{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001061","ANALYSIS_ID":"AN001732","VERSION":"1","CREATED_ON":"September 21, 2018, 2:33 pm"},

"PROJECT":{"PROJECT_TITLE":"Multi-omics Approach Reveals Metabolic Changes in the Heart at Birth","PROJECT_SUMMARY":"During late gestation, the fetal heart primarily relies on glucose and lactate to support rapid growth and development. While numerous studies describe changes in heart metabolism a few weeks after birth to preferentially utilize fatty acids, little is known about metabolic changes of the heart within the first day following birth. Therefore, we used the ovine model of pregnancy to investigate metabolic differences between the near-term fetal and the newborn heart. Samples were collected for metabolomic, lipidomic, and transcriptomic approaches from the left and right ventricles and intraventricular septum in 7 fetuses at gestational day 142 and 7 newborn lambs on the day of birth. We observed greater abundance of metabolites involved in butanoate and propanoate metabolism, and glycolysis in the term fetal heart (FDR-corrected p<0.10) and differential expression in these pathways were confirmed with single-sample gene set enrichment analysis (ssGSEA) (FDR-corrected p<0.05). Immediately following birth, newborn hearts displayed enrichment in purine, fatty acid, and glycerophospholipid metabolic pathways, as well as oxidative phosphorylation with significant alterations in both lipids and metabolites to support transcriptomic findings. While other studies suggest a switch from carbohydrate metabolism to fatty acid metabolism in the neonatal heart in as early as 2 weeks following birth, our data show that this metabolic switch in the heart begins by the first day of postnatal life. A better understanding of metabolic alterations that occur in the heart following birth may improve treatment of neonates at risk for heart failure.","INSTITUTE":"University of Florida","DEPARTMENT":"Pharmacodynamics","LAST_NAME":"Keller-Wood","FIRST_NAME":"Maureen","ADDRESS":"1345 SW Archer Rd, PO 100487, Gainesville, FL, 32610","EMAIL":"kellerwd@cop.ufl.edu","PHONE":"NA"},

"STUDY":{"STUDY_TITLE":"Lipidomics of Near-Term Fetal and Newborn Sheep Cardiac Tissue","STUDY_TYPE":"Comparison","STUDY_SUMMARY":"Cardiac tissue from near-term fetal and newborn sheep were compared via NMR metabolomic analysis","INSTITUTE":"University of Florida","DEPARTMENT":"Biochemistry & Molecular Biology","LAST_NAME":"Walejko","FIRST_NAME":"Jacquelyn","ADDRESS":"R3-226 Academic Research Building, Department of Biochemistry and Molecular Biology, PO Box 100245, Gainesville, FL 32610-0245","EMAIL":"jwalejko@ufl.edu","PHONE":"na","NUM_GROUPS":"2","TOTAL_SUBJECTS":"14"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Ovis aries","TAXONOMY_ID":"9940","AGE_OR_AGE_RANGE":"142 days gestation (fetal); Within 24 hours of birth (newborn)"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"1355A",
"Sample ID":"1355A_RV",
"Factors":{"Age":"Fetal","Sex":"Male"}
},
{
"Subject ID":"1355B",
"Sample ID":"1355B_RV",
"Factors":{"Age":"Fetal","Sex":"Male"}
},
{
"Subject ID":"1555",
"Sample ID":"1555_RV",
"Factors":{"Age":"Fetal","Sex":"Male"}
},
{
"Subject ID":"1237A",
"Sample ID":"1237A_RV",
"Factors":{"Age":"Fetal","Sex":"Male"}
},
{
"Subject ID":"1562",
"Sample ID":"1562_RV",
"Factors":{"Age":"Fetal","Sex":"Male"}
},
{
"Subject ID":"1237B",
"Sample ID":"1237B_RV",
"Factors":{"Age":"Fetal","Sex":"Female"}
},
{
"Subject ID":"1477",
"Sample ID":"1477_RV",
"Factors":{"Age":"Newborn","Sex":"Male"}
},
{
"Subject ID":"1462",
"Sample ID":"1462_RV",
"Factors":{"Age":"Newborn","Sex":"Male"}
},
{
"Subject ID":"1481",
"Sample ID":"1481_RV",
"Factors":{"Age":"Fetal","Sex":"Female"}
},
{
"Subject ID":"5562A",
"Sample ID":"5562A_RV",
"Factors":{"Age":"Newborn","Sex":"Female"}
},
{
"Subject ID":"1767A",
"Sample ID":"1767A_RV",
"Factors":{"Age":"Newborn","Sex":"Female"}
},
{
"Subject ID":"5562C",
"Sample ID":"5562C_RV",
"Factors":{"Age":"Newborn","Sex":"Female"}
},
{
"Subject ID":"1767B",
"Sample ID":"1767B_RV",
"Factors":{"Age":"Newborn","Sex":"Female"}
},
{
"Subject ID":"5562B",
"Sample ID":"5562B_RV",
"Factors":{"Age":"Newborn","Sex":"Male"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Ewes were assigned to one of two groups: (1) a fetal group (n=7, 3 singleton and 2 multiple gestation pregnancies); (2) a newborn group (n=7, 2 singleton and 2 multiple gestation pregnancies). Heart tissue was collected from the right ventricle (RV), left ventricle (LV), and intraventricular septum (IVS) on the day of birth (newborn group) or at gestational day 142 (fetal group) and immediately frozen in liquid nitrogen. Samples were stored at -80 °C until data collection.","SAMPLE_TYPE":"Cardiac tissue","COLLECTION_METHOD":"Heart tissue was collected under sterile conditions and immediately frozen in liquid nitrogen","COLLECTION_LOCATION":"University of Florida","STORAGE_CONDITIONS":"-80℃","COLLECTION_VIALS":"Cryovials","STORAGE_VIALS":"Cryovials"},

"TREATMENT":{"TREATMENT_SUMMARY":"NA"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Homogenized right ventricle tissue samples (100 mg from 7 fetuses and 7 newborns) were used for lipid extraction via the Folch method. Briefly, 2 mL of internal standard mixture was added to each sample before adding 1.2 mL of 2:1 chloroform/methanol solution (HPLC-grade). Samples were incubated at 4 °C for 20 min with occasional vortexing. Following incubation, 200 mL of water (HPLC-grade) was added and samples were incubated at 4 °C for 10 min. Samples were centrifuged at 2,000 g for 5 min at 4 °C, and the resulting chloroform layer was removed and transferred into a clean tube. The extraction process was repeated with 400 mL 2:1 chloroform/methanol (HPLC-grade) and the resulting chloroform layers were combined. Samples were dried under a nitrogen stream at 30 °C and stored at -80 °C until reconstitution. Samples were reconstituted with 200 mL of isopropanol and 2 mL injection standard mixture. Ammonium acetate and all analytical grade solvents (formic acid, chloroform, and methanol) were purchased from Fisher Scientific (Waltham, MA, USA). All mobile phase solvents were Fisher Optima LC/MS-grade (acetonitrile, isopropanol, and water).","PROCESSING_METHOD":"50 mg of tissue was weighed and added to tubes with three 3mm glass beads, two 5mm steal beads, and 0.7mm zirconia beads (“2 squirts”). HPLC-water (1mL) was added to each tube before homogenization at 1800 rpm for 30 seconds for 5 rounds. Samples were incubated at 4C for 20 min between rounds. Following homogenization, samples were centrifuged at 2,000g for 10 min at 4C to pellet tissue debris before transferring supernatant to a clean tube. Homogenates were stored at -80C.","EXTRACTION_METHOD":"2 uL of internal standard mixture was added to each sample before adding 1.2 mL of 2:1 chloroform/methanol solution (HPLC-grade). Samples were incubated at 4 °C for 20 min with occasional vortexing. Following incubation, 200 mL of water (HPLC-grade) was added and samples were incubated at 4 °C for 10 min. Samples were centrifuged at 2,000 g for 5 min at 4 °C, and the resulting chloroform layer was removed and transferred into a clean tube. The extraction process was repeated with 400 mL 2:1 chloroform/methanol (HPLC-grade) and the resulting chloroform layers were combined. Samples were dried under a nitrogen stream at 30 °C (Organomation Associated MultiVap) and stored at -80 °C until reconstitution.","SAMPLE_RESUSPENSION":"Samples were reconstituted with 200 mL of isopropanol and 2 mL injection standard mixture and vortexed to mix. Fifty uL of reconstituted sample was transferred to a glass LC vial for analysis.","SAMPLE_SPIKING":"Internal standards: lysophosphatidylcholine (17:0), phosphatidylcholine (17:0/17:0), phosphatidylglycerol (14:0/14:0), phosphatidylethanolamine (15:0/15:0), phosphatidylserine (14:0/14:0), triglyceride (15:0/15:0/15:0). Injection Standards: lysophosphatidylcholine (19:0), phosphatidylcholine (19:0/19:0), phosphatidylglycerol (17:0/17:0), phosphatidylethanolamine (17:0/17:0), phosphatidylserine (17:0/17:0), triglyceride (17:0/17:0/17:0)."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Normal phase","INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","COLUMN_NAME":"Waters Acquity BEH C18 (50 x 2.1mm, 1.7um)","FLOW_GRADIENT":"80% A and 20% B at 0 min; 80% A and 20% B at 1 min; 70% A and 30% B at 3 min; 55% A and 45% B at 4 min; 40% A and 60% B at 6 min; 35% A and 65% B at 8 min; 35% A and 65% B at 10 min; 10% A and 90% B at 15 min; 2% A and 98% B at 17 min; 2% A and 98% B at 18 min; 80% A and 20% B at 19 min; 80% A and 20% B at 23 min","FLOW_RATE":"0.5mL/min","COLUMN_TEMPERATURE":"4C","SOLVENT_A":"Acetonitrile:H2O (60:40) with 10mM ammonium formate and 0.1% formic acid","SOLVENT_B":"Isopropanol:acetonitrile:H2O (90:8:2) with 10mM ammonium formate and 0.1% formic acid","SAMPLE_INJECTION":"2 uL","ANALYTICAL_TIME":"18 min"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"Timothy Garrett","OPERATOR_NAME":"Jeremy Koelmel"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","CAPILLARY_TEMPERATURE":"250C","IONIZATION":"Heated electrospray","SPRAY_VOLTAGE":"1500","RESOLUTION_SETTING":"70,000","SCAN_RANGE_MOVERZ":"200-2200 m/z"},

"MS_METABOLITE_DATA":{
"Units":"Relative peak area",

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