#METABOLOMICS WORKBENCH araskind_20191211_122141 DATATRACK_ID:1866 STUDY_ID:ST001287 ANALYSIS_ID:AN002147 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 pmol/mg MS_METABOLITE_DATA_START Samples S00034622 S00034631 S00034632 S00034633 S00034634 S00034635 S00034636 S00034637 S00034638 S00034639 S00034640 S00034623 S00034641 S00034642 S00034643 S00034644 S00034645 S00034646 S00034647 S00034648 S00034649 S00034650 S00034624 S00034651 S00034625 S00034626 S00034627 S00034628 S00034629 S00034630 Factors Replication:#1 | 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:#1 | 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:#1 | Treatment:Ctrl Replication:#3 | Treatment:SuccinateConsumingBacteria Replication:#1 | Treatment:SuccinateConsumingBacteria Replication:#1 | Treatment:SuccinateConsumingBacteria Replication:#2 | Treatment:Ctrl Replication:#2 | Treatment:Ctrl Replication:#2 | Treatment:Ctrl Replication:#2 | Treatment:Ctrl SUCCINIC ACID 343.63 605.50 352.73 229.49 250.39 306.44 197.47 404.17 422.75 229.54 307.26 294.50 430.40 340.53 327.97 121.54 242.78 132.50 296.97 205.91 283.22 268.98 389.46 545.91 128.83 150.09 380.83 395.79 589.89 458.74 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 SUCCINIC ACID 117.0188 14.46 1110 HMDB0000254 HMDB 1 Time, min KDYFGRWQOYBRFD-UHFFFAOYSA-N METABOLITES_END #END