	1504296	1549664	1289897	1334987	1618498	1317613	1375915	1240790	1434920	1188129	1115339	1847525	1424596	1125775	1597451	1549664
Aminoadipic acid	83111	92320		172058	134819	146039			192316	127893		160153	76417		163956	92320
Suberic acid	693581	699183	473659	339602	691867	695762	605171	641488	516050	558292	651653	648303	625730	646097	702887	699183
Hydroxybutyric and or Hydroxyisobutyric acid	1605797	1900399	1622343	2664489	2160742	2609088	1816669	2036668	3180675	2008912	1687444	1955905	1624190	1446913	1978213	1900399
2-Hydroxyvaleric and or 2-Hydroxyisovaleric acid	981186	1111553	851980	1266364	1244828	1388909	747099	921973	1624765	1008339	916130	1114428	955799	835468	1152857	1111553
2-Hydroxycaproic and or 2-Hydroxyisocaproic acid	686034	777470	1097069	1025518	852099	803272	637990	924116	1067215	580612	578358	633773	517752	579480	763785	777470
2-Hydroxyglutaric acid	9141518	9061036	9618036	6436517	7939863	9795374	8084708	7806710	11830134	8029166	9695826	11356931	8346885	7084398	9016070	9061036
NAD+	184390	189309		559184	448654	279012			502451	254627		317033			187419	189309
Adenine	910712	944191	508052	1215252	1438589	1091032	465373	663889	2624432	778637	500885	969246	451373	385319	930247	944191
Hypoxanthine	11192053	10761601	14825589				20901486	22305204			29516713	153075	20745512	23678879	10537345	10761601
Guanine	217574	264639	400808				574822	606847			814638		603746	669435	194910	264639
ADP	11926662	11218954	5938477	16194350	14628050	15236109	5602948	6627118	21416681	10961986	6555373	13111880	5450415	4910515	11143743	11218954
GDP	1363960	1380466	1195621	1491664	1369012	1323889	1126110	1295444	1966964	937452	1302498	1360178	1207836	1081416	1376426	1380466
AMP	16881304	16435566	14046741	16006311	14979303	15625437	13542458	17968777	21482796	11310303	14151509	14809774	20094203	15153898	16582915	16435566
IMP	738931	652021	551221	651407	709253	516553	378415	668888	941340	464267	574141	752847	880036	650281	631832	652021
GMP	2966245	2915206	2061351	2470688	2191190	2124784	2454448	3058179	3349441	1680530	3274957	2334778	4114520	3642464	2813192	2915206
Adenylsuccinic acid	312824	328354	225072	422682	316371	307808	172403	275549	518174	172468	219218	330003	355314	189810	312607	328354
ATP	5279033	5112626	1474727	8415191	6976034	8348088	1268535	1365517	11058661	6651310	1299794	7019379	792108	657439	4849043	5112626
Inosine	10767678	10728064	14019461				24245189	21307220			32783449		22272979	23893929	10597383	10728064
Guanosine	812160	837999	907913				1589659	1397108			3065963		1652475	2344504	823358	837999
Xanthine	2782568	2756238	1844679	3615875	3230020	3769097	1532941	2219645	4761003	2910914	1637294	2984982	1924204	1246299	2658928	2756238
Uric acid	1144925	1327232	1051541	1732104	1243584	1370110	851926	1211181	1635017	1298121	1250635	1232870	1164023	1025996	1329283	1327232
dTMP	107752	153059	70996	131536	104384	146436	37131	88041	198923	111119	203319	87439	184962	178775	155706	153059
CDP	1188152	1165880	1188859	1051531	1002439	1183961	1000174	1246131	1544466	988724	1395303	978818	938212	921864	1210328	1165880
UDP	3413459	3454633	3565472	3098625	2635414	3189326	2831356	3634873	4201903	2884361	4366469	3025356	3066144	2838091	3589657	3454633
UDP-galactose or UDP-glucose	2306374	2170776	2311825	1970826	1559867	1972416	1733019	2087373	2676675	2026510	3175967	1863784	1712279	1936555	2034455	2170776
CMP	1584932	1603937	1423284	1084286	978899	1288161	1515545	1776773	1573276	986314	2459086	1000272	2264388	2178023	1663931	1603937
UMP	7300353	7563506	7077308	5169992	4798624	5140360	6675169	8334693	7283187	4690116	11118633	5350231	10797600	9499155	7694650	7563506
Cytidine	704599	741990	862869	166669	219491	247081	1129498	1132488	445684	170693	1910352	145655	1026400	1424723	714066	741990
Uridine	36856775	35281322	38095202	5065230	5110158	6435340	50326659	46867852	8839747	5249594	124553535	5792834	57243441	79889449	35812991	35281322
Pseudouridine	484663	453020	271434	563906	386756	593909	360993	414863	789518	418071	368611	333307	293557	335236	441204	453020
Uracil	14631150	14382396	16969144	2635137	2708273	3189328	21630367	21712113	4238055	2885960	44489918	2761910	24705537	28204288	14397714	14382396
Aspartic acid	48538573	47924890	44422832	46387680	37391528	45420343	33036032	38441976	50385501	53159874	66349744	42969636	34736479	34866984	49643890	47924890
Glutamic acid	161619795	161216203	182805342	122971314	96416792	134249870	137417907	168141079	152649253	143271726	291348286	124431883	163186922	147939030	158366273	161216203
N-Acetylglutamic acid	803462	756053	789862	460132	463892	591076	765839	814044	607364	552413	1078506	563577	982100	895198	751579	756053
Pantothenic acid	9607860	8059071	6233350	7413321	7097398	7268331	4792145	6196009	9701400	7566330	9947896	8428287	8489733	6294155	8125532	8059071
Glutathione	111100115	109809249	112118533	92585652	84481204	81690594	98399809	103146965	100518388	86616250	168808512	97992054	129130031	121502741	110223740	109809249
Oxidized glutathione	107189	132411	107120	46095	41931	166238	179954	146724	158832	51392	201391	41831	133940	104591	129295	132411
Ketocaproic and or Ketoisocaproic acid	917464	885576		1487983	959463	1452627			1874381	1278733		1123361			940367	885576
sn-Glycero-3-phosphate	306341	276661	175296	319485	284446	304520	205197	224069	300283	233545	184750	265915	275346	194967	261771	276661
Lactic acid	900331270	1041127450	757166045	1190102532	1111126015	1143095131	817446726	906115645	1316943169	1080827966	855395460	1106439443	1070617507	854753604	1050411359	1041127450
3-Phosphoglyceric acid	678699	605067	354935	603029	647725	777734	387823	489834	815451	842419	630850	702542	415978	462569	593412	605067
2-Phosphoglyceric acid	129058	93588		104705	99206	144227	42743	90033	166269	144373	100511	140513	65805	59189	80195	93588
Fumaric and or Maleic acid	173691537	173740651	105390705	204605283	171852241	203576143	96452521	108400161	260712171	199736970	156433880	195503750	139281114	119863430	182204016	173740651
Succinic acid	69055024	67933070	59251531	62665036	67269531	65801694	58340874	63414739	71110153	68548347	64993219	72717683	71407483	62941222	67567302	67933070
Malic acid	377520579	365875904	224956353	436553851	357827476	432351861	206741342	228001513	541193470	426925561	325542235	402138242	289361387	251593571	377139053	365875904
cis-Aconitic acid	2582731	2186405	2706617	2131441	1845309	2036354	2352723	2627405	2539996	1591993	2522654	1580487	1676020	2018459	2211941	2186405
Citric and or Isocitric acid	51873741	47056642	52740360	45421827	40654127	45864702	47843642	52601883	56331610	39976984	52800217	35361044	34753618	43310436	46590330	47056642
Myo-inositol	17472544	17206004	11591779	17901519	15854027	18882113	11237935	13483371	23005296	14587988	17076627	14801708	14069188	12839475	17113778	17206004
Malondialdehyde	3540771	3705165	2704090	3453297	3410291	3457672	3016177	3217140	4395940	3366717	2732638	3292541	3491913	2873799	3119022	3705165
Glucosan	3146854	3097566	2870667	2930709	2898515	3112242	2642295	2750931	3416070	2973417	3141998	2727527	2887704	2818166	3049332	3097566
Taurdeoxycholic and or Taurochenodeoxycholic acid	263472	260640	165577	274441	243868	338833	80160	173434	393915	362962	211849	155406	272939	212575	277623	260640
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Study_Compound_ID	MZ	RT	HMDB_ID
Hippuric acid	TF0086	178.05097000000001	4.3899999999999997	HMDB0000714
Homovanillic acid	TF0088	181.05063000000001	4.41	HMDB0000118
Dihexose	TF0141	341.10892999999999	2.29	HMDB0000258
Pentose 5-phosphate	TF0122	229.01187999999999	7.39	HMDB0001548
Hexose monophosphate	TF0085	259.02244000000002	7.34	HMDB0000124
Hexose diphosphate	TF0084	338.98876999999999	10.43	HMDB0001058
Glyceric acid	TF0077	105.01933	4.18	HMDB0000139
Erythronic and or Threonic acid	TF0064	135.0299	4.18	HMDB0000613
Ribitol	TF0034	151.06120000000001	1.71	HMDB0000508
Sorbitol	TF0137	181.07176000000001	1.91	HMDB0000247
2-Aminobutyric acid	TF0005	102.05605	2.96	HMDB0000650
Oxalic acid	TF0119	88.988029999999995	7.57	HMDB0002329
Adipic and or Methylglutaric acid	TF0033	145.05063000000001	6.86	HMDB0000448
Aminoadipic acid	TF0004	160.06153	5.18	HMDB0000510
Suberic acid	TF0138	173.08193	6.87	HMDB0000893
Hydroxybutyric and or Hydroxyisobutyric acid	TF0037	103.04007	3.72	HMDB0000008
2-Hydroxyvaleric and or 2-Hydroxyisovaleric acid	TF0007	117.05571999999999	3.57	HMDB0000407
2-Hydroxycaproic and or 2-Hydroxyisocaproic acid	TF0008	131.07137	3.56	HMDB0000317
2-Hydroxyglutaric acid	TF0009	147.0299	6.91	HMDB0000694
NAD+	TF0112	662.10239000000001	4.8499999999999996	HMDB0000902
Adenine	TF0031	134.04722000000001	2.2799999999999998	HMDB0000034
Hypoxanthine	TF0089	135.03122999999999	3.36	HMDB0000157
Guanine	TF0082	150.04212999999999	2.87	HMDB0000132
ADP	TF0035	426.02213999999998	10.199999999999999	HMDB0001341
GDP	TF0071	442.01704999999998	11.17	HMDB0001201
AMP	TF0041	346.05581000000001	7.73	HMDB0000045
IMP	TF0090	347.03982000000002	9	HMDB0000175
GMP	TF0081	362.05072000000001	8.6999999999999993	HMDB0001397
Adenylsuccinic acid	TF0032	462.06677000000002	10.68	HMDB0000536
ATP	TF0044	505.98847000000001	12.46	HMDB0000538
Inosine	TF0095	267.07348999999999	3.4	HMDB0000195
Guanosine	TF0083	282.08438999999998	3	HMDB0000133
Xanthine	TF0158	151.02615	5.04	HMDB0000292
Uric acid	TF0155	167.02106000000001	5.27	HMDB0000289
dTMP	TF0062	321.04933	7.42	HMDB0001227
CDP	TF0049	402.01089999999999	9.69	HMDB0001546
UDP	TF0150	402.99491999999998	10.08	HMDB0000295
UDP-galactose or UDP-glucose	TF0151	565.04773999999998	7.18	HMDB0000286
CMP	TF0054	322.04457000000002	7.47	HMDB0000095
UMP	TF0153	323.02859000000001	7.71	HMDB0000288
Cytidine	TF0059	242.07823999999999	2.0299999999999998	HMDB0000089
Uridine	TF0156	243.06226000000001	1.76	HMDB0000296
Pseudouridine	TF0128	243.06226000000001	2.34	HMDB0000767
Uracil	TF0154	111.02	1.54	HMDB0000300
Aspartic acid	TF0043	132.03022999999999	5.12	HMDB0000191
Glutamic acid	TF0074	146.04588000000001	5.09	HMDB0000148
N-Acetylglutamic acid	TF0111	188.05645000000001	6.71	HMDB0001138
Pantothenic acid	TF0121	218.10339999999999	3.85	HMDB0000210
Glutathione	TF0076	306.07652999999999	5.97	HMDB0000125
Oxidized glutathione	TF0075	611.14467999999999	8.0399999999999991	HMDB0003337
Ketocaproic and or Ketoisocaproic acid	TF0038	129.05572000000001	3.36	HMDB0000491
sn-Glycero-3-phosphate	TF0036	171.00640000000001	7.34	HMDB0000126
Lactic acid	TF0102	89.024420000000006	3.87	HMDB0000190
3-Phosphoglyceric acid	TF0019	184.98566	9.44	HMDB0000807
2-Phosphoglyceric acid	TF0011	184.98566	9.6	HMDB0000362
Fumaric and or Maleic acid	TF0070	115.00368	7.04	HMDB0000134
Succinic acid	TF0139	117.01933	6.98	HMDB0000254
Malic acid	TF0104	133.01425	7.04	HMDB0000156
cis-Aconitic acid	TF0030	173.00916000000001	9.2799999999999994	HMDB0000072
Citric and or Isocitric acid	TF0053	191.01973000000001	9.2799999999999994	HMDB0000094
Myo-inositol	TF0097	179.05610999999999	2.58	HMDB0000211
Malondialdehyde	TF0106	71.013850000000005	3.86	HMDB0006112
Glucosan	TF0042	161.04554999999999	6.84	HMDB0000640
Taurdeoxycholic and or Taurochenodeoxycholic acid	TF0144	498.28948000000003	3.49	HMDB0000896
METABOLITES_END
#END