#METABOLOMICS WORKBENCH araskind_20191211_122141 DATATRACK_ID:1866 STUDY_ID:ST001287 ANALYSIS_ID:AN002148 PROJECT_ID:PR000869 VERSION 1 CREATED_ON December 17, 2019, 12:59 pm #PROJECT PR:PROJECT_TITLE Luminal succinate in UC-HMA (human microbiota-associated) mice. PR:PROJECT_TYPE MS analysis PR:PROJECT_SUMMARY Luminal succinate level in UC-HMA (UC patient derived microbiota colonized HMA) PR:PROJECT_SUMMARY mice, C. difficile infection susceptible mice. Patients with ulcerative colitis PR:PROJECT_SUMMARY (UC) are known to be at higher risk for Clostridium difficile (C. difficile) PR:PROJECT_SUMMARY infection (CDI), and CDI in UC patients is recognized as a major clinical PR:PROJECT_SUMMARY problem because it worsens UC outcome. In order to assess the role of gut PR:PROJECT_SUMMARY dysbiosis, seen in UC patients, we have established a human PR:PROJECT_SUMMARY microbiota-associated mouse model in which germ-free mice are colonized with gut PR:PROJECT_SUMMARY microbiota from UC patients. Utilizing this model, we found that UC microbiota PR:PROJECT_SUMMARY colonized HMA mice (UC-HMA mice) were susceptible to CDI. To address the PR:PROJECT_SUMMARY mechanism by which UC-HMA mice are unable to acquire the colonization resistance PR:PROJECT_SUMMARY against C. difficile, we analyzed the luminal metabolites in UC-HMA mice, PR:PROJECT_SUMMARY especially in succinate which is a crucial metabolite doe the growth of C. PR:PROJECT_SUMMARY difficile. PR:INSTITUTE University of Michigan PR:DEPARTMENT Internal Medicine-Gastroenterology PR:LABORATORY Kamada Lab PR:LAST_NAME Kamada PR:FIRST_NAME Nobuhiko PR:ADDRESS Ann Arbor, MI PR:EMAIL nkamada@umich.edu PR:PHONE 734-763-2142 #STUDY ST:STUDY_TITLE Luminal succinate in UC-HMA (human microbiota-associated) mice. ST:STUDY_TYPE MS analysis ST:STUDY_SUMMARY Patients with ulcerative colitis (UC) are known to be at higher risk for ST:STUDY_SUMMARY Clostridium difficile (C. difficile) infection (CDI), and CDI in UC patients is ST:STUDY_SUMMARY recognized as a major clinical problem because it worsens UC outcome. In order ST:STUDY_SUMMARY to assess the role of gut dysbiosis, seen in UC patients, we have established a ST:STUDY_SUMMARY human microbiota-associated mouse model in which germ-free mice are colonized ST:STUDY_SUMMARY with gut microbiota from UC patients. Utilizing this model, we found that UC ST:STUDY_SUMMARY microbiota colonized HMA mice (UC-HMA mice) were susceptible to CDI. To address ST:STUDY_SUMMARY the mechanism by which UC-HMA mice are unable to acquire the colonization ST:STUDY_SUMMARY resistance against C. difficile, we analyzed the luminal metabolites in UC-HMA ST:STUDY_SUMMARY mice, especially in succinate which is a crucial metabolite doe the growth of C. ST:STUDY_SUMMARY difficile. 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:NUM_GROUPS 63 ST:TOTAL_SUBJECTS 3 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN UC-hGB #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 SU0024964 S00034622 Replication:#1 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_027.d SUBJECT_SAMPLE_FACTORS SU0024965 S00034623 Replication:#1 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_036.d SUBJECT_SAMPLE_FACTORS SU0024966 S00034624 Replication:#1 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_025.d SUBJECT_SAMPLE_FACTORS SU0024967 S00034625 Replication:#1 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_017.d SUBJECT_SAMPLE_FACTORS SU0024968 S00034626 Replication:#1 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_018.d SUBJECT_SAMPLE_FACTORS SU0024969 S00034627 Replication:#2 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_038.d SUBJECT_SAMPLE_FACTORS SU0024970 S00034628 Replication:#2 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_016.d SUBJECT_SAMPLE_FACTORS SU0024971 S00034629 Replication:#2 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_021.d SUBJECT_SAMPLE_FACTORS SU0024972 S00034630 Replication:#2 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_023.d SUBJECT_SAMPLE_FACTORS SU0024973 S00034631 Replication:#2 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_019.d SUBJECT_SAMPLE_FACTORS SU0024974 S00034632 Replication:#2 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_033.d SUBJECT_SAMPLE_FACTORS SU0024975 S00034633 Replication:#2 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_012.d SUBJECT_SAMPLE_FACTORS SU0024976 S00034634 Replication:#2 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_028.d SUBJECT_SAMPLE_FACTORS SU0024977 S00034635 Replication:#2 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_041.d SUBJECT_SAMPLE_FACTORS SU0024978 S00034636 Replication:#2 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_010.d SUBJECT_SAMPLE_FACTORS SU0024979 S00034637 Replication:#3 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_029.d SUBJECT_SAMPLE_FACTORS SU0024980 S00034638 Replication:#3 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_034.d SUBJECT_SAMPLE_FACTORS SU0024981 S00034639 Replication:#3 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_035.d SUBJECT_SAMPLE_FACTORS SU0024982 S00034640 Replication:#3 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_030.d SUBJECT_SAMPLE_FACTORS SU0024983 S00034641 Replication:#3 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_039.d SUBJECT_SAMPLE_FACTORS SU0024984 S00034642 Replication:#3 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_014.d SUBJECT_SAMPLE_FACTORS SU0024985 S00034643 Replication:#3 | Treatment:Ctrl RAW_FILE_NAME=2018-04-20_026.d SUBJECT_SAMPLE_FACTORS SU0024986 S00034644 Replication:#3 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_037.d SUBJECT_SAMPLE_FACTORS SU0024987 S00034645 Replication:#3 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_013.d SUBJECT_SAMPLE_FACTORS SU0024988 S00034646 Replication:#3 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_040.d SUBJECT_SAMPLE_FACTORS SU0024989 S00034647 Replication:#3 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_024.d SUBJECT_SAMPLE_FACTORS SU0024990 S00034648 Replication:#3 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_015.d SUBJECT_SAMPLE_FACTORS SU0024991 S00034649 Replication:#3 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_011.d SUBJECT_SAMPLE_FACTORS SU0024992 S00034650 Replication:#3 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_032.d SUBJECT_SAMPLE_FACTORS SU0024993 S00034651 Replication:#3 | Treatment:SuccinateConsumingBacteria RAW_FILE_NAME=2018-04-20_022.d #COLLECTION CO:COLLECTION_SUMMARY Mouse feces CO:SAMPLE_TYPE Feces #TREATMENT TR:TREATMENT_SUMMARY None #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 2018-4-20_Succinate_fecal_EX00837.docx #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent Q-TOF 6520 CH:COLUMN_NAME Phenomenex Luna NH2, 1mm ID CH:METHODS_FILENAME 002-HILIC-27min-1mm-flatInsert_-_A2B2.m.zip #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 Agilent Quant V8 was used for data analysis #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area normalized MS_METABOLITE_DATA_START Samples S00034622 S00034623 S00034624 S00034625 S00034626 S00034627 S00034628 S00034629 S00034630 S00034631 S00034632 S00034633 S00034634 S00034635 S00034636 S00034637 S00034638 S00034639 S00034640 S00034641 S00034642 S00034643 S00034644 S00034645 S00034646 S00034647 S00034648 S00034649 S00034650 S00034651 Factors Replication:#1 | Treatment:Ctrl Replication:#1 | Treatment:Ctrl Replication:#1 | Treatment:Ctrl Replication:#1 | Treatment:SuccinateConsumingBacteria Replication:#1 | Treatment:SuccinateConsumingBacteria Replication:#2 | Treatment:Ctrl Replication:#2 | Treatment:Ctrl Replication:#2 | Treatment:Ctrl Replication:#2 | Treatment:Ctrl Replication:#2 | Treatment:Ctrl Replication:#2 | Treatment:SuccinateConsumingBacteria Replication:#2 | Treatment:SuccinateConsumingBacteria Replication:#2 | Treatment:SuccinateConsumingBacteria Replication:#2 | Treatment:SuccinateConsumingBacteria Replication:#2 | Treatment:SuccinateConsumingBacteria Replication:#3 | Treatment:Ctrl Replication:#3 | Treatment:Ctrl Replication:#3 | Treatment:Ctrl Replication:#3 | Treatment:Ctrl Replication:#3 | Treatment:Ctrl Replication:#3 | Treatment:Ctrl Replication:#3 | Treatment:Ctrl Replication:#3 | Treatment:SuccinateConsumingBacteria Replication:#3 | Treatment:SuccinateConsumingBacteria Replication:#3 | Treatment:SuccinateConsumingBacteria Replication:#3 | Treatment:SuccinateConsumingBacteria Replication:#3 | Treatment:SuccinateConsumingBacteria Replication:#3 | Treatment:SuccinateConsumingBacteria Replication:#3 | Treatment:SuccinateConsumingBacteria Replication:#3 | Treatment:SuccinateConsumingBacteria STEARIC ACID 198345518 183116852 140240168 234455815 194128103 237120674 284712690 268690828 252873052 291647876 344062199 209840663 236940338 209316610 233439831 197776481 238284059 191918697 134382245 208358132 205445682 190292847 245961911 264987066 218492110 148169360 118875934 212548882 149897994 147915551 OLEIC ACID 418701428 398256327 319180059 491220037 383804209 387914108 478431158 406448667 414173010 490586951 511812654 358736311 436511837 332531035 364606093 381641164 365297528 374288684 271767250 390076896 356553000 324002517 360043595 389861078 328918318 247676740 203744089 349542612 258687565 303737177 PALMITIC ACID 249990853 247127596 188568482 284494675 264840551 274232757 317618255 299081913 277355753 317721355 346176035 258702849 278828017 234410515 272000064 244664916 298742857 251279409 170940373 281749592 251145284 245268919 285429133 283257804 272545336 194448935 153254839 244864095 193885493 196474608 TAURINE 9366535 7284149 11365623 8554694 8820796 6105660 7563650 8214745 7303694 6792147 13598546 13043335 15745267 9458928 5967214 31273646 13380590 17102684 20644912 17054478 32139491 10960365 12738879 10876095 17138947 9288535 5438632 9134353 10704306 10158336 MALATE 71621 76518 57217 32694 69128 55962 31547 34741 33612 53851 45494 70678 48456 72295 47410 55329 55902 58918 64819 43230 41131 37716 39638 21290 35763 64847 46028 43804 56267 57424 ALPHA-KETOGLUTARATE 323254 333042 656168 824512 729901 333971 419870 548742 681152 344366 660254 1318758 525151 329128 571147 1555984 469320 682421 1414988 594899 655473 546326 394782 583390 507155 1186406 586261 609644 979137 815542 CITRATE/ISOCITRATE 170722 273808 242034 172516 474121 148478 235951 278797 98090 280695 250816 171952 132093 422750 165452 181585 216723 420899 410023 274949 188647 212825 178447 292776 303479 263371 204196 280966 157187 176511 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name quantified m/z retention index PubChem ID KEGG ID other_id other_id_type id_confidence ri_type inchi_key STEARIC ACID 283.264 3.386 5281 HMDB0000827 HMDB 1 Time, min QIQXTHQIDYTFRH-UHFFFAOYSA-N OLEIC ACID 281.248 3.469 445639 HMDB0000207 HMDB 1 Time, min ZQPPMHVWECSIRJ-KTKRTIGZSA-N PALMITIC ACID 255.233 3.618 985 HMDB0000220 HMDB 1 Time, min IPCSVZSSVZVIGE-UHFFFAOYSA-N TAURINE 124.0068 8.194 1123 HMDB0000251 HMDB 1 Time, min XOAAWQZATWQOTB-UHFFFAOYSA-N MALATE 133.0137 14.5 525 HMDB0000744 HMDB 1 Time, min BJEPYKJPYRNKOW-UHFFFAOYSA-N ALPHA-KETOGLUTARATE 145.0137 14.611 51 HMDB0000208 HMDB 1 Time, min KPGXRSRHYNQIFN-UHFFFAOYSA-N CITRATE/ISOCITRATE 191.0192 15.4 311 HMDB0000094 HMDB 1 Time, min KRKNYBCHXYNGOX-UHFFFAOYSA-N METABOLITES_END #END