#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