#METABOLOMICS WORKBENCH araskind_20200609_095708 DATATRACK_ID:2050 STUDY_ID:ST001398 ANALYSIS_ID:AN002338 PROJECT_ID:PR000958 VERSION 1 CREATED_ON June 9, 2020, 2:55 pm #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 #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 #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]Raw file names and additional sample data 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 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 - 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 - 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 #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_ID SP_A043_TCA_Plus_20170713.PDF #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY HILIC NH2 chromatography CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent 1200 CH:COLUMN_NAME Phenomenex Luna NH2 (150 x 2.1mm, 3um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6520 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Semi-quantitative data analysis using Agilent Quant software #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS area/ug protein MS_METABOLITE_DATA_START Samples S00034896 S00034897 S00034898 S00034899 S00034900 S00034901 S00034902 S00034903 S00034904 S00034905 S00034906 S00034907 S00034908 S00034909 S00034910 S00034911 S00034912 S00034913 S00034914 S00034915 Factors 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 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: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: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 3-Phospho-serine 13.292 15.945 13.970 17.792 12.185 16.666 11.781 18.374 14.343 12.559 15.673 11.002 12.266 14.337 11.865 14.631 20.398 17.523 15.033 17.467 Acetylphosphate 1.272 2.080 1.468 1.902 1.074 2.504 1.312 2.248 1.498 1.621 1.703 1.381 1.461 1.544 1.773 1.440 2.133 1.883 1.544 1.675 Alanine 15.139 19.014 16.044 17.854 14.195 28.049 20.663 33.601 25.486 27.571 20.153 15.020 16.864 16.803 21.701 25.941 28.411 30.979 27.317 25.565 Arginine 59.234 85.009 68.474 65.242 41.981 94.464 63.217 116.209 77.733 90.589 75.452 61.877 80.471 54.227 87.737 68.442 74.553 81.300 97.086 73.435 Asparagine 46.517 59.488 50.392 64.150 34.411 69.129 46.019 85.170 55.511 64.766 47.650 48.501 63.457 55.536 76.745 51.015 61.280 70.894 75.808 58.518 Aspartate 20.546 26.985 22.245 24.143 16.969 25.328 19.763 31.840 23.045 24.626 26.318 20.393 23.222 18.606 23.068 22.509 26.063 26.613 26.334 23.214 Citrulline 4.444 7.082 5.249 5.897 3.025 8.508 5.985 10.739 7.223 8.089 5.616 5.620 7.529 5.126 9.135 5.555 5.876 7.634 10.951 5.803 CMP 0.412 0.625 0.457 0.411 0.430 0.810 0.646 0.885 0.712 0.714 0.463 0.404 0.472 0.490 0.511 0.850 0.997 0.782 0.559 0.667 Creatine 8.977 12.535 7.757 13.684 8.192 18.667 13.855 21.050 16.247 16.152 10.771 9.812 9.703 11.290 15.202 16.571 19.265 21.882 18.577 14.309 Creatinine 3.367 4.345 3.641 3.651 2.588 4.803 3.617 6.446 4.701 4.988 4.596 3.422 4.314 3.236 4.804 4.267 4.464 4.749 5.368 4.234 Deoxyuridine 2.032 2.012 2.393 1.772 2.266 2.307 1.830 3.371 2.351 2.935 2.494 1.289 1.678 1.260 1.360 2.047 2.038 1.875 2.343 2.457 DHAP 0.235 0.276 0.252 0.286 0.189 0.736 0.567 0.665 0.653 0.419 0.313 0.297 0.312 0.225 0.284 0.508 0.658 0.526 0.455 0.460 Gluconate 28.172 30.483 26.534 34.230 23.154 35.973 29.191 43.327 32.720 32.862 32.416 24.432 29.663 29.576 29.188 32.165 41.316 39.937 36.432 35.404 Glutamate 137.479 150.199 135.054 175.008 117.902 171.107 137.513 212.772 161.297 160.159 165.870 135.789 148.072 155.480 151.253 170.582 217.704 215.479 186.551 182.677 Glutamine 569.909 879.243 680.128 764.814 456.449 1035.401 702.146 1240.761 880.659 996.157 653.549 594.470 747.647 588.144 931.294 724.545 794.966 913.020 1038.594 738.229 Glutathione-oxidized 13.993 13.908 12.465 13.623 10.578 15.827 12.151 20.685 14.725 15.902 14.944 9.471 10.806 10.370 11.382 13.294 16.384 16.569 17.456 14.320 Glutathione-Reduced 52.439 59.734 63.848 87.102 72.327 87.850 68.322 88.292 81.276 60.404 89.333 73.478 79.346 102.720 76.002 112.891 161.343 142.363 89.018 118.909 Hexose (glucose etc.) 0.711 0.691 0.442 0.664 0.546 0.808 0.488 0.743 0.314 0.698 0.188 0.181 0.327 0.279 0.228 0.194 0.339 0.220 0.242 0.372 Histidine 1.224 1.174 1.011 1.069 0.802 1.892 1.175 1.770 1.368 1.249 1.649 1.322 1.149 1.480 0.921 1.626 2.057 2.228 2.048 2.171 Homocysteic acid 0.363 0.393 0.325 0.327 0.255 0.243 0.224 0.216 0.167 0.183 0.488 0.241 0.402 0.232 0.258 0.290 0.265 0.245 0.343 0.300 Hypoxanthine 28.130 36.369 29.823 34.750 23.145 40.327 28.287 43.993 33.865 31.974 33.737 28.336 30.844 30.834 33.524 31.257 36.475 38.131 36.692 31.207 IMP 0.423 0.586 0.482 0.533 0.454 0.876 0.785 1.351 0.925 0.758 0.296 0.239 0.281 0.288 0.350 0.540 0.529 0.433 0.522 0.252 Inosine 0.306 0.709 0.526 0.525 0.381 18.840 6.114 8.950 6.694 7.633 0.348 0.399 0.716 0.693 1.122 2.670 2.013 1.108 0.902 3.173 Leucine/isoleucine 369.467 462.662 389.992 402.548 279.373 549.120 400.282 689.775 489.859 538.455 431.999 355.541 429.893 334.852 488.616 448.282 498.960 541.513 603.620 483.225 Lysine 8.222 13.106 9.544 10.453 5.292 14.527 10.059 18.874 12.651 15.830 10.644 9.098 12.525 7.691 14.965 10.442 10.134 12.350 16.902 10.522 Methionine 14.107 18.723 15.645 15.753 10.633 21.518 14.612 25.843 18.188 20.402 15.758 12.889 16.064 12.893 18.283 16.104 17.591 18.979 21.598 17.586 N-Ac-glucosamine-1-P 0.232 0.401 0.414 0.546 0.682 1.249 0.827 1.215 1.073 1.539 0.264 0.343 1.426 0.421 0.555 0.994 1.237 0.841 0.683 0.671 Oleic acid 18.919 18.257 20.319 33.577 37.636 167.240 81.364 103.874 88.806 84.883 25.482 18.149 23.007 24.148 44.995 157.257 138.642 90.490 59.175 94.840 Ornithine 18.428 25.899 21.307 20.327 12.710 29.579 19.841 36.748 25.256 29.369 24.872 20.245 26.978 17.276 29.383 23.165 24.860 27.784 33.412 23.623 Palmitic acid 228.033 172.995 175.131 189.217 137.260 295.165 152.346 214.204 174.400 167.558 175.404 122.871 168.247 113.564 125.307 185.186 188.867 160.286 190.966 151.688 Pantothenate 140.238 165.479 142.467 165.582 93.110 215.408 144.411 252.360 172.107 190.294 146.051 146.479 166.283 145.605 197.851 139.370 171.246 181.713 200.937 147.567 Phenylalanine 60.176 78.625 66.112 68.374 45.471 91.435 62.575 108.782 76.039 84.717 67.444 55.386 68.724 53.901 78.117 67.678 78.157 81.655 93.175 75.525 Phenylpyruvate 0.945 1.050 0.944 0.912 0.716 1.280 0.900 1.441 1.108 1.254 1.123 0.827 1.068 0.834 1.189 1.073 1.181 1.267 1.285 1.199 Phosphocreatine 4.811 4.855 4.266 5.543 3.995 3.813 2.984 4.858 3.723 3.139 4.920 3.828 4.602 4.878 4.167 1.788 2.911 2.935 2.589 2.580 Proline 32.995 32.784 30.780 31.076 29.256 32.932 26.220 38.446 32.235 29.137 40.106 30.350 31.906 31.671 29.815 31.766 36.410 37.367 33.126 33.405 Serine 0.257 0.190 0.324 0.475 0.385 0.725 0.217 0.295 0.440 0.535 0.157 0.228 0.287 0.362 0.291 0.336 0.451 0.366 0.339 0.449 Stearic acid 0.894 0.915 0.887 1.022 0.920 0.785 0.801 1.052 0.954 0.919 1.285 0.685 0.665 0.964 1.034 1.111 1.610 1.600 0.932 1.087 Taurine 0.250 0.231 0.227 0.254 0.249 0.469 0.308 0.477 0.410 0.329 0.289 0.264 0.205 0.241 0.228 0.342 0.467 0.479 0.328 0.384 Threonine 7.939 8.946 8.891 8.164 7.509 10.264 7.790 12.615 8.734 8.933 10.748 8.065 9.331 9.296 8.143 9.026 12.372 11.887 11.453 9.298 Tryptophan 0.892 1.475 1.218 1.527 1.628 6.895 4.524 4.958 4.832 4.851 1.143 1.361 0.980 0.944 1.696 7.399 6.169 4.327 2.697 4.563 Tyrosine 42.896 40.884 46.237 33.661 45.235 44.995 33.708 49.426 39.405 33.774 49.886 31.463 35.129 40.066 21.849 43.001 62.353 49.321 40.247 49.400 UDP 11.990 11.491 12.625 12.337 11.656 14.589 12.074 20.358 14.560 14.210 17.146 11.837 22.425 15.376 13.930 12.104 18.017 16.645 13.687 15.184 UDP-D-glucose 1.121 0.944 1.024 1.033 1.060 1.208 0.987 1.573 1.059 1.012 1.149 0.872 1.013 1.164 0.969 1.098 1.595 1.480 1.156 1.326 UDP-D-glucuronate 0.138 0.035 0.052 0.029 0.099 0.064 0.128 0.173 0.138 0.115 0.115 0.029 0.091 0.015 0.061 0.081 0.037 0.019 0.028 0.012 UDP-N-Ac-D-glucosamine 2.676 3.539 3.189 4.100 4.694 4.998 4.554 6.466 5.448 5.634 3.851 3.244 4.126 4.063 5.689 9.568 12.050 10.269 7.012 10.067 UMP 0.039 0.108 0.106 0.092 0.008 0.121 0.105 0.187 0.041 0.063 0.095 0.141 0.109 0.156 0.086 0.032 0.228 0.131 0.058 0.096 UTP 1.273 1.029 1.381 1.277 1.408 1.502 1.268 1.616 1.603 1.219 1.238 0.901 1.069 1.058 0.804 1.269 1.400 0.914 1.028 1.201 Valine 0.823 1.444 1.627 1.413 0.600 1.847 1.101 2.686 0.840 1.880 1.947 0.903 1.682 1.079 1.565 1.007 2.532 1.301 2.165 2.076 Xanthine 0.415 0.342 0.268 0.254 0.452 0.973 0.024 0.776 0.027 0.513 0.175 0.279 0.484 0.178 0.341 0.612 0.842 0.415 0.448 0.395 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name PubChem CID 3-Phospho-serine 106 Acetylphosphate 186 Alanine 602 Arginine 6322 Asparagine 6267 Aspartate 5960 Citrulline 9750 CMP 6131 Creatine 586 Creatinine 588 Deoxyuridine 13712 DHAP 668 Gluconate 10690 Glutamate 33032 Glutamine 5961 Glutathione-oxidized 975 Glutathione-Reduced 124886 Hexose (glucose etc.) 206 Histidine 6274 Homocysteic acid 92117 Hypoxanthine 135398638 IMP 135398640 Inosine 135398641 Leucine/isoleucine 6106 Lysine 5962 Methionine 876 N-Ac-glucosamine-1-P 440272 Oleic acid 445639 Ornithine 6262 Palmitic acid 985 Pantothenate 6613 Phenylalanine 6140 Phenylpyruvate 997 Phosphocreatine 9548602 Proline 145742 Serine 5951 Stearic acid 5281 Taurine 1123 Threonine 6288 Tryptophan 6305 Tyrosine 6057 UDP 6031 UDP-D-glucose 439156 UDP-D-glucuronate 17473 UDP-N-Ac-D-glucosamine 445675 UMP 6030 UTP 6133 Valine 6287 Xanthine 1188 METABOLITES_END #END