#METABOLOMICS WORKBENCH iuk41_20240328_131847 DATATRACK_ID:4750 STUDY_ID:ST003150 ANALYSIS_ID:AN005168 PROJECT_ID:PR001958
VERSION                          	1
CREATED_ON                       	08-28-2024
#PROJECT
PR:PROJECT_TITLE                 	Impact of early-life exposure to a potent aryl hydrocarbon receptor ligand on
PR:PROJECT_TITLE                 	gut microbiota and host glucose homeostasis in C57BL/6J male mice
PR:PROJECT_SUMMARY               	This study aimed to explore the association between early-life exposure to a
PR:PROJECT_SUMMARY               	potent aryl hydrocarbon receptor (AHR) agonist and persistent disruptions in the
PR:PROJECT_SUMMARY               	microbiota, leading to impaired metabolic homeostasis later in life. This study
PR:PROJECT_SUMMARY               	utilized metagenomics, NMR- and mass spectrometry-based metabolomics, and
PR:PROJECT_SUMMARY               	biochemical assays to analyze the gut microbiome composition and function, as
PR:PROJECT_SUMMARY               	well as the physiological and metabolic effects of early-life exposure to
PR:PROJECT_SUMMARY               	2,3,7,8-tetrachlorodibenzofuran (TCDF) in conventional, germ-free (GF), and
PR:PROJECT_SUMMARY               	Ahr-null mice. The impact of TCDF on Akkermansia muciniphila (A. muciniphila) in
PR:PROJECT_SUMMARY               	vitro was assessed using optical density (OD 600), flow cytometry,
PR:PROJECT_SUMMARY               	transcriptomics, and mass spectrometry-based metabolomics. TCDF-exposed mice
PR:PROJECT_SUMMARY               	exhibited disruption in the gut microbiome community structure and function,
PR:PROJECT_SUMMARY               	characterized by lower abundances of A. muciniphila, lower levels of cecal short
PR:PROJECT_SUMMARY               	chain fatty acids (SCFAs) and indole-3-lactic acid (ILA), and a reduction in gut
PR:PROJECT_SUMMARY               	hormones GLP-1 and PYY. Importantly, microbial and metabolic phenotypes
PR:PROJECT_SUMMARY               	associated with early-life POP exposure were transferable to GF recipients in
PR:PROJECT_SUMMARY               	the absence of POP carry-over. In addition, AHR-independent interactions between
PR:PROJECT_SUMMARY               	POPs and the microbiota were observed, significantly affected the growth,
PR:PROJECT_SUMMARY               	physiology, gene expression, and metabolic activity of A. muciniphila, resulting
PR:PROJECT_SUMMARY               	in suppressed activity along the ILA pathway.
PR:INSTITUTE                     	Pennsylvania State University
PR:DEPARTMENT                    	Department of Veterinary and Biomedical Sciences
PR:LAST_NAME                     	Koo
PR:FIRST_NAME                    	Imhoi
PR:ADDRESS                       	307B Life Science Building
PR:EMAIL                         	iuk41@psu.edu
PR:PHONE                         	+1 814-865-7803
PR:DOI                           	http://dx.doi.org/10.21228/M8MX5P
#STUDY
ST:STUDY_TITLE                   	Impact of early-life exposure to a potent aryl hydrocarbon receptor ligand on
ST:STUDY_TITLE                   	gut microbiota and host glucose homeostasis in C57BL/6J male mice (Part II)
ST:STUDY_SUMMARY                 	This study aimed to explore the association between early-life exposure to a
ST:STUDY_SUMMARY                 	potent aryl hydrocarbon receptor (AHR) agonist and persistent disruptions in the
ST:STUDY_SUMMARY                 	microbiota, leading to impaired metabolic homeostasis later in life. This study
ST:STUDY_SUMMARY                 	utilized metagenomics, NMR- and mass spectrometry-based metabolomics, and
ST:STUDY_SUMMARY                 	biochemical assays to analyze the gut microbiome composition and function, as
ST:STUDY_SUMMARY                 	well as the physiological and metabolic effects of early-life exposure to
ST:STUDY_SUMMARY                 	2,3,7,8-tetrachlorodibenzofuran (TCDF) in conventional, germ-free (GF), and
ST:STUDY_SUMMARY                 	Ahr-null mice. The impact of TCDF on Akkermansia muciniphila (A. muciniphila) in
ST:STUDY_SUMMARY                 	vitro was assessed using optical density (OD 600), flow cytometry,
ST:STUDY_SUMMARY                 	transcriptomics, and mass spectrometry-based metabolomics. TCDF-exposed mice
ST:STUDY_SUMMARY                 	exhibited disruption in the gut microbiome community structure and function,
ST:STUDY_SUMMARY                 	characterized by lower abundances of A. muciniphila, lower levels of cecal short
ST:STUDY_SUMMARY                 	chain fatty acids (SCFAs) and indole-3-lactic acid (ILA), and a reduction in gut
ST:STUDY_SUMMARY                 	hormones GLP-1 and PYY. Importantly, microbial and metabolic phenotypes
ST:STUDY_SUMMARY                 	associated with early-life POP exposure were transferable to GF recipients in
ST:STUDY_SUMMARY                 	the absence of POP carry-over. In addition, AHR-independent interactions between
ST:STUDY_SUMMARY                 	POPs and the microbiota were observed, significantly affected the growth,
ST:STUDY_SUMMARY                 	physiology, gene expression, and metabolic activity of A. muciniphila, resulting
ST:STUDY_SUMMARY                 	in suppressed activity along the ILA pathway.
ST:INSTITUTE                     	Pennsylvania State University
ST:DEPARTMENT                    	Department of Veterinary and Biomedical Sciences
ST:LAST_NAME                     	Koo
ST:FIRST_NAME                    	Imhoi
ST:ADDRESS                       	307B Life Science Building
ST:EMAIL                         	iuk41@psu.edu
ST:PHONE                         	+1 814-865-7803
ST:SUBMIT_DATE                   	2024-03-28
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:AGE_OR_AGE_RANGE              	4 weeks to 5 months
SU:GENDER                        	Male
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	YT200819_08	YT200819_08	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_08.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_09	YT200819_09	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_09.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_10	YT200819_10	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_10.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_105	YT200819_105	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_105.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_106	YT200819_106	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_106.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_107	YT200819_107	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_107.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_108	YT200819_108	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_108.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_109	YT200819_109	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_109.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_11	YT200819_11	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_11.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_110	YT200819_110	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_110.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_12	YT200819_12	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_12.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_13	YT200819_13	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT200819_13.CDF
SUBJECT_SAMPLE_FACTORS           	YT201026_14	YT201026_14	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT201026_14.CDF
SUBJECT_SAMPLE_FACTORS           	YT201026_15	YT201026_15	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT201026_15.CDF
SUBJECT_SAMPLE_FACTORS           	YT201026_16	YT201026_16	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT201026_16.CDF
SUBJECT_SAMPLE_FACTORS           	YT201026_17	YT201026_17	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT201026_17.CDF
SUBJECT_SAMPLE_FACTORS           	YT201026_18	YT201026_18	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT201026_18.CDF
SUBJECT_SAMPLE_FACTORS           	YT201026_19	YT201026_19	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT201026_19.CDF
SUBJECT_SAMPLE_FACTORS           	YT210427_16	YT210427_16	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT210427_16.CDF
SUBJECT_SAMPLE_FACTORS           	YT210427_17	YT210427_17	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT210427_17.CDF
SUBJECT_SAMPLE_FACTORS           	YT210427_18	YT210427_18	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT210427_18.CDF
SUBJECT_SAMPLE_FACTORS           	YT210427_19	YT210427_19	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT210427_19.CDF
SUBJECT_SAMPLE_FACTORS           	YT210427_20	YT210427_20	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT210427_20.CDF
SUBJECT_SAMPLE_FACTORS           	YT210427_21	YT210427_21	Sample source:liver | Treatment:TCDF	RAW_FILE_NAME=YT210427_21.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_02	YT200819_02	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_02.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_03	YT200819_03	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_03.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_04	YT200819_04	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_04.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_05	YT200819_05	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_05.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_06	YT200819_06	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_06.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_07	YT200819_07	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_07.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_101	YT200819_101	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_101.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_102	YT200819_102	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_102.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_103	YT200819_103	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_103.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_104	YT200819_104	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_104.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_21	YT200819_21	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_21.CDF
SUBJECT_SAMPLE_FACTORS           	YT200819_22	YT200819_22	Sample source:liver | Treatment:vehicle	RAW_FILE_NAME=YT200819_22.CDF
#COLLECTION
CO:COLLECTION_SUMMARY            	Liver tissue samples were collected immediately after sacrifice by carbon
CO:COLLECTION_SUMMARY            	dioxide asphyxiation and kept at -80°C.
CO:SAMPLE_TYPE                   	Liver
#TREATMENT
TR:TREATMENT_SUMMARY             	1. After feed training, mice were fed pills containing TCDF (dissolved in
TR:TREATMENT_SUMMARY             	acetone) or acetone alone as vehicle, and one pill uniformly contained 0.46 µg
TR:TREATMENT_SUMMARY             	TCDF (24 µg/kg as final dose). Mice were housed singly in an empty cage and
TR:TREATMENT_SUMMARY             	monitored to ensure the pill was consumed in the morning for 5 days. 2. TCDF (24
TR:TREATMENT_SUMMARY             	µg/kg) or corn oil as vehicle were administered to age-matched male GF and
TR:TREATMENT_SUMMARY             	Ahr-/- mice by oral gavage once daily for five days (n = 4 per group). 3.
TR:TREATMENT_SUMMARY             	Four-week-old male GF C57BL/6J mice were orally gavaged with 100 µL of cecal
TR:TREATMENT_SUMMARY             	suspension (100 mg in 1 mL sterile BHI CHV media) from vehicle or TCDF treated
TR:TREATMENT_SUMMARY             	mice in long-duration model. 4. A. muciniphila was administered to GF mice by
TR:TREATMENT_SUMMARY             	oral gavage at one dose 107 CFU/0.1 mL suspended in sterile BHI CHV media
TR:TREATMENT_SUMMARY             	containing an end concentration of glycerol (15% vol/vol).
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	For liver lipid quantification, 50 mg of liver tissue was homogenized in 1 mL
SP:SAMPLEPREP_SUMMARY            	pre-cooled chloroform/methanol mix [1:1 (v/v)], followed by adding 296 µL
SP:SAMPLEPREP_SUMMARY            	water. After centrifugation (10,000 g, 4°C) for 10 min, the lower phase was
SP:SAMPLEPREP_SUMMARY            	dried in a vacuum and reconstituted in 600 µL of deuterated chloroform
SP:SAMPLEPREP_SUMMARY            	containing 0.03% (v/v) tetramethylsilane (TMS).
SP:SAMPLEPREP_PROTOCOL_FILENAME  	PSU_Methyl_esterification_GCMS_032724.pdf
#CHROMATOGRAPHY
CH:INSTRUMENT_NAME               	Agilent 7890A
CH:COLUMN_NAME                   	Restek Rtx-5Sil MS (30m x 0.25mm, 0.25um)
CH:COLUMN_TEMPERATURE            	325
CH:FLOW_GRADIENT                 	N/A
CH:FLOW_RATE                     	0.5658 mL/min
CH:SOLVENT_A                     	N/A
CH:SOLVENT_B                     	N/A
CH:CHROMATOGRAPHY_TYPE           	GC
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:ANALYSIS_PROTOCOL_FILE        	PSU_tissue_POP_measurement_GCMS_032724.pdf
#MS
MS:INSTRUMENT_NAME               	Agilent 5975C
MS:INSTRUMENT_TYPE               	Single quadrupole
MS:MS_TYPE                       	EI
MS:MS_COMMENTS                   	For ionization, the EI source was used with the following settings: filament
MS:MS_COMMENTS                   	source temperature at 230 °C and electron ionization energy at 70 eV. Mass
MS:MS_COMMENTS                   	spectra were collected in a mass range of m/z 50 - 600 at a scan rate of 2
MS:MS_COMMENTS                   	spectra/s. Mass calibration was performed after every injection with internal
MS:MS_COMMENTS                   	reference masses m/z 68.9947, 263.9866 and 501.9706. Data acquisition was
MS:MS_COMMENTS                   	performed using the Agilent MassHunter Workstation GC-MS Data Acquisition v 10.0
MS:MS_COMMENTS                   	(Agilent Technologies, Waldbronn, Germany).
MS:ION_MODE                      	POSITIVE
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	peak area
MS_METABOLITE_DATA_START
Samples	YT200819_08	YT200819_09	YT200819_10	YT200819_105	YT200819_106	YT200819_107	YT200819_108	YT200819_109	YT200819_11	YT200819_110	YT200819_12	YT200819_13	YT201026_14	YT201026_15	YT201026_16	YT201026_17	YT201026_18	YT201026_19	YT210427_16	YT210427_17	YT210427_18	YT210427_19	YT210427_20	YT210427_21	YT200819_02	YT200819_03	YT200819_04	YT200819_05	YT200819_06	YT200819_07	YT200819_101	YT200819_102	YT200819_103	YT200819_104	YT200819_21	YT200819_22
Factors	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:TCDF	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	Sample source:liver | Treatment:vehicle	
C16:0	14.2997	9.5912	9.4064	9.4939	8.2378	7.3132	7.5037	8.5706	14.9862	7.9629	12.9385	14.9787													9.7144	9.1099	10.4442	9.0768	6.5022	10.1233	8.1102	7.8665	7.4679	8.3550	6.6211	7.2737
C16:1	0.6559	0.3938	0.4402	0.3723	0.2916	0.2882	0.3254	0.5366	0.5627	0.2917	0.5439	0.6311													0.3070	0.3969	0.2538	0.1036	0.1335	0.2739	0.3339	0.3000	0.2527	0.4038	0.1998	0.1772
C18:0	8.2175	5.8504	5.5207	6.0644	5.7429	4.7699	4.5690	4.6303	8.3825	4.8435	7.8593	8.8543													6.0176	4.5402	5.9858	5.7271	4.6424	6.0643	4.7258	4.4433	4.5735	4.8137	4.5229	4.6019
C18:1n9c	10.8908	4.7369	6.2051	4.2172	3.3333	2.8347	3.9901	3.5964	11.1569	3.0368	8.2821	11.2905													4.7652	4.1170	4.9420	3.3062	2.8534	4.2944	3.4888	3.4077	3.1321	3.1791	2.8636	2.7683
C18:1n9t	0.7333	0.3518	0.5577	0.4150	0.2485	0.2761	0.4549	0.3861	1.0831	0.2295	0.8812	1.1376													0.4620	0.3254	0.3545	0.2323	0.2291	0.4426	0.2884	0.3075	0.2626	0.3506	0.2438	0.2108
C18:2n6c	11.1053	6.1613	6.6936	6.1849	5.6005	4.1106	4.6859	4.2681	10.5579	4.7059	9.5875	12.3991													6.2373	4.6073	6.7776	5.3067	3.7296	5.7873	4.4571	4.4252	4.2226	4.6884	3.7811	4.2270
C20:0	0.3854	0.2303	0.2804	0.1927	0.4707	0.2703	0.2603	0.3087	0.3946	0.2827	0.2935	0.3906													0.2548	0.1087	0.2219	0.2171	0.1166	0.2417	0.2013	0.1493	0.1735	0.1960	0.1820	0.1614
C20:3n6	0.5539	0.2123	0.4216	0.4349	0.3234	0.3083	0.3928	0.3771	0.7459	0.3243	0.7592	0.9122													0.2933	0.1801	0.1716	0.1824	0.1827	0.2916	0.3632	0.2842	0.2881	0.3147	0.2897	0.2153
C20:4n6	2.8664	1.9419	1.9168	2.6879	2.5845	2.0239	2.4543	2.2046	3.3237	2.1269	3.3672	3.8548													2.7365	1.5030	2.0968	2.0073	1.4317	2.7529	2.0491	1.8976	1.9674	2.1455	1.9049	2.0179
C20:5n3	0.1979	0.1423	0.1791	0.2621	0.2345	0.2521	0.1546	0.1415	0.2309	0.2632	0.2039	0.2239													0.2468	0.1489	0.2177	0.1196	0.1241	0.1491	0.1729	0.2167	0.1804	0.1793	0.1851	0.1468
C22:0	0.5098	0.3609	0.4098	0.2461	0.4268	0.3607	0.2772	0.2891	0.4356	0.3072	0.3024	0.4460													0.4112	0.1407	0.2509	0.3241	0.1527	0.2796	0.2322	0.1961	0.1982	0.1930	0.1790	0.1854
C22:6n3	3.1799	2.5534	2.2200	3.0394	2.8728	2.1774	2.2782	2.0398	3.8884	2.4971	3.4638	4.1465													3.0171	1.7347	2.2159	2.2845	1.4519	2.3225	2.3347	2.1138	2.1201	2.2802	1.9843	2.0473
TCDF													0.2312	0.1546	0.0817	0.0745	0.1398	0.1000
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	pubchem_id	inchi_key	kegg_id	other_id	other_id_type	ri	ri_type	moverz_quant	
C16:0	985								
C16:1									
C18:0	5281								
C18:1n9c									
C18:1n9t									
C18:2n6c									
C20:0	10467								
C20:3n6									
C20:4n6									
C20:5n3									
C22:0	8215								
C22:6n3									
TCDF	39929								
METABOLITES_END
#END