{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001916","ANALYSIS_ID":"AN003114","VERSION":"1","CREATED_ON":"September 13, 2021, 5:28 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","EMAIL":"shiyangs@bcm.edu","PHONE":"7137983159"},

"STUDY":{"STUDY_TITLE":"Myocardial Rev-erb-mediated diurnal metabolic rhythm (Part 2/3)","STUDY_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"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"WT_CD_1",
"Factors":{"Group":"WT_CD","Genotype":"wild type control","Experimental variables":"heart ventrcle harvest from WT mouse  fed with chow diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"WT_CD_2",
"Factors":{"Group":"WT_CD","Genotype":"wild type control","Experimental variables":"heart ventrcle harvest from WT mouse  fed with chow diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"WT_CD_3",
"Factors":{"Group":"WT_CD","Genotype":"wild type control","Experimental variables":"heart ventrcle harvest from WT mouse  fed with chow diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"KO_CD_1",
"Factors":{"Group":"KO_CD","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"heart ventrcle harvest from KO mouse  fed with chow diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"KO_CD_2",
"Factors":{"Group":"KO_CD","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"heart ventrcle harvest from KO mouse  fed with chow diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"KO_CD_3",
"Factors":{"Group":"KO_CD","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"heart ventrcle harvest from KO mouse  fed with chow diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"WT_HFHSD_1",
"Factors":{"Group":"WT_HFHSD","Genotype":"wild type control","Experimental variables":"heart ventrcle harvest from WT mouse  fed with high fat high sucrose diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"WT_HFHSD_2",
"Factors":{"Group":"WT_HFHSD","Genotype":"wild type control","Experimental variables":"heart ventrcle harvest from WT mouse  fed with high fat high sucrose diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"WT_HFHSD_3",
"Factors":{"Group":"WT_HFHSD","Genotype":"wild type control","Experimental variables":"heart ventrcle harvest from WT mouse  fed with high fat high sucrose diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"KO-HFHSD_1",
"Factors":{"Group":"KO-HFHSD","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"heart ventrcle harvest from KOmouse  fed with high fat high sucrose diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"KO-HFHSD_2",
"Factors":{"Group":"KO-HFHSD","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"heart ventrcle harvest from KOmouse  fed with high fat high sucrose diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
},
{
"Subject ID":"-",
"Sample ID":"KO-HFHSD_3",
"Factors":{"Group":"KO-HFHSD","Genotype":"Rev-erb cardiomyocyte specific knock out","Experimental variables":"heart ventrcle harvest from KOmouse  fed with high fat high sucrose diet"},
"Additional sample data":{"age(months)":"4.5","species":"mouse","strain":"C57bl/6","tissue":"heart","RAW_FILE_NAME":"RevCKOdiet.zip"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Snap-frozen heart ventricles were collected from both WT and Rev-erb cardiomyocyte-specific KO mice fed with either chow diet or high fat high sucrose diet(HFHSD), HFHSD start at 8 weeks old and mice were harvest at 4.5 months of age.","SAMPLE_TYPE":"Heart"},

"TREATMENT":{"TREATMENT_SUMMARY":"mice were fed with either the chow diet or high-fat high sucrose diet as indicated."},

"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":"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_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":"Alanine","WT_CD_1":"-2.220278098","WT_CD_2":"-1.64067611","WT_CD_3":"-1.284423874","KO_CD_1":"-1.701284354","KO_CD_2":"-1.987770791","KO_CD_3":"-1.686866086","WT_HFHSD_1":"-2.177782496","WT_HFHSD_2":"-1.932152075","WT_HFHSD_3":"-1.661779595","KO-HFHSD_1":"-1.695203748","KO-HFHSD_2":"-1.802323601","KO-HFHSD_3":"-2.297759189"},{"Metabolite":"Arginine","WT_CD_1":"-1.92296464","WT_CD_2":"-1.555360589","WT_CD_3":"-1.434931304","KO_CD_1":"-2.557416109","KO_CD_2":"-2.128414108","KO_CD_3":"-1.791413965","WT_HFHSD_1":"-1.400997564","WT_HFHSD_2":"-1.472726624","WT_HFHSD_3":"-1.013927481","KO-HFHSD_1":"-1.216499826","KO-HFHSD_2":"-1.432509191","KO-HFHSD_3":"-1.28412705"},{"Metabolite":"Asparagine","WT_CD_1":"-0.159295876","WT_CD_2":"-0.65904227","WT_CD_3":"-0.503348945","KO_CD_1":"-0.328640024","KO_CD_2":"-0.45440137","KO_CD_3":"0.075477381","WT_HFHSD_1":"0.22649214","WT_HFHSD_2":"-0.575614905","WT_HFHSD_3":"-0.174352573","KO-HFHSD_1":"-0.045002976","KO-HFHSD_2":"-0.935162288","KO-HFHSD_3":"-1.372765701"},{"Metabolite":"Aspartic acid","WT_CD_1":"-3.7233828","WT_CD_2":"-4.086707945","WT_CD_3":"-4.024967702","KO_CD_1":"-3.110121437","KO_CD_2":"-3.016956848","KO_CD_3":"-2.43531228","WT_HFHSD_1":"-4.704395227","WT_HFHSD_2":"-4.557583164","WT_HFHSD_3":"-4.482096695","KO-HFHSD_1":"-3.495198668","KO-HFHSD_2":"-3.631666293","KO-HFHSD_3":"-3.798707691"},{"Metabolite":"Betaine","WT_CD_1":"0.206761827","WT_CD_2":"-0.290343211","WT_CD_3":"-1.005269562","KO_CD_1":"0.207507327","KO_CD_2":"0.363120175","KO_CD_3":"0.164161965","WT_HFHSD_1":"-1.050198518","WT_HFHSD_2":"-1.58412704","WT_HFHSD_3":"-1.011811928","KO-HFHSD_1":"0.256616199","KO-HFHSD_2":"0.117070313","KO-HFHSD_3":"-0.130884639"},{"Metabolite":"Citrulline","WT_CD_1":"-4.463614704","WT_CD_2":"-4.264140452","WT_CD_3":"-3.796612194","KO_CD_1":"-3.850167223","KO_CD_2":"-3.765672174","KO_CD_3":"-3.402661076","WT_HFHSD_1":"-2.707146545","WT_HFHSD_2":"-3.140183397","WT_HFHSD_3":"-2.872855341","KO-HFHSD_1":"-2.570382924","KO-HFHSD_2":"-2.769973901","KO-HFHSD_3":"-2.591729463"},{"Metabolite":"Cystine","WT_CD_1":"-6.838751662","WT_CD_2":"-7.209769871","WT_CD_3":"-6.888871463","KO_CD_1":"-7.769905585","KO_CD_2":"-7.22343719","KO_CD_3":"-7.104681619","WT_HFHSD_1":"-6.914813848","WT_HFHSD_2":"-7.975952217","WT_HFHSD_3":"-7.184332423","KO-HFHSD_1":"-7.290334575","KO-HFHSD_2":"-7.54466992","KO-HFHSD_3":"-7.656977219"},{"Metabolite":"Glucosamine","WT_CD_1":"-6.340338736","WT_CD_2":"-6.250981917","WT_CD_3":"-6.385005879","KO_CD_1":"-6.389096605","KO_CD_2":"-6.564978313","KO_CD_3":"-6.238704219","WT_HFHSD_1":"-6.26312481","WT_HFHSD_2":"-6.266873592","WT_HFHSD_3":"-6.33204374","KO-HFHSD_1":"-6.049868081","KO-HFHSD_2":"-6.624333307","KO-HFHSD_3":"-6.268866803"},{"Metabolite":"Glutaric 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"Metabolites":[{"metabolite_name":"Alanine"},{"metabolite_name":"Arginine"},{"metabolite_name":"Asparagine"},{"metabolite_name":"Aspartic acid"},{"metabolite_name":"Betaine"},{"metabolite_name":"Citrulline"},{"metabolite_name":"Cystine"},{"metabolite_name":"Glucosamine"},{"metabolite_name":"Glutaric acid"},{"metabolite_name":"Glycine"},{"metabolite_name":"Guanine"},{"metabolite_name":"Histidine"},{"metabolite_name":"Homocysteine"},{"metabolite_name":"Isoleucine"},{"metabolite_name":"Lysine"},{"metabolite_name":"Methionine"},{"metabolite_name":"N-acetyl AA"},{"metabolite_name":"Ornithine"},{"metabolite_name":"Phenyl alanine"},{"metabolite_name":"Proline"},{"metabolite_name":"Putrescine"},{"metabolite_name":"Serine"},{"metabolite_name":"Spermidine"},{"metabolite_name":"Spermine"},{"metabolite_name":"Threonine"},{"metabolite_name":"Tryptophan"},{"metabolite_name":"Tyrosine"},{"metabolite_name":"Valine"},{"metabolite_name":"2-Methylbutyrylcarnitine"},{"metabolite_name":"Acetyl Carnitine"},{"metabolite_name":"Butyrylcarnitine"},{"metabolite_name":"Deoxy carnitine"},{"metabolite_name":"GlutaroylCoA"},{"metabolite_name":"Carnitine"},{"metabolite_name":"Heptanoyl Carnitine"},{"metabolite_name":"Isobutyryl Carnitine"},{"metabolite_name":"Isovaleryl Carnitine"},{"metabolite_name":"Malonylcarnitine"},{"metabolite_name":"Octanoylcarnitine"},{"metabolite_name":"MethylGlutaryl carnitine"},{"metabolite_name":"Palmitoylcarnitine"},{"metabolite_name":"Palmityl Carnitine"},{"metabolite_name":"Propionyl Carnitine"},{"metabolite_name":"Succinyl CoA"},{"metabolite_name":"3PG and 2PG"},{"metabolite_name":"Citrate"},{"metabolite_name":"Erythrose-4-P"},{"metabolite_name":"FBP/GBP"},{"metabolite_name":"Fumarate"},{"metabolite_name":"G6P/F6P"},{"metabolite_name":"GlucoseFructose"},{"metabolite_name":"Glutamic acid"},{"metabolite_name":"Glutamine"},{"metabolite_name":"Glycerol 3 Phosphate"},{"metabolite_name":"Hydroxy glutarate"},{"metabolite_name":"Ketoglutarate"},{"metabolite_name":"Lactate"},{"metabolite_name":"Malate"},{"metabolite_name":"OAA"},{"metabolite_name":"Ribose/Ribulose/xylulose-5P"},{"metabolite_name":"Sedoheptulose-7-P"},{"metabolite_name":"Succinate"}]
}

}