{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002093","ANALYSIS_ID":"AN003419","VERSION":"1","CREATED_ON":"March 2, 2022, 7:16 am"},

"PROJECT":{"PROJECT_TITLE":"Lipidomics of High Fat vs Control Mice","PROJECT_TYPE":"Untargeted LC-MS/MS Lipidomics","PROJECT_SUMMARY":"Obesity exacerbates inflammation upon lung injury; however, the mechanisms by which obesity primes pulmonary dysregulation prior to injury are not well studied. Notably, little is known about how obesity dysregulates pulmonary polyunsaturated fatty acid (PUFA) metabolism that is central to inflammation initiation and resolution. Herein, we first show with mass spectrometry that a high fat diet (HFD) administered to C57BL/6J mice increases the relative abundance of pulmonary PUFA-containing triglycerides and the concentration of PUFA-derived oxylipins, independent of an increase in total pulmonary PUFAs. Experiments with a genetic model of obesity did not recapitulate the effects of the HFD on the pulmonary oxylipin signature, suggesting a diet-driven effect. Subsequent pulmonary next-generation RNA sequencing showed complex and unique transcriptional regulation with a HFD. The HFD increased pathways related to glycerophospholipid metabolism, innate immunity, and inflammation including an elevation in B cell differentiation and signaling. Finally, computational integration of lipidomic with transcriptomic data revealed novel networks with the HFD between glycerophospholipid metabolism and B cell receptor signaling with specific oxylipins. Collectively, these data show obesity dysregulates pulmonary PUFA metabolism prior to lung injury, which may be a mechanism by which obesity primes the lungs to respond poorly upon infectious and/or inflammatory challenges.","INSTITUTE":"University of North Carolina at Chapel Hill","DEPARTMENT":"Chemistry","LABORATORY":"MS Core Laboratory","LAST_NAME":"Weatherspoon","FIRST_NAME":"Emily","ADDRESS":"131 South Rd","EMAIL":"emdiane@email.unc.edu","PHONE":"7042453664"},

"STUDY":{"STUDY_TITLE":"Lipidomics of High Fat vs Control Mice","STUDY_TYPE":"Untargeted Lipidomics","STUDY_SUMMARY":"Obesity exacerbates inflammation upon lung injury; however, the mechanisms by which obesity primes pulmonary dysregulation prior to injury are not well studied. Notably, little is known about how obesity dysregulates pulmonary polyunsaturated fatty acid (PUFA) metabolism that is central to inflammation initiation and resolution. Herein, we first show with mass spectrometry that a high fat diet (HFD) administered to C57BL/6J mice increases the relative abundance of pulmonary PUFA-containing triglycerides and the concentration of PUFA-derived oxylipins, independent of an increase in total pulmonary PUFAs. Experiments with a genetic model of obesity did not recapitulate the effects of the HFD on the pulmonary oxylipin signature, suggesting a diet-driven effect. Subsequent pulmonary next-generation RNA sequencing showed complex and unique transcriptional regulation with a HFD. The HFD increased pathways related to glycerophospholipid metabolism, innate immunity, and inflammation including an elevation in B cell differentiation and signaling. Finally, computational integration of lipidomic with transcriptomic data revealed novel networks with the HFD between glycerophospholipid metabolism and B cell receptor signaling with specific oxylipins. Collectively, these data show obesity dysregulates pulmonary PUFA metabolism prior to lung injury, which may be a mechanism by which obesity primes the lungs to respond poorly upon infectious and/or inflammatory challenges.","INSTITUTE":"University of North Carolina at Chapel Hill","DEPARTMENT":"Chemistry","LABORATORY":"MS Core Laboratory","LAST_NAME":"Weatherspoon","FIRST_NAME":"Emily","ADDRESS":"131 South Rd","EMAIL":"emdiane@email.unc.edu","PHONE":"7042453664","NUM_GROUPS":"2","TOTAL_SUBJECTS":"3","NUM_MALES":"6"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","AGE_OR_AGE_RANGE":"21 weeks","GENDER":"Male"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"HF-R-4R",
"Factors":{"Treatment":"High Fat"},
"Additional sample data":{"RAW_FILE_NAME":"HF-R-4R.raw"}
},
{
"Subject ID":"-",
"Sample ID":"HF-R-4L",
"Factors":{"Treatment":"High Fat"},
"Additional sample data":{"RAW_FILE_NAME":"HF-R-4L.raw"}
},
{
"Subject ID":"-",
"Sample ID":"HF-L-5L",
"Factors":{"Treatment":"High Fat"},
"Additional sample data":{"RAW_FILE_NAME":"HF-L-5L.raw"}
},
{
"Subject ID":"-",
"Sample ID":"HF-L-4RL",
"Factors":{"Treatment":"High Fat"},
"Additional sample data":{"RAW_FILE_NAME":"HF-L-4RL.raw"}
},
{
"Subject ID":"-",
"Sample ID":"HF-L-4R",
"Factors":{"Treatment":"High Fat"},
"Additional sample data":{"RAW_FILE_NAME":"HF-L-4R.raw"}
},
{
"Subject ID":"-",
"Sample ID":"HF-L-4L",
"Factors":{"Treatment":"High Fat"},
"Additional sample data":{"RAW_FILE_NAME":"HF-L-4L.raw"}
},
{
"Subject ID":"-",
"Sample ID":"HF-L-3MN",
"Factors":{"Treatment":"High Fat"},
"Additional sample data":{"RAW_FILE_NAME":"HF-L-3MN.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CON-R-8RL",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"CON-R-8RL.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CON-R-7RN",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"CON-R-7RN.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CON-L-8RL",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"CON-L-8RL.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CON-L-8LN",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"CON-L-8LN.raw"}
},
{
"Subject ID":"-",
"Sample ID":"Con-L-7RN",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"con-L-7RN.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CON-L-7NW",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"CON-L-7NW.raw"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"C57BL/6J male mice were purchased from Jackson Laboratories. At 6 weeks of age, mice were fed lean control (10% kcal from fat) or high-fat diet (60% kcal from fat) for 15 weeks (Research Diets, New Brunswick, NJ). Ob/ob mice, a genetic model of obesity driven by leptin deficiency, were purchased from Jackson laboratories at age 10 weeks and were fed a normal chow diet for an additional 5 weeks.","SAMPLE_TYPE":"Lung"},

"TREATMENT":{"TREATMENT_SUMMARY":"C57BL/6J male mice were purchased from Jackson Laboratories. At 6 weeks of age, mice were fed lean control (10% kcal from fat) or high-fat diet (60% kcal from fat) for 15 weeks (Research Diets, New Brunswick, NJ). Ob/ob mice, a genetic model of obesity driven by leptin deficiency, were purchased from Jackson laboratories at age 10 weeks and were fed a normal chow diet for an additional 5 weeks."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Mouse lungs were weighed into Eppendorf tubes. The lung tissues were mashed using a clean metal spatula prior to extraction. The samples were extracted using a liquid-liquid partition with water (250 µL), methanol (300 µL), and methyl tert-butyl ether (MTBE). Avanti’s deuterated lipid mix, Equisplash, was used as an internal standard. The deuterated mix was spiked into the methanol at 1.5 µg/mL and used for extraction. The extracts were centrifuged at 20,000 rcf for 10 minutes to facilitate phase separation. The top layer was removed, dried, and reconstituted in 150 µL of isopropanol for analysis.","PROCESSING_STORAGE_CONDITIONS":"-80℃","SAMPLE_RESUSPENSION":"150 µL of IPA"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Waters Acquity I-Class","COLUMN_NAME":"Waters Acquity BEH C18 (100 x 2mm, 1.7um)","FLOW_GRADIENT":"Starting composition was 32% B, which increased to 40% B at 1 min (held until 1.5 min) then 45% B at 4 minutes. This was increased to 50% B at 5 min, 60% B at 8 min, 70% B at 11 min, and 80% B at 14 min (held until 16 min). At 16 min the composition switched back to starting conditions (32% B) and was held for 4 min to re-equilibrate the column.","FLOW_RATE":"0.2 mL/min","COLUMN_TEMPERATURE":"45","SOLVENT_A":"60/40 ACN/H20 10 mM ammonium formate","SOLVENT_B":"B- 90/10 IPA/ACN 10 mM ammonium formate","INJECTION_TEMPERATURE":"10"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","DATA_FORMAT":".Raw"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive HF-X Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Samples were analyzed with positive/negative ionization switching. 60,000 resolution. Top 5 DDA.","COLLISION_ENERGY":"25, 35, 45 V stepped collision energy","FRAGMENTATION_METHOD":"HCD","IONIZATION":"HESI","MASS_ACCURACY":"5 ppm"},

"MS_METABOLITE_DATA":{
"Units":"Peak Area",

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}

}