#METABOLOMICS WORKBENCH hormel101_20170623_080927 DATATRACK_ID:1075 STUDY_ID:ST000628 ANALYSIS_ID:AN000960 PROJECT_ID:PR000457 VERSION 1 CREATED_ON June 25, 2017, 8:27 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 TCA Cycle Metabolites of Dietary Salt Effects on Blood Pressure in Human Urine ST:STUDY_TITLE from 20 Participants of the DASH2 Clinical Trial ST:STUDY_SUMMARY The objective of the study is to identify changes of urinary metabolite profiles ST:STUDY_SUMMARY associated with different responses to blood pressure to salt. Subjects are ST:STUDY_SUMMARY derived from The Dietary approaches to stop hypertension (DASH) diet, Sodium ST:STUDY_SUMMARY Intake and Blood Pressure Trial (Sacks FM et al PMID: 11136953,N Engl J Med. ST:STUDY_SUMMARY 2001).We choose two groups subjects who meet the following conditions(the two ST:STUDY_SUMMARY groups are separately named A and B). We chose subjects on the Control diet ST:STUDY_SUMMARY .These subjects meet the blood pressure criteria described below:Group A ST:STUDY_SUMMARY subjects conditions: 1) On Control diet. 2) Normotensive subjects: systolic ST:STUDY_SUMMARY blood pressure from the low sodium visit is less than 140 and the diastolic ST:STUDY_SUMMARY blood pressure from low sodium visit is less than 90; 3) For group A: Either the ST:STUDY_SUMMARY systolic blood pressure from the high sodium visit was greater than 10 mmHg ST:STUDY_SUMMARY higher than the systolic blood pressure from the low sodium visit, or the ST:STUDY_SUMMARY diastolic blood pressure from the high sodium visit was greater than 10 mmHg ST:STUDY_SUMMARY higher than the diastolic blood pressure from the low sodium visit; 3) For group ST:STUDY_SUMMARY B: The systolic blood pressure from the high sodium visit is within 5 mmHg (i.e. ST:STUDY_SUMMARY +/- 5) from the systolic blood pressure from the low sodium visit, and the ST:STUDY_SUMMARY diastolic blood pressure from the high sodium visit is within 5 mmHg from the ST:STUDY_SUMMARY diastolic blood pressure from the low sodium visit. Use gas chromatography/mass ST:STUDY_SUMMARY spectrometry (GC/MS) analysis, and liquid chromatography/mass spectrometry ST:STUDY_SUMMARY (LC/MS)analysis to find the differences of metabolic profiles between the high ST:STUDY_SUMMARY sodium level and the low sodium level, and compare the metabolic profiles of A ST:STUDY_SUMMARY with the metabolic profiles of B at the low and high sodium level. Note, this ST:STUDY_SUMMARY was used as a pilot, the rest of the samples are in another uploaded study. 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 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 DASH200367 Sample #1 Sodium level:H | grouping:A(salt sensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200367 Sample #2 Sodium level:L | grouping:A(salt sensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200298 Sample #3 Sodium level:L | grouping:A(salt sensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200298 Sample #4 Sodium level:H | grouping:A(salt sensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200052 Sample #5 Sodium level:H | grouping:A(salt sensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200052 Sample #6 Sodium level:L | grouping:A(salt sensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200136 Sample #7 Sodium level:L | grouping:A(salt sensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200136 Sample #8 Sodium level:H | grouping:A(salt sensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200035 Sample #9 Sodium level:H | grouping:A(salt sensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200035 Sample #10 Sodium level:L | grouping:A(salt sensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200125 Sample #21 Sodium level:L | grouping:B(salt insensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200125 Sample #22 Sodium level:H | grouping:B(salt insensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200161 Sample #23 Sodium level:H | grouping:B(salt insensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200161 Sample #24 Sodium level:L | grouping:B(salt insensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200218 Sample #25 Sodium level:H | grouping:B(salt insensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200218 Sample #26 Sodium level:L | grouping:B(salt insensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200238 Sample #27 Sodium level:L | grouping:B(salt insensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200238 Sample #28 Sodium level:H | grouping:B(salt insensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200167 Sample #29 Sodium level:L | grouping:B(salt insensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200167 Sample #30 Sodium level:H | grouping:B(salt insensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200050 Sample #11 Sodium level:H | grouping:A(salt sensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200050 Sample #12 Sodium level:L | grouping:A(salt sensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200064 Sample #13 Sodium level:H | grouping:A(salt sensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200064 Sample #14 Sodium level:L | grouping:A(salt sensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200100 Sample #15 Sodium level:H | grouping:A(salt sensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200100 Sample #16 Sodium level:L | grouping:A(salt sensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200375 Sample #17 Sodium level:H | grouping:A(salt sensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200375 Sample #18 Sodium level:L | grouping:A(salt sensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200060 Sample #19 Sodium level:H | grouping:A(salt sensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200060 Sample #20 Sodium level:L | grouping:A(salt sensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200199 Sample #31 Sodium level:L | grouping:B(salt insensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200199 Sample #32 Sodium level:H | grouping:B(salt insensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200306 Sample #33 Sodium level:H | grouping:B(salt insensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200306 Sample #34 Sodium level:L | grouping:B(salt insensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200193 Sample #35 Sodium level:H | grouping:B(salt insensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200193 Sample #36 Sodium level:L | grouping:B(salt insensitive) gender=1 SUBJECT_SAMPLE_FACTORS DASH200244 Sample #37 Sodium level:H | grouping:B(salt insensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200244 Sample #38 Sodium level:L | grouping:B(salt insensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200389 Sample #39 Sodium level:H | grouping:B(salt insensitive) gender=2 SUBJECT_SAMPLE_FACTORS DASH200389 Sample #40 Sodium level:L | grouping:B(salt insensitive) gender=2 #COLLECTION CO:COLLECTION_SUMMARY The urine samples we have were 24-hour collections occurring in the fourth week CO:COLLECTION_SUMMARY on either the low- or the high-sodium intake level. In total, 120 human samples CO:COLLECTION_SUMMARY (60 subjects, a low-sodium sample and a high-sodium sample for each subject) CO:COLLECTION_SUMMARY will be analyzed. #TREATMENT TR:TREATMENT_SUMMARY We will analyze urine specimens collected during the Dietary Approaches to Stop TR:TREATMENT_SUMMARY Hypertension – Sodium Study (DASH-Sodium or DASH2) clinical trial. More than TR:TREATMENT_SUMMARY 400 participants in the study were divided into two groups. One followed a TR:TREATMENT_SUMMARY common Western diet, while the other group followed the “healthier” DASH TR:TREATMENT_SUMMARY diet. Then each group had three 30 day periods, in random orders, that TR:TREATMENT_SUMMARY corresponded to high, intermediate, or low sodium intakes while eating their TR:TREATMENT_SUMMARY designated diet. The participants had all of their meals prepared and provided TR:TREATMENT_SUMMARY for them during the entire study. It was found that reducing sodium intake TR:TREATMENT_SUMMARY significantly reduced blood pressure especially when on the common Western diet. TR:TREATMENT_SUMMARY We have obtained urine samples and phenotypic data for a subset of the TR:TREATMENT_SUMMARY participants of the DASH2 trial form the NHLBI biospecimen and data repository TR:TREATMENT_SUMMARY BioLINCC. We selected two sub-groups of participants on the Western diet. Group TR:TREATMENT_SUMMARY A (n=31) exhibited large (>12 mmHg) differences in systolic or diastolic TR:TREATMENT_SUMMARY pressure between low- and high-sodium intakes (“salt-sensitive” group). TR:TREATMENT_SUMMARY Group B (n=29) showed similar blood pressure (differences <5 mmHg for both TR:TREATMENT_SUMMARY systolic and diastolic pressures) between low- and high-sodium intakes TR:TREATMENT_SUMMARY (“saltinsensitive” group). #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 2 drops of 6 N HCl was added in every 8 ml urine samples when the 24 hour urine SP:SAMPLEPREP_SUMMARY was collected. All urine samples were stored in -80℃ freezer. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 7890B CH:COLUMN_NAME Agilent HP5-MS (30m × 0.25mm, 0.25 um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 5977A MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS uM MS_METABOLITE_DATA_START Samples Sample #1 Sample #2 Sample #3 Sample #4 Sample #5 Sample #6 Sample #7 Sample #8 Sample #9 Sample #10 Sample #21 Sample #22 Sample #23 Sample #24 Sample #25 Sample #26 Sample #27 Sample #28 Sample #29 Sample #30 Sample #11 Sample #12 Sample #13 Sample #14 Sample #15 Sample #16 Sample #17 Sample #18 Sample #19 Sample #20 Sample #31 Sample #32 Sample #33 Sample #34 Sample #35 Sample #36 Sample #37 Sample #38 Sample #39 Sample #40 Factors Sodium level:H | grouping:A(salt sensitive) Sodium level:L | grouping:A(salt sensitive) Sodium level:L | grouping:A(salt sensitive) Sodium level:H | grouping:A(salt sensitive) Sodium level:H | grouping:A(salt sensitive) Sodium level:L | grouping:A(salt sensitive) Sodium level:L | grouping:A(salt sensitive) Sodium level:H | grouping:A(salt sensitive) Sodium level:H | grouping:A(salt sensitive) Sodium level:L | grouping:A(salt sensitive) Sodium level:L | grouping:B(salt insensitive) Sodium level:H | grouping:B(salt insensitive) Sodium level:H | grouping:B(salt insensitive) Sodium level:L | grouping:B(salt insensitive) Sodium level:H | grouping:B(salt insensitive) Sodium level:L | grouping:B(salt insensitive) Sodium level:L | grouping:B(salt insensitive) Sodium level:H | grouping:B(salt insensitive) Sodium level:L | grouping:B(salt insensitive) Sodium level:H | grouping:B(salt insensitive) Sodium level:H | grouping:A(salt sensitive) Sodium level:L | grouping:A(salt sensitive) Sodium level:H | grouping:A(salt sensitive) Sodium level:L | grouping:A(salt sensitive) Sodium level:H | grouping:A(salt sensitive) Sodium level:L | grouping:A(salt sensitive) Sodium level:H | grouping:A(salt sensitive) Sodium level:L | grouping:A(salt sensitive) Sodium level:H | grouping:A(salt sensitive) Sodium level:L | grouping:A(salt sensitive) Sodium level:L | grouping:B(salt insensitive) Sodium level:H | grouping:B(salt insensitive) Sodium level:H | grouping:B(salt insensitive) Sodium level:L | grouping:B(salt insensitive) Sodium level:H | grouping:B(salt insensitive) Sodium level:L | grouping:B(salt insensitive) Sodium level:H | grouping:B(salt insensitive) Sodium level:L | grouping:B(salt insensitive) Sodium level:H | grouping:B(salt insensitive) Sodium level:L | grouping:B(salt insensitive) Lactate 102.5 96.8 80.8 60.4 67.6 76.9 2010.9 174.3 65.9 98.5 82.8 197.1 146.4 145.6 180.5 172.2 88.0 82.8 235.1 175.0 67.8 121.4 45.4 100.5 79.4 65.0 253.9 202.7 90.6 86.0 33.5 24.9 72.9 84.0 346.2 74.8 496.2 451.9 212.7 273.8 Succinate 30.1 17.0 21.4 13.9 6.9 10.1 78.9 57.1 15.2 19.3 15.5 860.0 42.2 38.6 33.2 1126.5 13.7 10.2 218.9 194.2 15.8 19.4 64.0 37.4 51.8 14.4 10.9 10.3 18.8 15.7 4.6 20.1 24.4 16.3 6.4 5.4 198.7 217.2 18.2 29.2 Fumarate 8.7 7.1 7.1 8.0 5.6 7.7 17.4 9.2 8.1 8.1 5.3 21.7 11.5 10.7 7.4 8.9 5.5 4.1 10.9 8.2 6.0 5.9 4.8 6.1 8.7 6.0 6.1 6.6 8.5 7.8 2.8 2.9 11.9 8.8 7.9 3.6 3.4 3.7 6.2 8.7 Oxaloacetate 3.5 3.5 3.0 2.0 1.0 1.6 1.8 1.4 1.7 2.4 1.9 0.9 0.9 0.9 2.1 0.5 3.0 0.7 1.3 1.9 2.4 4.7 1.1 1.5 1.7 0.9 0.9 0.9 1.9 1.1 0.7 0.4 2.0 1.3 0.6 0.4 1.6 0.5 1.4 2.0 Ketoglutarate 128.3 87.9 64.7 32.6 17.5 35.9 316.2 79.3 59.1 78.2 65.3 46.8 62.2 92.2 117.6 15.4 18.9 27.7 62.0 50.8 22.0 59.0 42.8 55.6 89.0 68.5 44.0 25.6 101.3 94.0 13.6 11.7 20.9 26.8 41.5 12.5 167.0 158.6 61.5 80.4 Malate 9.0 6.5 5.4 4.2 4.4 4.9 29.2 13.6 14.2 12.5 5.0 375.1 14.0 16.4 7.6 6.8 130.0 42.3 15.3 13.1 7.5 6.5 6.4 8.0 5.4 6.1 8.3 9.2 8.1 7.9 3.2 1.6 10.4 7.4 12.1 2.4 18.6 33.3 7.7 11.7 cis-Aconitic Acid 280.9 202.7 166.1 150.1 101.1 169.6 200.6 107.7 72.7 104.7 130.3 289.0 79.4 99.1 139.7 264.7 96.4 99.7 170.1 114.0 147.8 318.6 109.4 123.5 169.4 207.1 87.8 81.6 168.4 159.5 59.6 98.8 204.9 149.2 101.2 72.3 100.5 110.1 104.9 152.4 Citrate 2551.1 2186.5 1152.6 759.6 263.6 323.6 4929.8 3288.9 2483.1 4145.6 2447.3 237.0 1852.8 2265.8 2589.8 5043.9 524.4 595.8 719.7 964.6 1103.0 768.0 1587.4 1849.0 1515.0 464.9 1676.2 1799.3 2693.2 2629.6 702.5 425.6 1550.9 1114.8 902.6 532.3 2408.4 2020.9 1713.5 2479.4 Isocitrate 374.1 273.3 235.5 209.0 141.1 236.2 228.6 140.1 115.6 165.2 181.7 164.9 103.0 121.7 223.2 384.4 119.1 117.2 223.7 169.9 183.4 393.7 132.6 163.0 220.6 166.7 105.6 100.8 208.0 198.3 72.8 89.3 259.2 192.7 102.3 80.3 128.9 126.8 126.7 190.9 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Lactate Succinate Fumarate Oxaloacetate Ketoglutarate Malate cis-Aconitic Acid Citrate Isocitrate METABOLITES_END #END