#METABOLOMICS WORKBENCH araskind_20200609_095708 DATATRACK_ID:2050 STUDY_ID:ST001398 ANALYSIS_ID:AN002337 VERSION 1 CREATED_ON 12-15-2022 #PROJECT PR:PROJECT_TITLE Mechanism of Trichloroethylene (TCE) toxicity in placenta PR:PROJECT_SUMMARY This study will provide important information on the mechanism of TCE toxicity PR:PROJECT_SUMMARY in the placenta PR:INSTITUTE University of Michigan PR:DEPARTMENT Environmental Health PR:LABORATORY Loch-Caruso Lab PR:LAST_NAME Loch-Caruso PR:FIRST_NAME Rita PR:ADDRESS Ann Arbor, MI PR:EMAIL rlc@umich.edu PR:PHONE 734-936-1256 PR:DOI http://dx.doi.org/10.21228/M8VM30 #STUDY ST:STUDY_TITLE Mechanism of Trichloroethylene (TCE) toxicity in the placenta ST:STUDY_TYPE MS analysis ST:STUDY_SUMMARY Cells in culture will be exposed to S-(1,2-dichlorovinyl)-L-cysteine (DCVC), a ST:STUDY_SUMMARY relevant metabolite of TCE. The first cell line used will be HTR-8/SVneo, ST:STUDY_SUMMARY originally derived from first trimester female human cytotrophoblast cells and ST:STUDY_SUMMARY immortalized with simian virus 40 large T antigen, this cell line models ST:STUDY_SUMMARY first-trimester placental extravillous trophoblasts in vitro. HTR-8/SVneo cells ST:STUDY_SUMMARY will be cultured for 24 hours followed by exposure to cell culture media ST:STUDY_SUMMARY (control) or 20 µM DCVC for 6 or 12 hours. Cells (cells frozen in cell culture ST:STUDY_SUMMARY dish) will then be collected (n=5) In the TCE aim of this study, we will ST:STUDY_SUMMARY utilizeone cell model that phenotypically represents an important placental cell ST:STUDY_SUMMARY population. ST:INSTITUTE University of Michigan ST:DEPARTMENT Biomedical Research Core Facilities ST:LABORATORY Metabolomics core ST:LAST_NAME Kachman ST:FIRST_NAME Maureen ST:ADDRESS Ann Arbor, MI ST:EMAIL mkachman@med.umich.edu ST:PHONE 734-232-0842 ST:SUBMIT_DATE 2020-06-08 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - S00034911 Hours of Exposure:12 hr | Treatment:20 uM DCVC Trial Number=1; Sample name=16_Elkin_N1_12hr_20 uM_DCVC_4-29-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034911-N SUBJECT_SAMPLE_FACTORS - S00034912 Hours of Exposure:12 hr | Treatment:20 uM DCVC Trial Number=2; Sample name=17_Elkin_N2_12hr_20 uM_DCVC_4-29-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034912-N SUBJECT_SAMPLE_FACTORS - S00034913 Hours of Exposure:12 hr | Treatment:20 uM DCVC Trial Number=3; Sample name=18_Elkin_N3_12hr_20 uM_DCVC_4-29-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034913-N SUBJECT_SAMPLE_FACTORS - S00034914 Hours of Exposure:12 hr | Treatment:20 uM DCVC Trial Number=4; Sample name=19_Elkin_N4_12hr_20 uM_DCVC_4-29-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034914-N SUBJECT_SAMPLE_FACTORS - S00034915 Hours of Exposure:12 hr | Treatment:20 uM DCVC Trial Number=5; Sample name=20_Elkin_N5_12hr_20 uM_DCVC_4-29-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034915-N SUBJECT_SAMPLE_FACTORS - S00034906 Hours of Exposure:12 hr | Treatment:No Treatment Trial Number=1; Sample name=11_Elkin_N1_12hr_NT_4-29-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034906-N SUBJECT_SAMPLE_FACTORS - S00034907 Hours of Exposure:12 hr | Treatment:No Treatment Trial Number=2; Sample name=12_Elkin_N2_12hr_NT_4-29-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034907-N SUBJECT_SAMPLE_FACTORS - S00034908 Hours of Exposure:12 hr | Treatment:No Treatment Trial Number=3; Sample name=13_Elkin_N3_12hr_NT_4-29-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034908-N SUBJECT_SAMPLE_FACTORS - S00034909 Hours of Exposure:12 hr | Treatment:No Treatment Trial Number=4; Sample name=14_Elkin_N4_12hr_NT_4-29-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034909-N SUBJECT_SAMPLE_FACTORS - S00034910 Hours of Exposure:12 hr | Treatment:No Treatment Trial Number=5; Sample name=15_Elkin_N5_12hr_NT_4-29-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034910-N SUBJECT_SAMPLE_FACTORS - S00034901 Hours of Exposure:6 hr | Treatment:20 uM DCVC Trial Number=1; Sample name=6_Elkin_N1_6hr_20 uM_DCVC_4-12-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034901-N SUBJECT_SAMPLE_FACTORS - S00034902 Hours of Exposure:6 hr | Treatment:20 uM DCVC Trial Number=2; Sample name=7_Elkin_N2_6hr_20 uM_DCVC_4-12-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034902-N SUBJECT_SAMPLE_FACTORS - S00034903 Hours of Exposure:6 hr | Treatment:20 uM DCVC Trial Number=3; Sample name=8_Elkin_N3_6hr_20 uM_DCVC_4-12-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034903-N SUBJECT_SAMPLE_FACTORS - S00034904 Hours of Exposure:6 hr | Treatment:20 uM DCVC Trial Number=4; Sample name=9_Elkin_N4_6hr_20 uM_DCVC_4-12-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034904-N SUBJECT_SAMPLE_FACTORS - S00034905 Hours of Exposure:6 hr | Treatment:20 uM DCVC Trial Number=5; Sample name=10_Elkin_N5_6hr_20 uM_DCVC_4-12-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034905-N SUBJECT_SAMPLE_FACTORS - S00034896 Hours of Exposure:6 hr | Treatment:No Treatment Trial Number=1; Sample name=1_Elkin_N1_6hr_NT_4-12-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034896-N SUBJECT_SAMPLE_FACTORS - S00034897 Hours of Exposure:6 hr | Treatment:No Treatment Trial Number=2; Sample name=2_Elkin_N2_6hr_NT_4-12-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034897-N SUBJECT_SAMPLE_FACTORS - S00034898 Hours of Exposure:6 hr | Treatment:No Treatment Trial Number=3; Sample name=3_Elkin_N3_6hr_NT_4-12-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034898-N SUBJECT_SAMPLE_FACTORS - S00034899 Hours of Exposure:6 hr | Treatment:No Treatment Trial Number=4; Sample name=4_Elkin_N4_6hr_NT_4-12-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034899-N SUBJECT_SAMPLE_FACTORS - S00034900 Hours of Exposure:6 hr | Treatment:No Treatment Trial Number=5; Sample name=5_Elkin_N5_6hr_NT_4-12-18; RAW_FILE_NAME=20180515-EX00848-A043-IN0001-S00034900-N #COLLECTION CO:COLLECTION_SUMMARY Standard tissue culture sample collection method CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY Exposure to cell culture media (control) or 20 µM TR:TREATMENT_SUMMARY S-(1,2-dichlorovinyl)-L-cysteine (DCVC) for 6 or 12 hours #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Standard TCS-plus assay sample preparation protocol SP:SAMPLEPREP_PROTOCOL_FILENAME SP_A043_TCA_Plus_20170713.PDF #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY HILIC NH2 chromatography CH:INSTRUMENT_NAME Agilent 1200 CH:COLUMN_NAME Phenomenex Luna NH2 (150 x 2.1mm,3um) CH:SOLVENT_A 100% water; 5mM ammonium acetate, pH 9.9, adjust with ammonium hydroxide CH:SOLVENT_B 100% acetonitrile CH:CHROMATOGRAPHY_TYPE HILIC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6520 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:MS_COMMENTS Quantitative data analysis using Agilent Quant software MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS pmol/ug MS_METABOLITE_DATA_START Samples S00034911 S00034912 S00034913 S00034914 S00034915 S00034906 S00034907 S00034908 S00034909 S00034910 S00034901 S00034902 S00034903 S00034904 S00034905 S00034896 S00034897 S00034898 S00034899 S00034900 Factors Hours of Exposure:12 hr | Treatment:20 uM DCVC Hours of Exposure:12 hr | Treatment:20 uM DCVC Hours of Exposure:12 hr | Treatment:20 uM DCVC Hours of Exposure:12 hr | Treatment:20 uM DCVC Hours of Exposure:12 hr | Treatment:20 uM DCVC Hours of Exposure:12 hr | Treatment:No Treatment Hours of Exposure:12 hr | Treatment:No Treatment Hours of Exposure:12 hr | Treatment:No Treatment Hours of Exposure:12 hr | Treatment:No Treatment Hours of Exposure:12 hr | Treatment:No Treatment Hours of Exposure:6 hr | Treatment:20 uM DCVC Hours of Exposure:6 hr | Treatment:20 uM DCVC Hours of Exposure:6 hr | Treatment:20 uM DCVC Hours of Exposure:6 hr | Treatment:20 uM DCVC Hours of Exposure:6 hr | Treatment:20 uM DCVC Hours of Exposure:6 hr | Treatment:No Treatment Hours of Exposure:6 hr | Treatment:No Treatment Hours of Exposure:6 hr | Treatment:No Treatment Hours of Exposure:6 hr | Treatment:No Treatment Hours of Exposure:6 hr | Treatment:No Treatment 2PG/3PG 27.4510 39.6060 31.7370 29.0210 38.0850 16.6790 19.4750 23.6550 29.9100 24.1940 59.7040 18.5540 27.3730 35.5190 44.2580 23.2440 20.3880 27.3620 37.9700 29.5050 6PG 4.6510 7.0150 6.7770 4.2470 5.0140 5.4790 3.5020 3.0160 3.9540 4.1160 3.9790 3.3930 4.9080 4.0180 4.3440 4.3580 4.3940 4.2140 4.3560 3.7720 aCoA 0.1420 0.3370 0.2340 0.1370 0.1920 0.1450 0.0770 0.0780 0.0800 0.0540 0.2720 0.1500 0.2600 0.1840 0.1420 0.1110 0.0820 0.1500 0.0590 0.1810 ADP 31.1650 38.6330 34.2140 26.9150 42.5760 15.0420 13.1440 12.2020 15.5120 14.9540 23.3080 17.5970 22.6290 21.6180 20.4510 12.5680 13.5450 15.4150 16.0790 19.5830 AMP 13.2440 11.4070 8.3610 5.4570 8.7120 2.5320 2.7010 2.1910 2.1860 3.3400 12.8740 8.3790 9.4700 9.1500 9.1540 2.0310 3.0570 2.6890 3.2220 3.3980 ATP 295.5940 440.1180 342.2850 271.0910 349.3340 357.4130 211.3720 241.9770 281.1910 146.9140 316.5050 237.9220 353.4370 285.7570 240.5760 302.1580 268.3730 325.2010 236.3850 321.6820 CIT/ICIT 86.7050 122.1870 125.9660 107.6790 116.9010 120.7810 92.4480 161.9040 115.2880 119.7810 100.8560 78.5490 111.8610 101.6510 97.9330 88.2860 77.0560 87.7450 96.7030 87.5100 E4P 12.6290 20.2430 19.1470 25.4870 14.8890 15.3820 11.8370 16.6950 7.7100 20.7860 24.8100 16.1670 37.8160 25.5000 25.8870 23.3110 16.5300 17.5810 16.7010 20.6010 F6P/G6P 38.5800 50.8180 43.0100 27.3080 43.2630 15.3970 13.5720 15.3330 16.2300 21.9710 22.0650 17.4470 25.1460 21.1210 22.6950 12.1570 14.4730 14.0160 16.1210 19.3210 FAD 0.0400 0.4280 0.2690 0.1110 0.1660 0.1560 0.2740 0.2030 0.2920 0.1420 0.2330 0.1980 0.3680 0.0800 0.1220 0.0750 0.2230 0.1620 0.1790 0.0170 FBP 15.3300 17.5280 12.4010 12.5570 15.5710 14.0410 11.2150 11.0340 12.5010 9.4180 18.9110 14.1720 18.5840 16.7490 15.0110 16.0400 13.4780 16.5430 15.2480 16.8190 GDP 5.7410 7.9100 6.4090 5.0470 9.5940 3.1320 2.4370 2.1910 2.7420 2.2470 4.8720 3.2050 3.8650 4.4160 3.6690 2.5130 2.5960 3.1140 2.6130 4.0020 Gl-OH3P 8.3040 10.4410 11.5880 10.4250 11.4500 8.5790 6.6250 5.9520 7.4920 5.1270 10.0240 6.2100 9.6410 7.4470 6.9540 6.4020 5.6520 5.0960 5.6290 4.1440 Glyceraldehyde3P 88.0260 124.7040 80.2500 77.1160 93.8410 48.7410 38.7230 42.8360 44.2140 49.2890 148.0530 106.6440 125.6590 117.6040 75.3420 32.5180 37.4270 46.0590 48.5420 37.1470 GMP 2.6500 2.1050 1.3390 1.1320 1.5970 0.7630 0.4950 0.6050 0.4760 0.7630 2.2990 1.5290 2.1550 1.5970 1.7690 0.5850 0.5500 0.4790 0.5470 0.7040 GTP 74.0610 104.0910 89.2280 78.4840 83.2110 87.6800 54.4770 65.2120 65.5390 48.9550 80.4850 59.6600 94.8500 74.6880 67.7220 68.5000 67.0520 74.8030 59.7390 68.2320 LAC 195.4800 201.7000 227.1700 250.0710 203.6220 204.9050 178.4740 213.6440 155.8740 246.3790 282.9180 183.2050 321.0230 230.2730 274.7250 190.2030 222.0690 192.7600 188.7580 125.7440 MAL 55.1040 69.4850 67.6350 62.1960 62.4070 47.0340 40.8320 54.5160 45.5870 50.8680 60.3260 44.2930 71.2040 54.2990 51.9520 45.1590 43.2450 46.1900 53.6120 39.2440 NAD 8.3740 12.9770 13.0520 11.6970 11.8290 12.0160 8.5370 9.8560 10.9390 9.7890 9.8830 7.3090 11.9000 8.9480 8.7670 9.2120 9.7550 9.5230 11.6960 8.2580 NADH 20.4350 23.2920 13.7170 6.4070 7.5540 9.9700 12.3080 10.0260 10.1120 7.7020 41.1780 27.6350 40.3110 33.0650 28.4580 13.7780 14.8450 14.0640 19.2540 15.4690 NADP 0.4010 0.5270 0.5280 0.5410 0.6230 0.9510 0.4930 0.5410 0.5690 0.3330 0.4310 0.3820 0.5700 0.5110 0.3340 0.7200 0.5340 0.4380 0.4300 0.4760 NADPH 1.2550 1.7240 0.5390 0.1310 0.2050 0.5170 0.1380 0.3310 0.3060 0.2590 0.9860 0.4880 0.5280 0.9560 0.4830 0.3280 0.0780 0.7120 0.5530 1.8740 PEP 1.5000 3.3830 3.1630 2.1390 2.1050 2.5340 2.1310 2.6100 3.3970 2.3820 2.4180 1.3370 3.1190 2.4160 2.0710 1.8270 2.0380 1.7480 3.3830 2.0650 R5P/X5P 5.8990 6.5740 4.2630 3.7620 3.7940 2.9610 3.6290 3.6710 2.9330 3.7450 7.8980 6.1920 9.1060 8.3820 6.9150 3.0390 3.1840 4.1660 4.3450 5.2850 s7P 2.6470 3.4630 3.0930 2.3730 2.6650 3.3100 1.7830 1.8810 2.3760 2.7740 2.3170 1.8800 2.8990 2.2860 2.9730 2.0030 2.1840 1.9690 2.5260 2.6940 SUC 79.7590 101.4660 106.1770 99.8010 85.9510 142.2460 105.4190 114.6590 120.9290 134.6100 144.9480 99.0200 151.9490 113.1550 110.1520 107.0110 129.3630 105.5710 137.7640 85.6480 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 2PG/3PG 724 6PG 91493 aCoA 444493 ADP 6022 AMP 6083 ATP 5957 CIT/ICIT 311 E4P 122357 F6P/G6P 5958 FAD 46906035 FBP 21125049 GDP 135398619 Gl-OH3P 754 Glyceraldehyde3P 729 GMP 135398631 GTP 135398633 LAC 612 MAL 525 NAD 15938971 NADH 439153 NADP 15938972 NADPH 22833512 PEP 1005 R5P/X5P 77982 s7P 165007 SUC 5988 METABOLITES_END #END