#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