#METABOLOMICS WORKBENCH araskind_20210413_115201 DATATRACK_ID:2575 STUDY_ID:ST001743 ANALYSIS_ID:AN002836 PROJECT_ID:PR001116 VERSION 1 CREATED_ON April 15, 2021, 3:52 pm #PROJECT PR:PROJECT_TITLE The Role of Intestinal-derived FGF15 and Vertical Sleeve Gastrectomy on Plasma PR:PROJECT_TITLE Bile Acid Composition in Mice PR:PROJECT_SUMMARY Bariatric surgeries such as the Vertical Sleeve Gastrectomy (VSG) are invasive PR:PROJECT_SUMMARY but provide the most effective long-term metabolic improvements in individuals PR:PROJECT_SUMMARY with obesity and/or Type 2 diabetes. These powerful effects of manipulating the PR:PROJECT_SUMMARY gastrointestinal tract point to an important role of gastrointestinal signals in PR:PROJECT_SUMMARY regulating both energy balance and metabolism. To that end, we have used mouse PR:PROJECT_SUMMARY models of VSG to identify key gut signals that mediate these beneficial effects. PR:PROJECT_SUMMARY Previous data from our rodent model of VSG led us to hypothesize a potential PR:PROJECT_SUMMARY role for the hormone Fibroblast-Growth Factor15/19 (mouse/human ortholog) which PR:PROJECT_SUMMARY pharmacologically can regulate many aspects of energy homeostasis and glucose PR:PROJECT_SUMMARY handling. FGF15 is expressed in ileal enterocytes of the small intestine and is PR:PROJECT_SUMMARY released postprandially. Like many other gut hormones, postprandial plasma PR:PROJECT_SUMMARY concentrations of the human ortholog FGF19 and ileal FGF15 expression in mice PR:PROJECT_SUMMARY increase after VSG. We generated intestinal-specific FGF15 knock out PR:PROJECT_SUMMARY (VilCreERT2; Fgf15f/f) mice and controls, which were maintained on 60% high-fat PR:PROJECT_SUMMARY diet. VSG resulted in increased plasma bile acid levels. However, PR:PROJECT_SUMMARY intestinal-specific FGF15 knock out mice had considerably higher levels of PR:PROJECT_SUMMARY circulating total and hydrophobic bile acids after VSG. Unlike what we had PR:PROJECT_SUMMARY predicted, intestinal-specific FGF15 knock out mice lost more weight after VSG PR:PROJECT_SUMMARY as a result of increased lean tissue loss compared to control mice. Further, the PR:PROJECT_SUMMARY loss of bone mineral density and bone marrow adipose tissue observed after VSG PR:PROJECT_SUMMARY in control mice was even greater in intestinal-specific FGF15 knock out mice, PR:PROJECT_SUMMARY perhaps secondary to anemia and elevated erythropoietin/FGF23. Finally the PR:PROJECT_SUMMARY effect of VSG to improve glucose tolerance and to reduce hepatic cholesterol was PR:PROJECT_SUMMARY also absent in intestinal-specific FGF15 knock out mice. These data point to an PR:PROJECT_SUMMARY important role for intestinal FGF15 to protect the organism from deleterious PR:PROJECT_SUMMARY effects of bile acid toxicity after VSG. PR:INSTITUTE University of Michigan PR:DEPARTMENT MRC2 PR:LABORATORY Metabolomics core PR:LAST_NAME Kachman PR:FIRST_NAME Maureen PR:ADDRESS 1000 Wall St., 5458 PR:EMAIL mkachman@med.umich.edu PR:PHONE 734-232-0842 PR:FUNDING_SOURCE NIH: 1U2CDK110678-01, P30-AR069620, P30 DK089503, DK097153, 5T32DK108740, PR:FUNDING_SOURCE 5T32DK071212-12, UL1TR002240, DK020572, DK089503, DK107282, DK121995, RO1 PR:FUNDING_SOURCE DK62876 , R24 DK092759, American Diabetes Association grants 1-19-IBS-252 and PR:FUNDING_SOURCE 1-18-PDF-087 #STUDY ST:STUDY_TITLE The Role of Intestinal-derived FGF15 and Vertical Sleeve Gastrectomy on Plasma ST:STUDY_TITLE Bile Acid Composition in Mice ST:STUDY_TYPE MS analysis ST:STUDY_SUMMARY Bariatric surgeries such as the Vertical Sleeve Gastrectomy (VSG) are invasive ST:STUDY_SUMMARY but provide the most effective long-term metabolic improvements in individuals ST:STUDY_SUMMARY with obesity and/or Type 2 diabetes. These powerful effects of manipulating the ST:STUDY_SUMMARY gastrointestinal tract point to an important role of gastrointestinal signals in ST:STUDY_SUMMARY regulating both energy balance and metabolism. To that end, we have used mouse ST:STUDY_SUMMARY models of VSG to identify key gut signals that mediate these beneficial effects. ST:STUDY_SUMMARY Previous data from our rodent model of VSG led us to hypothesize a potential ST:STUDY_SUMMARY role for the hormone Fibroblast-Growth Factor15/19 (mouse/human ortholog) which ST:STUDY_SUMMARY pharmacologically can regulate many aspects of energy homeostasis and glucose ST:STUDY_SUMMARY handling. FGF15 is expressed in ileal enterocytes of the small intestine and is ST:STUDY_SUMMARY released postprandially. Like many other gut hormones, postprandial plasma ST:STUDY_SUMMARY concentrations of the human ortholog FGF19 and ileal FGF15 expression in mice ST:STUDY_SUMMARY increase after VSG. We generated intestinal-specific FGF15 knock out ST:STUDY_SUMMARY (VilCreERT2; Fgf15f/f) mice and controls, which were maintained on 60% high-fat ST:STUDY_SUMMARY diet. VSG resulted in increased plasma bile acid levels. However, ST:STUDY_SUMMARY intestinal-specific FGF15 knock out mice had considerably higher levels of ST:STUDY_SUMMARY circulating total and hydrophobic bile acids after VSG. Unlike what we had ST:STUDY_SUMMARY predicted, intestinal-specific FGF15 knock out mice lost more weight after VSG ST:STUDY_SUMMARY as a result of increased lean tissue loss compared to control mice. Further, the ST:STUDY_SUMMARY loss of bone mineral density and bone marrow adipose tissue observed after VSG ST:STUDY_SUMMARY in control mice was even greater in intestinal-specific FGF15 knock out mice, ST:STUDY_SUMMARY perhaps secondary to anemia and elevated erythropoietin/FGF23. Finally the ST:STUDY_SUMMARY effect of VSG to improve glucose tolerance and to reduce hepatic cholesterol was ST:STUDY_SUMMARY also absent in intestinal-specific FGF15 knock out mice. These data point to an ST:STUDY_SUMMARY important role for intestinal FGF15 to protect the organism from deleterious ST:STUDY_SUMMARY effects of bile acid toxicity after VSG. ST:INSTITUTE University of Michigan ST:DEPARTMENT Surgery ST:LABORATORY Seeley Lab ST:LAST_NAME Seeley ST:FIRST_NAME Randy ST:ADDRESS Ann Arbor, MI, 48105, USA ST:EMAIL seeleyrj@umich.edu ST:PHONE 734-615-2880 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENDER Male #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 380 S00046232 Genotype:FGF15INT-KO | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046232-1-N.d SUBJECT_SAMPLE_FACTORS 407 S00046233 Genotype:CTRL | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046233-2-N.d SUBJECT_SAMPLE_FACTORS 342 S00046234 Genotype:CTRL | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046234-3-N.d SUBJECT_SAMPLE_FACTORS 463 S00046235 Genotype:FGF15INT-KO | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046235-4-N.d SUBJECT_SAMPLE_FACTORS 464 S00046236 Genotype:FGF15INT-KO | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046236-5-N.d SUBJECT_SAMPLE_FACTORS 496 S00046237 Genotype:CTRL | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046237-6-N.d SUBJECT_SAMPLE_FACTORS 469 S00046238 Genotype:FGF15INT-KO | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046238-7-N.d SUBJECT_SAMPLE_FACTORS 498 S00046239 Genotype:CTRL | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046239-8-N.d SUBJECT_SAMPLE_FACTORS 479 S00046240 Genotype:CTRL | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046240-9-N.d SUBJECT_SAMPLE_FACTORS 340 S00046241 Genotype:CTRL | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046241-10-N.d SUBJECT_SAMPLE_FACTORS 471 S00046242 Genotype:CTRL | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046242-11-N.d SUBJECT_SAMPLE_FACTORS 501 S00046244 Genotype:FGF15INT-KO | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046244-13-N.d SUBJECT_SAMPLE_FACTORS 412 S00046245 Genotype:CTRL | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046245-14-N.d SUBJECT_SAMPLE_FACTORS 472 S00046247 Genotype:FGF15INT-KO | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046247-16-N.d SUBJECT_SAMPLE_FACTORS 429 S00046248 Genotype:CTRL | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046248-17-N.d SUBJECT_SAMPLE_FACTORS 435 S00046249 Genotype:FGF15INT-KO | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046249-18-N.d SUBJECT_SAMPLE_FACTORS 478 S00046250 Genotype:CTRL | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046250-19-N.d SUBJECT_SAMPLE_FACTORS 474 S00046251 Genotype:FGF15INT-KO | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046251-20-N.d SUBJECT_SAMPLE_FACTORS 468 S00046252 Genotype:CTRL | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046252-21-N.d SUBJECT_SAMPLE_FACTORS 465 S00046253 Genotype:FGF15INT-KO | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046253-22-N.d SUBJECT_SAMPLE_FACTORS 485 S00046254 Genotype:CTRL | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046254-23-N.d SUBJECT_SAMPLE_FACTORS 499 S00046256 Genotype:FGF15INT-KO | Surgery:VSG RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046256-25-N.d SUBJECT_SAMPLE_FACTORS 500 S00046257 Genotype:FGF15INT-KO | Surgery:Sham RAW_FILE_NAME=20200827-EX01051-A014-IN0026-S00046257-26-N.d #COLLECTION CO:COLLECTION_SUMMARY Mouse blood collection CO:SAMPLE_TYPE Blood (plasma) #TREATMENT TR:TREATMENT_SUMMARY HFD diet after sham or VSG surgery #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Standard bile acid analysis sample prep SP:SAMPLEPREP_PROTOCOL_FILENAME Bile_Acids_Assay.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 6410 CH:COLUMN_NAME Waters XBridge C18 (150mm x 2.1mm, 3.5um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6410 QQQ MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Agilent Data Acquisition/Agilent Quant #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nM MS_METABOLITE_DATA_START Samples S00046232 S00046233 S00046234 S00046235 S00046236 S00046237 S00046238 S00046239 S00046240 S00046241 S00046242 S00046244 S00046245 S00046247 S00046248 S00046249 S00046250 S00046251 S00046252 S00046253 S00046254 S00046256 S00046257 Factors Genotype:FGF15INT-KO | Surgery:VSG Genotype:CTRL | Surgery:Sham Genotype:CTRL | Surgery:VSG Genotype:FGF15INT-KO | Surgery:VSG Genotype:FGF15INT-KO | Surgery:Sham Genotype:CTRL | Surgery:VSG Genotype:FGF15INT-KO | Surgery:Sham Genotype:CTRL | Surgery:VSG Genotype:CTRL | Surgery:Sham Genotype:CTRL | Surgery:VSG Genotype:CTRL | Surgery:VSG Genotype:FGF15INT-KO | Surgery:Sham Genotype:CTRL | Surgery:Sham Genotype:FGF15INT-KO | Surgery:Sham Genotype:CTRL | Surgery:Sham Genotype:FGF15INT-KO | Surgery:VSG Genotype:CTRL | Surgery:VSG Genotype:FGF15INT-KO | Surgery:Sham Genotype:CTRL | Surgery:Sham Genotype:FGF15INT-KO | Surgery:VSG Genotype:CTRL | Surgery:VSG Genotype:FGF15INT-KO | Surgery:VSG Genotype:FGF15INT-KO | Surgery:Sham tauro-alpha-muricholate 1419.700544 377.2009929 286.7276803 3605.566435 786.8351004 1351.526307 800.94723 652.9623288 216.001135 722.5893938 2266.500697 146.9720861 572.6002415 226.5612639 293.8860259 1101.995654 33.36943616 94.63014674 114.3354768 1811.065079 747.5390812 1602.29818 750.0739437 Tauro-gamma-muricholate 0.124147139 1.490634564 0.190672729 14.50812133 2.252225063 2.537164374 0.555846773 0.035376208 0.519097651 0.22188 3.628924593 0.316412519 2.566983377 0.313555241 3.368084725 6.437736482 0.780415214 2.521067031 1.469671587 3.174676214 0.718543053 1.702883703 1.816858779 tauroursodeoxycholate 11.98828893 9.606781724 7.568487916 16.51747905 14.54553437 23.14152388 7.918186181 16.85207312 6.134277243 15.26038897 29.12253252 3.795379461 8.295628534 6.015476248 8.950719963 17.69607173 4.273649804 8.259375934 5.373759749 11.6024981 18.74967886 21.31554073 20.18098229 taurohyodeoxycholate 1.100902228 5.414241621 3.897862355 55.20290969 5.720033363 3.205175211 4.740378591 3.598294066 5.895165401 5.746231713 2.426784604 3.011297987 1.557812099 1.718581407 1.854379155 14.45602626 3.776806344 2.887388817 3.584586075 3.623966889 3.538465224 2.036907125 21.78375443 Taurocholate 177.4837267 70.81890958 16.22190326 2226.190468 957.4146501 35.71537064 289.217774 33.88485295 57.59546176 77.92021428 82.64844117 204.1376835 23.91179521 217.9179339 61.5385473 903.3702478 11.30907071 101.0426056 62.79207595 712.8638463 68.60785961 391.5734246 501.8765685 omega-Muricholate 9.273287777 15.99987693 9.187671902 70.45199697 0.593025305 13.34230866 2.077619232 12.86072907 2.835307145 14.35633954 18.2192282 2.864938543 26.23986306 1.70129629 11.80265097 33.35202287 2.871547957 2.764660441 11.01785555 27.2912475 61.27164449 19.71296733 17.0158253 glycoursodeoxycholate 0.027027426 0.04689151 0.048368189 0.041155287 0.077267622 0.05404364 0.051022886 0.057403847 0.030828416 0.061520414 0.043644334 0.054598438 0.059455481 0.016789026 0.088146098 0.054691958 0.010884796 0.017665529 0.041629358 0.037659168 0.038918312 0.125201175 0.054819534 Glycocholate 8.708272147 4.317007468 0.450650391 33.55190036 5.201024445 1.325340891 9.036195264 2.681654023 1.335732961 2.163321797 2.938376629 4.185203903 3.78127765 1.765813413 2.956661365 13.26873436 0.571728004 3.455781192 0.456121581 11.16191368 1.26018093 16.86022754 15.56594998 glycohyodeoxycholate 0.012207936 0.006190037 0.034370898 0.012692041 0.01147552 0.005994402 0.010984159 0.009488564 0.006921015 0.009001181 0.007112341 0.042620361 0.037449032 0.006260269 0.041415611 0.006990411 0.008457995 0.017094997 0.012114398 0.014096979 0.018789509 0.011089671 0.036249996 alpha-Murocholate 7.902205431 8.639549688 4.015221345 6.837864111 5.593305235 11.00259174 10.19614133 6.977086259 4.744871767 4.918790873 5.448683117 3.825981889 6.861377669 4.421740131 8.501781754 11.46699174 2.988252285 8.540495832 13.44493929 6.119872491 5.506333258 15.12067399 9.258706832 beta-Muricholate 137.9674976 14.19244186 27.31668179 21.6083151 1.778866448 34.80710066 6.752583486 96.52860253 5.253372421 55.60643011 54.73940207 15.23876424 66.81193071 4.535064372 10.13464432 31.02352176 9.814173406 14.97209782 21.86192642 67.50386582 103.1927696 77.22869148 13.33523145 taurochenodeoxycholate 2.796538211 4.451912987 1.888899049 10.77969663 18.56202039 0.843615797 5.908363105 2.560038687 1.185198857 2.004665644 8.591312637 4.66476975 0.665579523 6.080358676 2.188506542 7.568275124 1.144906131 2.266665475 1.022478145 2.657847891 4.580221003 2.122051423 10.96978429 taurodeoxycholate 47.79425501 34.34514897 9.555776881 527.5259615 391.9605608 3.911490012 236.2110906 37.41562115 5.749029752 71.71117096 25.90634903 110.6371134 3.479502793 123.991556 10.55027195 468.3986793 13.11156781 106.2521783 16.02817042 109.8634567 46.50834222 40.56434995 608.5028942 hyocholicacid 4.015223932 4.14311287 3.404386405 6.562333981 4.544536766 2.239339537 2.056018233 2.93451351 2.325371467 2.402475003 3.381434089 1.705968795 4.150111663 3.475456854 1.895153682 1.930983673 2.163685195 2.207368649 2.496586861 1.364541557 3.008525319 3.922522383 2.569999028 Cholate 727.4780004 188.2879065 6.635251662 274.4126551 6.961891068 37.30503197 94.90338043 45.44419522 16.94423456 80.97517657 31.92556406 1.182956956 90.46518306 20.03537937 69.16311418 225.2406616 11.36869438 19.06128679 12.18258925 76.47441279 56.73689858 489.0916167 52.55964202 ursodeoxycholate 5.484319242 5.141945443 2.440568814 1.738879295 0.084768345 7.84360206 1.154364035 2.481167295 0.606316357 6.519953679 4.904443706 0.415012289 12.13692268 5.595127686 5.305235672 3.365775113 1.494420812 0.589555034 2.181658889 4.666348145 13.99036239 9.063998551 5.900605497 glycochenodeoxycholate 0.210259839 0.251370839 0.065437136 0.061631345 0.088695221 0.025135469 0.047544611 0.094880877 0.027356638 0.017942147 0.021832499 0.126183212 0.084497752 0.033553243 0.071213762 0.17588117 0.038276661 0.023303474 0.048719383 0.052798283 0.031143699 0.078481799 0.226196027 hyodeoxycholate 1.153262025 0.1565801 0.968762551 11.65143655 0.768422571 0.029549433 0 0.881210289 0 3.332947155 0 1.445491873 0 1.586721883 0.684892925 5.155462205 0.601364432 0.223362294 0 0.816262753 3.977630874 0.652817971 3.953285896 glycodeoxycholate 0.090991725 0.007854669 0.012470377 0.336552411 0.040876904 0.027937648 0.157205555 0.031944446 0.04000144 0.067937841 0.00933584 0.069514955 0.02219091 0.086411167 0.052446834 0.154586598 0.011612867 0.051767186 0.031799766 0.026071516 0.028143331 0.045170157 0.387111608 Taurolithocholate 0.217040123 2.051709006 0.949459267 1.116335105 1.186514359 0.244744491 1.368662132 0.781695908 0.188724766 0.377282635 1.158773144 1.135051783 0.249399285 0.668240233 0.325252344 0.519207749 0.269809546 0.88786146 0.193166547 0.463927631 0.527035496 0.206659394 2.445557946 chenodeoxycholate 215.7432966 41.45734403 19.34419108 516.5620619 100.9054642 46.45923275 110.5032366 139.5591587 10.72507969 203.4612399 97.34951959 73.95294134 255.5853442 198.4480882 28.0293453 424.6010649 6.684164496 118.4880783 34.06135018 468.5615322 316.8727029 261.2022549 474.5733304 Deoxycholate 223.6156944 44.2491309 20.05005388 535.4112318 104.5874695 48.15451398 114.5354613 144.6516238 9.814179129 210.8854698 100.9017697 76.65145844 57.96853375 205.6893899 29.05212636 440.09461 6.835941677 122.8116671 35.30423699 485.6591794 328.4352776 270.7334343 491.8903461 glycolithocholate 5.950503357 8.095289974 4.905035554 23.80437619 9.398117078 2.49512507 0.862280136 2.739819568 22.42217185 9.781768671 16.6981688 18.33553116 3.795913245 1.180092442 4.592093845 20.53103553 28.57527751 22.97498751 2.315033981 17.10617906 12.75493715 25.16332441 21.18569702 lithocholate 3.958902324 3.920048019 12.5778321 21.70201096 9.270124401 43.78715225 24.03888445 18.64341791 27.89944561 2.430945208 18.57554289 7.390393123 4.07192359 8.43310047 18.82761121 17.83780317 8.436437114 3.309974737 2.95277125 22.25177781 5.205187963 3.505569621 18.96787516 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name PubChem tauro-alpha-muricholate 101657566 Tauro-gamma-muricholate 11954195 tauroursodeoxycholate 9848818 taurohyodeoxycholate 119046 Taurocholate 6675 omega-Muricholate 5283851 glycoursodeoxycholate 12310288 Glycocholate 10140 glycohyodeoxycholate 114611 alpha-Murocholate 5283852 beta-Muricholate 5283853 taurochenodeoxycholate 387316 taurodeoxycholate 2733768 hyocholicacid 92805 Cholate 221493 ursodeoxycholate 31401 glycochenodeoxycholate 12544 hyodeoxycholate 5283820 glycodeoxycholate 3035026 Taurolithocholate 439763 chenodeoxycholate 10133 Deoxycholate 222528 glycolithocholate 115245 lithocholate 9903 METABOLITES_END #END