#METABOLOMICS WORKBENCH hormel101_20170623_110816 DATATRACK_ID:1081 STUDY_ID:ST000633 ANALYSIS_ID:AN000965 PROJECT_ID:PR000457
VERSION             	1
CREATED_ON             	June 25, 2017, 9:07 am
#PROJECT
PR:PROJECT_TITLE                 	Metabolomic Mechanisms of Dietary Salt Effects on Blood Pressure
PR:PROJECT_SUMMARY               	"Enhanced sensitivity of blood pressure to salt intake is observed in
PR:PROJECT_SUMMARY               	approximately 50% of hypertensive patients, reaching 75% in African American
PR:PROJECT_SUMMARY               	hypertensive patients. We recently discovered a novel role of abnormal cellular
PR:PROJECT_SUMMARY               	intermediary metabolism in hypertension in the Dahl salt-sensitive (SS) rat, the
PR:PROJECT_SUMMARY               	most commonly used polygenic, hereditary model of human salt-sensitive
PR:PROJECT_SUMMARY               	hypertension. We propose to test the hypothesis that blood pressure sensitivity
PR:PROJECT_SUMMARY               	to dietary salt intake in human is associated with metabolite changes in the
PR:PROJECT_SUMMARY               	urine. Leveraging the expertise and resources at the Mayo Clinic Metabolomics
PR:PROJECT_SUMMARY               	Resources Core, we propose to perform targeted LC/MS analysis and NMR spectra
PR:PROJECT_SUMMARY               	generation in urine samples obtained from a subset of subjects from the Dietary
PR:PROJECT_SUMMARY               	Approaches to Stop Hypertension – Sodium (DASH2) clinical trial and kidney
PR:PROJECT_SUMMARY               	tissue extract and urine samples from SS rats and a newly generated transgenic
PR:PROJECT_SUMMARY               	rat that overexpresses fumarase (SS.Fh1+). The study will be the first to
PR:PROJECT_SUMMARY               	systematically characterize urinary metabolite profiles associated with blood
PR:PROJECT_SUMMARY               	pressure response to salt in humans. The study is anticipated to generate new
PR:PROJECT_SUMMARY               	insight into the mechanisms (particularly renal mechanisms) underlying
PR:PROJECT_SUMMARY               	salt-sensitive hypertension. Findings of the proposed study could lead to an
PR:PROJECT_SUMMARY               	expanded clinical study as well as mechanistic studies in animal models."
PR:INSTITUTE                     	Mayo Clinic
PR:LAST_NAME                     	Liang
PR:FIRST_NAME                    	Mingyu
PR:ADDRESS                       	Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
PR:EMAIL                         	mliang@mcw.edu
PR:PHONE                         	414-955-8539
#STUDY
ST:STUDY_TITLE                   	Neuromodulator Metabolites of Dietary Salt Effects on Blood Pressure in Rat
ST:STUDY_TITLE                   	Urine (part VI)
ST:STUDY_SUMMARY                 	We propose to analyze kidney tissue extract and urine samples from SS and
ST:STUDY_SUMMARY                 	SS.Fh1+ transgenic rats in addition to the analysis of urine samples from the
ST:STUDY_SUMMARY                 	DASH2 trial. The analysis of the rat samples will be highly valuable for several
ST:STUDY_SUMMARY                 	reasons. First, it will to take the findings in human subjects back to animal
ST:STUDY_SUMMARY                 	models and prepare us for further mechanistic studies. We hypothesize at least
ST:STUDY_SUMMARY                 	some of the effects of dietary salt intake on metabolite profiles in human will
ST:STUDY_SUMMARY                 	be recapitulated or altered in the SS rat. If this is confirmed, we will have a
ST:STUDY_SUMMARY                 	highly informative animal model ready for mechanistic studies in which we can
ST:STUDY_SUMMARY                 	investigate the functional contribution of specific metabolites to hypertension
ST:STUDY_SUMMARY                 	and the mechanisms involved. Second, the rat study will allow us to take
ST:STUDY_SUMMARY                 	advantage of a new and unique transgenic SS.Fh1+ model that we recently
ST:STUDY_SUMMARY                 	developed that overexpresses fumarase (Fh1) on the genetic background of the SS
ST:STUDY_SUMMARY                 	rat. Fumarase is a TCA cycle enzyme previously implicated in salt-induced
ST:STUDY_SUMMARY                 	hypertension in SS rats.
ST:INSTITUTE                     	Mayo Clinic
ST:LAST_NAME                     	Liang
ST:FIRST_NAME                    	Mingyu
ST:ADDRESS                       	Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
ST:EMAIL                         	mliang@mcw.edu
ST:PHONE                         	414-955-8539
#SUBJECT
SU:SUBJECT_TYPE                  	Rat
SU:SUBJECT_SPECIES               	Rattus norvegicus
SU:TAXONOMY_ID                   	10116
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	ms5954-1	Time point:Control	Date of study=8/3/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-2	Time point:Control	Date of study=8/3/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-3	Time point:Control	Date of study=8/3/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-4	Time point:Control	Date of study=8/3/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-5	Time point:Control	Date of study=8/3/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-6	Time point:Control	Date of study=8/3/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-7	Time point:Control	Date of study=8/3/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-8	Time point:Control	Date of study=8/3/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-9	Time point:7 day HS	Date of study=8/10/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-10	Time point:7 day HS	Date of study=8/10/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-11	Time point:7 day HS	Date of study=8/10/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-12	Time point:7 day HS	Date of study=8/10/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-13	Time point:7 day HS	Date of study=8/10/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-14	Time point:7 day HS	Date of study=8/10/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-15	Time point:7 day HS	Date of study=8/10/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-16	Time point:7 day HS	Date of study=8/10/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-17	Time point:Control	Date of study=9/2/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-18	Time point:Control	Date of study=9/2/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-19	Time point:Control	Date of study=9/2/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-20	Time point:Control	Date of study=9/2/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-21	Time point:Control	Date of study=9/2/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-22	Time point:Control	Date of study=9/2/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-23	Time point:Control	Date of study=9/2/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-24	Time point:Control	Date of study=9/2/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-25	Time point:Control	Date of study=9/2/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-26	Time point:Control	Date of study=9/2/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-27	Time point:7 day HS	Date of study=9/9/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-28	Time point:7 day HS	Date of study=9/9/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-29	Time point:7 day HS	Date of study=9/9/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-30	Time point:7 day HS	Date of study=9/9/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-31	Time point:7 day HS	Date of study=9/9/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-32	Time point:7 day HS	Date of study=9/9/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-33	Time point:7 day HS	Date of study=9/9/2010; gender=M; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-34	Time point:7 day HS	Date of study=9/9/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-35	Time point:7 day HS	Date of study=9/9/2010; gender=F; sample type=rat urine
SUBJECT_SAMPLE_FACTORS           	-	ms5954-36	Time point:7 day HS	Date of study=9/9/2010; gender=F; sample type=rat urine
#COLLECTION
CO:COLLECTION_SUMMARY            	Samples are from Transgenic fumerase rats and WT littermates, 13 weeks of age,
CO:COLLECTION_SUMMARY            	placed on low salt or high salt dyets. We are interested in any differences in
CO:COLLECTION_SUMMARY            	the TCA and AA intermediates.
#TREATMENT
TR:TREATMENT_SUMMARY             	We will analyze kidney tissue extract and urine samples from SS rats and the
TR:TREATMENT_SUMMARY             	newly generated SS.Fh1+ transgenic rats. The SS.Fh1+ rat was generated using a
TR:TREATMENT_SUMMARY             	Sleeping Beauty transposon-mediated transgenic technique. Overexpression of
TR:TREATMENT_SUMMARY             	fumarase has been confirmed in SS.Fh1+ rats. Preliminary study indicated that
TR:TREATMENT_SUMMARY             	the development of salt-induced hypertension was altered in SS.Fh1+ rats
TR:TREATMENT_SUMMARY             	compared to wild-type littermate SS rats. We will analyze kidney tissue extract
TR:TREATMENT_SUMMARY             	and urine samples collected from rats maintained on a 0.4% NaCl diet or switched
TR:TREATMENT_SUMMARY             	to a 4% NaCl diet for 7 days. The dietary protocol has been used in numerous
TR:TREATMENT_SUMMARY             	studies to examine mechanisms underlying salt sensitivity including early
TR:TREATMENT_SUMMARY             	responses (7 days) to salt in the SS model. In total, 64 rat samples (two rat
TR:TREATMENT_SUMMARY             	strains, two salt levels, and 8 rats per condition) will be analyzed.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	rat urine neuromodulators
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Waters Acquity
CH:COLUMN_NAME                   	Waters Acquity BEH C18 (150 x 2.1mm, 1.7um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:MS_COMMENTS                   	-
MS:INSTRUMENT_NAME               	Thermo Quantum Ultra
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	uM
MS_METABOLITE_DATA_START
Samples	ms5954-1	ms5954-2	ms5954-3	ms5954-4	ms5954-5	ms5954-6	ms5954-7	ms5954-8	ms5954-9	ms5954-10	ms5954-11	ms5954-12	ms5954-13	ms5954-14	ms5954-15	ms5954-16	ms5954-17	ms5954-18	ms5954-19	ms5954-20	ms5954-21	ms5954-22	ms5954-23	ms5954-24	ms5954-25	ms5954-26	ms5954-27	ms5954-28	ms5954-29	ms5954-30	ms5954-31	ms5954-32	ms5954-33	ms5954-34	ms5954-35	ms5954-36
Factors	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:Control	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS	Time point:7 day HS
Acetylcholine	67.0	144.7	270.4	73.4	17.1	9.0	34.7	53.5	19.5	13.1	16.1	11.8	2.7	5.4	3.8	2.5	80.2	194.7	7.4	6.9	189.5	88.9	328.3	16.0	8.5	12.4	16.3	19.1	3.3	4.0	25.4	21.6	14.5	8.6	3.1	5.2
Adenosine	109.9	667.6	258.9	59.8	119.4	9.5	82.5	692.8	6.1	13.7	3.3	5.4	4.6	9.6	42.7	3.6	64.2	25.2	25.7	7.7	195.1	65.3	60.9	23.5	32.2	5.4	3.3	4.1	3.3	9.5	3.3	10.4	9.1	5.2	2.9	35.7
Norepinephrine	1.2	1.9	1.7	1.2	1.7	0.7	1.6	2.3	0.1	0.4	0.4	0.4	0.3	0.3	0.3	0.4	1.9	1.5	1.3	1.0	1.4	1.7	1.4	2.2	1.7	1.1	0.2	0.3	0.5	0.7	0.3	0.3	0.0	0.5	0.6	0.6
Dopamine	8.1	12.2	11.5	3.2	9.6	2.8	14.1	15.7	0.8	0.9	1.5	1.1	1.5	1.3	1.1	1.4	8.3	5.5	4.8	4.1	4.7	3.2	7.9	6.9	4.3	2.5	0.6	0.5	2.3	0.9	1.5	1.0	0.1	1.7	1.8	1.4
Serotonin	15.4	18.6	29.6	9.5	21.0	10.4	39.7	42.1	3.0	5.8	6.2	5.0	5.5	5.8	4.2	10.5	19.8	15.8	16.7	17.2	15.6	14.9	29.6	22.0	14.7	23.1	4.1	3.3	9.3	5.1	7.2	3.7	2.2	9.8	13.2	5.7
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
Acetylcholine
Adenosine
Norepinephrine
Dopamine
Serotonin
METABOLITES_END
#END