{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001917","ANALYSIS_ID":"AN003115","VERSION":"1","CREATED_ON":"September 13, 2021, 5:34 pm"},

"PROJECT":{"PROJECT_TITLE":"Myocardial Rev-erb-mediated diurnal metabolic rhythm","PROJECT_SUMMARY":"Agonists and antagonists of nuclear receptor Rev-erbα/β, key components of the circadian clock, can benefit the heart. Here, we show that mice with cardiomyocyte-specific knockout (KO) of Rev-erbα/β display progressive cardiac dilation and lethal heart failure. Inducible ablation of Rev-erbα/β in adult hearts causes similar phenotypes. Impaired fatty acid oxidation in the KO myocardium, particularly in the light cycle, precedes contractile dysfunctions with a reciprocal overreliance on carbohydrate utilization, particularly in the dark cycle. These findings delineate temporal coordination between clock-mediated anticipation and nutrient-induced response in myocardial metabolism.","INSTITUTE":"Baylor College of Medicine","LAST_NAME":"Song","FIRST_NAME":"Shiyang","ADDRESS":"One Baylor Plaza, Houston, TX ,77030.","EMAIL":"shiyangs@bcm.edu","PHONE":"7137983159"},

"STUDY":{"STUDY_TITLE":"Myocardial Rev-erb-mediated diurnal metabolic rhythm (Part 3/3)","STUDY_SUMMARY":"The snap-frozen mouse heart ventricles that was harvested at ZT6 or ZT18 from both WT and Rev-erb cardiomyocytes specific KO mice were used for metabolomics study.","INSTITUTE":"Baylor College of Medicine","LAST_NAME":"Song","FIRST_NAME":"Shiyang","ADDRESS":"One Baylor Plaza","EMAIL":"shiyangs@bcm.edu","PHONE":"7137983159"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","AGE_OR_AGE_RANGE":"10 weeks","GENDER":"Male"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"WT_ZT6_1",
"Factors":{"Group":"WT_ZT6","Genotype":"wild type control","Experimental variables":"WT mouse heart ventrcle harvest at ZT6"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"WT_ZT6_2",
"Factors":{"Group":"WT_ZT6","Genotype":"wild type control","Experimental variables":"WT mouse heart ventrcle harvest at ZT6"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"WT_ZT6_3",
"Factors":{"Group":"WT_ZT6","Genotype":"wild type control","Experimental variables":"WT mouse heart ventrcle harvest at ZT6"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"KO_ZT6_1",
"Factors":{"Group":"KO_ZT6","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"KO mouse heart ventrcle harvest at ZT6"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"KO_ZT6_2",
"Factors":{"Group":"KO_ZT6","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"KO mouse heart ventrcle harvest at ZT6"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"KO_ZT6_3",
"Factors":{"Group":"KO_ZT6","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"KO mouse heart ventrcle harvest at ZT6"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"WT_ZT18_1",
"Factors":{"Group":"WT_ZT18","Genotype":"wild type control","Experimental variables":"WT mouse heart ventrcle harvest at ZT18"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"WT_ZT18_2",
"Factors":{"Group":"WT_ZT18","Genotype":"wild type control","Experimental variables":"WT mouse heart ventrcle harvest at ZT18"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"WT_ZT18_3",
"Factors":{"Group":"WT_ZT18","Genotype":"wild type control","Experimental variables":"WT mouse heart ventrcle harvest at ZT18"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"KO_ZT18_1",
"Factors":{"Group":"KO_ZT18","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"KO mouse heart ventrcle harvest at ZT18"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"KO_ZT18_2",
"Factors":{"Group":"KO_ZT18","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"KO mouse heart ventrcle harvest at ZT18"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
},
{
"Subject ID":"-",
"Sample ID":"KO_ZT18_3",
"Factors":{"Group":"KO_ZT18","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"KO mouse heart ventrcle harvest at ZT18"},
"Additional sample data":{"age(weeks)":"10","species":"mouse","strain":"C57bl/6","RAW_FILE_NAME":"RevCKOzt.zip","Tissue":"heart"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Snap-frozen heart ventricles were collected from both WT and Rev-erb cardiomyocyte-specific KO mice at ZT6 or ZT18","SAMPLE_TYPE":"Heart"},

"TREATMENT":{"TREATMENT_SUMMARY":"No special treatment"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Heart ventricle tissues were harvested from male mice at 2.5 months of age (n = 3 at each condition) and snap-frozen in liquid nitrogen. Data were acquired in multiple reaction monitoring (MRM) using Agilent QQQ LC-MS systems. Separation of TCA and glycolysis metabolites were performed using 5 mM ammonium acetate in water as buffer pH 9.9 (A), and 100% acetonitrile as buffer (B) using Luna 3 µM NH2 column (Phenomenex, Torrance, CA) and measured 6495 triple quadrupole mass spectrometer via ESI negative mode (Agilent Technologies, Santa Clara, CA). The Gradient used is as follows: 0-20 min-80% B (Flow rate 0.2ml/min); 20-20.10 min- 80% to 2 % B; 20.10-25 min-2% B (Flow rate 0.3ml/min); 25-30 min 80% B (Flowrate 0.35ml/min); 30-35 min-80%B (Flow rate 0.4ml/min); 35-38 min 80% B (Flow rate 0.4ml/min); followed by re-equilibration at the end of the gradient to the initial starting condition 80% B at Flow rate of 0.2 ml/min. Separation and measurement of amino acids were performed using Zorbax eclipse XDB C-18, 1.8 micron, 4.6 x 100 mm column on 6495 triple quadrupole mass spectrometer via ESI Positive mode (Agilent Technologies, Santa Clara, CA). Mobile phases A and B were 0.1% formic acid in water and acetonitrile, respectively. The gradient used is as follows: 0 min-2% B; 6 min- 2% of B, 6.5 min-30 % B, 7 min- 90% of B, 12 min 95% of B, 13 min 2% of B followed by re-equilibration at the end of the gradient 20 min to the initial starting condition 2% of B. Flow rate: 0.2 ml/min. Separation and measurement of CoA's and carnitines were performed using Zorbax eclipse XDB C-18, 1.8 micron, 4.6 x 100 mm column on 6495 triple quadrupole mass spectrometer via ESI Positive mode (Agilent Technologies, Santa Clara, CA). Mobile phases A and B were 0.1% formic acid in water and acetonitrile, respectively. Gradient used is as follows: 0 min-2% B; 6 min- 2% of B, 6.5 min-30 % B, 7 min- 90% of B, 12 min 95% of B,13 min 2% of B followed by re-equilibration at end of the gradient 20 min to the initial starting condition 2% of B. Flow rate: 0.2 ml/min. Separation and measurement of nucleotides were performed using Zorbax eclipse XDB C-18, 1.8 micron, 4.6 x 100 mm column on 6495 triple quadrupole mass spectrometer via ESI Positive mode (Agilent Technologies, Santa Clara, CA). Mobile phases A and B were 0.1% formic acid in water and acetonitrile, respectively. The gradient used is as follows: 0-6 min 2% B, 6-6.50 min 30% B, 7-12 min 95% B, 12-13 min 2% B, and re-equilibration till the end of the gradient 20 min. Flow rate: 0.2 ml/min). The data were normalized with internal standards, and log2 transformed on a per-sample, per-method basis. Statistical analyses were performed with either ANOVA or t-test in R Studio (R Studio Inc., Boston, MA). Differential metabolites were identified by adjusting the p-values for multiple testing at an FDR (Benjamini Hochberg method) threshold of <0.25."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Separation of TCA and glycolysis metabolites were performed using 5 mM ammonium acetate in water as buffer pH 9.9 (A), and 100% acetonitrile as buffer (B) using Luna 3 µM NH2 column (Phenomenex, Torrance, CA) and measured 6495 triple quadrupole mass spectrometer via ESI negative mode (Agilent Technologies, Santa Clara, CA). The Gradient used is as follows: 0-20 min-80% B (Flow rate 0.2ml/min); 20-20.10 min- 80% to 2 % B; 20.10-25 min-2% B (Flow rate 0.3ml/min); 25-30 min 80% B (Flowrate 0.35ml/min); 30-35 min-80%B (Flow rate 0.4ml/min); 35-38 min 80% B (Flow rate 0.4ml/min); followed by re-equilibration at the end of the gradient to the initial starting condition 80% B at Flow rate of 0.2 ml/min. Separation and measurement of amino acids were performed using Zorbax eclipse XDB C-18, 1.8 micron, 4.6 x 100 mm column on 6495 triple quadrupole mass spectrometer via ESI Positive mode (Agilent Technologies, Santa Clara, CA). Mobile phases A and B were 0.1% formic acid in water and acetonitrile, respectively. The gradient used is as follows: 0 min-2% B; 6 min- 2% of B, 6.5 min-30 % B, 7 min- 90% of B, 12 min 95% of B, 13 min 2% of B followed by re-equilibration at the end of the gradient 20 min to the initial starting condition 2% of B. Flow rate: 0.2 ml/min. Separation and measurement of CoA's and carnitines were performed using Zorbax eclipse XDB C-18, 1.8 micron, 4.6 x 100 mm column on 6495 triple quadrupole mass spectrometer via ESI Positive mode (Agilent Technologies, Santa Clara, CA). Mobile phases A and B were 0.1% formic acid in water and acetonitrile, respectively. Gradient used is as follows: 0 min-2% B; 6 min- 2% of B, 6.5 min-30 % B, 7 min- 90% of B, 12 min 95% of B,13 min 2% of B followed by re-equilibration at end of the gradient 20 min to the initial starting condition 2% of B. Flow rate: 0.2 ml/min. Separation and measurement of nucleotides were performed using Zorbax eclipse XDB C-18, 1.8 micron, 4.6 x 100 mm column on 6495 triple quadrupole mass spectrometer via ESI Positive mode (Agilent Technologies, Santa Clara, CA). Mobile phases A and B were 0.1% formic acid in water and acetonitrile, respectively. The gradient used is as follows: 0-6 min 2% B, 6-6.50 min 30% B, 7-12 min 95% B, 12-13 min 2% B, and re-equilibration till the end of the gradient 20 min. Flow rate: 0.2 ml/min).","CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Agilent 1290","COLUMN_NAME":"Agilent Zorbax Eclipse Plus C18 (100 x 2.1mm, 1.8 um)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Agilent 6490 QQQ","INSTRUMENT_TYPE":"Other","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"The data were normalized with internal standards, and log2 transformed on a per-sample, per-method basis. Statistical analyses were performed with either ANOVA or t-test in R Studio (R Studio Inc., Boston, MA). Differential metabolites were identified by adjusting the p-values for multiple testing at an FDR (Benjamini Hochberg method) threshold of <0.25."},

"MS_METABOLITE_DATA":{
"Units":"pmoles/l",

"Data":[{"Metabolite":"2-Methylbutyrylcarnitine","WT_ZT6_1":"-4.381892394","WT_ZT6_2":"-5.759223604","WT_ZT6_3":"-4.022115572","KO_ZT6_1":"-6.611316915","KO_ZT6_2":"-5.856014571","KO_ZT6_3":"-6.829029148","WT_ZT18_1":"-4.94537646","WT_ZT18_2":"-5.271659572","WT_ZT18_3":"-4.216510295","KO_ZT18_1":"-6.947553006","KO_ZT18_2":"-6.248109821","KO_ZT18_3":"-5.825351853"},{"Metabolite":"Acetyl Carnitine","WT_ZT6_1":"1.437288648","WT_ZT6_2":"1.161571474","WT_ZT6_3":"2.118072965","KO_ZT6_1":"-0.047179543","KO_ZT6_2":"0.443964199","KO_ZT6_3":"-0.123191204","WT_ZT18_1":"1.545488913","WT_ZT18_2":"1.563032667","WT_ZT18_3":"1.617972284","KO_ZT18_1":"0.568578265","KO_ZT18_2":"0.684880763","KO_ZT18_3":"0.648663001"},{"Metabolite":"Butyrylcarnitine","WT_ZT6_1":"0.79957427","WT_ZT6_2":"-1.206277582","WT_ZT6_3":"1.641288281","KO_ZT6_1":"-1.856052976","KO_ZT6_2":"-0.835876994","KO_ZT6_3":"-1.651374374","WT_ZT18_1":"0.823219112","WT_ZT18_2":"0.017610783","WT_ZT18_3":"1.419307705","KO_ZT18_1":"-2.456407746","KO_ZT18_2":"-0.996608052","KO_ZT18_3":"-0.290794307"},{"Metabolite":"Carnitine","WT_ZT6_1":"-0.454574609","WT_ZT6_2":"-0.120101098","WT_ZT6_3":"-0.550798232","KO_ZT6_1":"-0.978334992","KO_ZT6_2":"-1.076443217","KO_ZT6_3":"-1.021562248","WT_ZT18_1":"-0.766491629","WT_ZT18_2":"-0.745424274","WT_ZT18_3":"-0.823556575","KO_ZT18_1":"-1.005505851","KO_ZT18_2":"-1.428359307","KO_ZT18_3":"-1.474194894"},{"Metabolite":"Deoxy carnitine","WT_ZT6_1":"-4.270215246","WT_ZT6_2":"-4.630389035","WT_ZT6_3":"-4.165802406","KO_ZT6_1":"-4.534608539","KO_ZT6_2":"-4.400605024","KO_ZT6_3":"-4.557567078","WT_ZT18_1":"-4.842771791","WT_ZT18_2":"-4.692621761","WT_ZT18_3":"-4.735120937","KO_ZT18_1":"-4.504739664","KO_ZT18_2":"-4.795163334","KO_ZT18_3":"-4.860662234"},{"Metabolite":"Heptanoyl Carnitine","WT_ZT6_1":"-1.953976894","WT_ZT6_2":"-3.360012462","WT_ZT6_3":"-0.293653407","KO_ZT6_1":"-4.452936515","KO_ZT6_2":"-3.433761156","KO_ZT6_3":"-4.362551227","WT_ZT18_1":"-1.979745895","WT_ZT18_2":"-2.764643023","WT_ZT18_3":"-1.390416743","KO_ZT18_1":"-5.505356817","KO_ZT18_2":"-3.425929397","KO_ZT18_3":"-2.941035228"},{"Metabolite":"Isobutyryl Carnitine","WT_ZT6_1":"0.745098295","WT_ZT6_2":"-1.168470043","WT_ZT6_3":"1.558966339","KO_ZT6_1":"-1.810833461","KO_ZT6_2":"-0.745065301","KO_ZT6_3":"-1.601386126","WT_ZT18_1":"0.833923961","WT_ZT18_2":"0.163807374","WT_ZT18_3":"1.328746026","KO_ZT18_1":"-2.572366927","KO_ZT18_2":"-1.126784446","KO_ZT18_3":"-0.310307159"},{"Metabolite":"Isovaleryl Carnitine","WT_ZT6_1":"-1.826136317","WT_ZT6_2":"-3.176470708","WT_ZT6_3":"-1.473851848","KO_ZT6_1":"-3.980833289","KO_ZT6_2":"-3.28228625","KO_ZT6_3":"-4.245672173","WT_ZT18_1":"-2.254717815","WT_ZT18_2":"-2.521520933","WT_ZT18_3":"-1.634725019","KO_ZT18_1":"-4.267122196","KO_ZT18_2":"-3.648181991","KO_ZT18_3":"-3.264285456"},{"Metabolite":"Octanoylcarnitine","WT_ZT6_1":"-1.765036165","WT_ZT6_2":"-1.793690624","WT_ZT6_3":"-0.637901969","KO_ZT6_1":"-4.246861757","KO_ZT6_2":"-2.974874926","KO_ZT6_3":"-3.53222784","WT_ZT18_1":"-3.246067217","WT_ZT18_2":"-3.802741916","WT_ZT18_3":"-2.699957943","KO_ZT18_1":"-4.707918892","KO_ZT18_2":"-3.165178037","KO_ZT18_3":"-3.318248846"},{"Metabolite":"Palmitoylcarnitine","WT_ZT6_1":"-6.936214608","WT_ZT6_2":"-8.034754028","WT_ZT6_3":"-6.132519176","KO_ZT6_1":"-10.16476942","KO_ZT6_2":"-8.46623654","KO_ZT6_3":"-8.647707048","WT_ZT18_1":"-12.28908897","WT_ZT18_2":"-12.20072949","WT_ZT18_3":"-12.20821005","KO_ZT18_1":"-11.87945058","KO_ZT18_2":"-13.41919137","KO_ZT18_3":"-12.55290294"},{"Metabolite":"Propionyl 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}

}