#METABOLOMICS WORKBENCH hormel101_20180705_083725_mwtab.txt DATATRACK_ID:1434 STUDY_ID:ST000996 ANALYSIS_ID:AN001625 PROJECT_ID:PR000675 VERSION 1 CREATED_ON July 5, 2018, 3:11 pm #PROJECT PR:PROJECT_TITLE Mayo Pilot and Feasibility: The Enterohepatic Metabolome in Primary Sclerosing PR:PROJECT_TITLE Cholangitis PR:PROJECT_SUMMARY Emerging in vitro and in vivo data, including work from our laboratory and PR:PROJECT_SUMMARY clinical research group, suggest fundamental pathophysiologic mechanisms in PR:PROJECT_SUMMARY primary sclerosing cholangitis (PSC) that are centered on the enterohepatic PR:PROJECT_SUMMARY circulation of gut-derived molecules. Therefore, in this proposal, we will test PR:PROJECT_SUMMARY the central hypothesis that increased pathologic enterohepatic circulation of PR:PROJECT_SUMMARY enteric metabolites which trigger specific pro-fibroinflammatory hepatobiliary PR:PROJECT_SUMMARY responses are centrally involved in the etiopathogenesis of primary sclerosing PR:PROJECT_SUMMARY cholangitis (PSC). While these processes have been hypothesized to play a PR:PROJECT_SUMMARY significant role in the initiation, progression, and adverse clinical sequelae PR:PROJECT_SUMMARY of PSC, they have not been directly tested to date. In our proposal, we will PR:PROJECT_SUMMARY experimentally address the nature and extent of the metabolomic profiles of PR:PROJECT_SUMMARY portal and peripheral blood as well as bile in patients with PSC. We will PR:PROJECT_SUMMARY perform qualitative and quantitative ultra-performance liquid PR:PROJECT_SUMMARY chromatography/mass spectroscopy-based metabolomic analyses to determine PR:PROJECT_SUMMARY metabolic changes in portal and peripheral plasma and bile. Through subsequent PR:PROJECT_SUMMARY pathway analyses we intend to identify metabolic enzymes and known biochemical PR:PROJECT_SUMMARY pathways that may be altered in PSC. We anticipate that patients with PSC will PR:PROJECT_SUMMARY have distinct alterations in the portal venous and bile metabolomic profiles and PR:PROJECT_SUMMARY associated signaling pathways compared to normal and disease controls; and these PR:PROJECT_SUMMARY alterations may be amenable to pharmacologic manipulation and future therapies. PR:INSTITUTE Mayo Clinic PR:LAST_NAME O'Hara PR:FIRST_NAME Steven PR:ADDRESS 200 First St. SW, Rochester, Minnesota, 55905, USA PR:EMAIL ohara.steven@mayo.edu PR:PHONE 507-284-1006 #STUDY ST:STUDY_TITLE Non-Esterified Fatty Acids of Primary Sclerosing Cholangitis (part II) ST:STUDY_SUMMARY To qualitatively and quantitatively analyze enterohepatically-circulated ST:STUDY_SUMMARY molecules using targeted free fatty acid concentrations of peripheral blood ST:STUDY_SUMMARY collected from primary sclerosing cholangitis (PSC) patients compared to normal ST:STUDY_SUMMARY and diseased controls. There are three groups of patients. (1) Normal donor ST:STUDY_SUMMARY controls (ND), (2) Patients with Primary Sclerosing Cholangitis (PSC), and (3) ST:STUDY_SUMMARY Disease Controls (DC) which are patients with liver disease other than PSC. ST:INSTITUTE Mayo Clinic ST:LAST_NAME O'Hara ST:FIRST_NAME Steven ST:ADDRESS 200 First St. SW, Rochester, Minnesota, 55905, USA ST:EMAIL ohara.steven@mayo.edu ST:PHONE 507-284-1006 #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 - ms6722-1 Grouping:ND Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-2 Grouping:ND Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-3 Grouping:ND Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-4 Grouping:ND Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-5 Grouping:ND Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-6 Grouping:ND Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-7 Grouping:ND Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-8 Grouping:ND Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-9 Grouping:ND Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-10 Grouping:PSC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-11 Grouping:PSC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-12 Grouping:PSC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-13 Grouping:PSC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-14 Grouping:PSC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-15 Grouping:PSC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-16 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-17 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-18 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-19 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-20 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-21 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-22 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-23 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-24 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-25 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-26 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-27 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-28 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-29 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-30 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-31 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-32 Grouping:DC Sample type=serum; Species=Human SUBJECT_SAMPLE_FACTORS - ms6722-33 Grouping:DC Sample type=serum; Species=Human #COLLECTION CO:COLLECTION_SUMMARY After obtaining informed consent, portal and peripheral venous blood (4 ml of CO:COLLECTION_SUMMARY each) and bile (2 mL) was collected intraoperatively in a red-top tube by a Mayo CO:COLLECTION_SUMMARY Clinic LT surgeon. Blood was placed on ice, promptly fractionated by CO:COLLECTION_SUMMARY centrifugation, divided into 100 μL aliquots, and stored at -80°C. CO:SAMPLE_TYPE Blood (serum) #TREATMENT TR:TREATMENT_SUMMARY " We prospectively enrolled three groups of participants from the Mayo Clinic TR:TREATMENT_SUMMARY Liver Transplant inpatient service and outpatient clinics and have collected TR:TREATMENT_SUMMARY samples from: i) 9 patients with PSC who underwent living- donor LT, ii) 15 TR:TREATMENT_SUMMARY donors (normal controls), and iii) 20 patients with cirrhosis due to a disorder TR:TREATMENT_SUMMARY other than PSC who underwent LT (disease controls). The following inclusion and TR:TREATMENT_SUMMARY exclusion criteria were applied: Inclusion criteria 1. Adult (age>18 years). 2. TR:TREATMENT_SUMMARY PSC patient undergoing LT, healthy living donor, or other chronic liver disease TR:TREATMENT_SUMMARY patient undergoing LT. Exclusion Criteria: 1. Females who are pregnant or TR:TREATMENT_SUMMARY attempting to become pregnant. 2. Concomitant liver disease (e.g. chronic viral TR:TREATMENT_SUMMARY hepatitis in addition to PSC). 3. Acute intestinal disease (infectious TR:TREATMENT_SUMMARY enterocolitis, IBD flare) in the past 6 months. 4. Treatment with any TR:TREATMENT_SUMMARY investigational drugs within the past 6 months. 5. Use of antibiotics within the TR:TREATMENT_SUMMARY past 4 weeks. 6. Any previous organ transplant. 7. Hemodialysis." #SAMPLEPREP SP:SAMPLEPREP_SUMMARY nefa acid concentrations #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1290 Infinity CH:COLUMN_NAME Waters Acquity BEH C18 (150 x 2.1mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6460 QQQ MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS uM #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS uM MS_METABOLITE_DATA_START Samples ms6722-1 ms6722-2 ms6722-3 ms6722-4 ms6722-5 ms6722-6 ms6722-7 ms6722-8 ms6722-9 ms6722-10 ms6722-11 ms6722-12 ms6722-13 ms6722-14 ms6722-15 ms6722-16 ms6722-17 ms6722-18 ms6722-19 ms6722-20 ms6722-21 ms6722-22 ms6722-23 ms6722-24 ms6722-25 ms6722-26 ms6722-27 ms6722-28 ms6722-29 ms6722-30 ms6722-31 ms6722-32 ms6722-33 Factors Grouping:ND Grouping:ND Grouping:ND Grouping:ND Grouping:ND Grouping:ND Grouping:ND Grouping:ND Grouping:ND Grouping:PSC Grouping:PSC Grouping:PSC Grouping:PSC Grouping:PSC Grouping:PSC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC Grouping:DC EPA 0.4 1.2 1.1 0.3 0.7 0.4 0.4 0.4 0.4 0.3 0.6 0.2 0.6 0.4 0.2 0.3 0.4 0.3 0.3 0.4 0.1 0.3 0.7 0.4 0.4 0.2 0.2 0.3 0.3 0.5 0.2 0.3 0.7 linolenic 30.8 24.8 27.9 15.7 20.6 23.3 16.5 25.6 15.6 23.2 44.3 14.8 47.1 32.0 8.1 26.5 55.0 30.0 40.7 46.8 5.2 44.2 7.7 33.3 36.3 18.4 27.1 5.9 15.4 58.0 37.2 2.1 32.6 DHA 4.5 3.6 3.4 2.3 2.7 2.7 2.7 3.1 1.9 2.9 4.6 1.7 4.6 3.1 0.8 2.9 3.2 3.3 2.1 3.5 0.9 2.5 5.4 3.3 3.3 4.2 2.5 2.3 2.8 3.7 1.8 1.8 7.9 myristic 16.1 13.1 20.7 9.9 24.9 23.6 16.1 24.9 11.0 17.6 46.7 12.6 49.2 44.2 4.8 18.8 38.8 42.2 31.2 28.4 10.9 36.2 8.0 36.6 38.5 19.0 28.6 5.2 19.9 33.0 31.1 4.0 28.9 palmitoleic 20.2 19.6 40.3 28.0 46.1 27.8 28.3 62.7 16.5 40.7 134.3 14.4 134.8 187.6 8.1 47.6 192.2 143.1 68.8 63.5 65.1 131.8 23.8 143.3 147.6 66.4 56.9 10.8 47.8 187.3 60.0 11.6 57.7 arachidonic 7.0 9.1 9.0 5.7 5.8 5.5 5.1 6.9 4.8 4.9 5.6 2.7 5.7 3.6 5.2 4.9 5.5 4.9 4.3 5.4 2.3 5.2 4.4 4.9 5.1 4.3 3.7 3.2 3.7 4.8 4.4 5.7 6.1 palmitelaidic 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 linoleic 225.1 167.8 218.8 206.9 172.5 200.2 131.0 227.8 137.2 204.8 272.0 106.9 268.5 157.1 75.0 207.8 350.3 234.4 255.8 332.3 82.4 235.6 59.2 232.3 246.6 147.5 208.8 62.5 161.7 299.9 309.1 44.7 205.9 palmitic 280.3 195.6 308.7 175.2 289.8 251.8 190.4 300.8 115.3 235.8 377.0 159.6 402.3 298.0 116.5 247.9 484.6 333.2 321.9 325.8 141.4 341.2 106.8 384.9 389.3 195.8 282.1 87.3 272.8 348.8 387.9 88.5 341.4 oleic 366 219 491 279 412 400 292 444 168 409 600 229 640 488 115 445 1004 586 516 501 297 560 154 630 662 387 444 145 376 661 655 67 576 elaidic 14.1 9.2 16.1 10.9 15.7 16.3 16.3 21.1 4.4 9.3 24.8 13.1 21.2 9.5 7.2 12.6 25.8 16.9 19.1 27.1 8.9 17.6 7.6 20.1 23.1 9.4 21.3 8.6 21.2 26.2 20.1 4.3 16.8 stearic 57.0 47.8 55.7 32.5 51.4 59.0 49.9 47.2 30.0 40.0 47.0 49.1 47.6 22.7 30.8 42.6 40.7 29.1 40.4 47.7 26.3 46.9 19.4 44.2 43.3 34.7 73.4 52.6 50.7 30.0 82.6 14.4 74.5 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name EPA linolenic DHA myristic palmitoleic arachidonic palmitelaidic linoleic palmitic oleic elaidic stearic METABOLITES_END #END