#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