{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002303","ANALYSIS_ID":"AN003764","VERSION":"1","CREATED_ON":"October 6, 2022, 9:40 am"},

"PROJECT":{"PROJECT_TITLE":"Fitm2 is required for ER homeostasis and normal function of murine liver","PROJECT_SUMMARY":"The ER-resident protein fat-inducing transcript 2 (FIT2) catalyzes acyl-CoA cleavage in vitro and is required for endoplasmic reticulum (ER) homeostasis and normal lipid storage in cells. The gene encoding FIT2 is essential for the viability of mice and worms. Whether FIT2 acts as an acyl-CoA diphosphatase in vivo and how this activity affects liver, where the protein was discovered, are unknown. Here, we report that hepatocyte-specific Fitm2 knockout (FIT2-LKO) mice fed a chow diet exhibited elevated acyl-CoA levels, ER stress, and signs of liver injury. These mice also had more triglycerides in their livers than control littermates due, in part, to impaired secretion of triglyceride-rich lipoproteins and reduced capacity for fatty acid oxidation. Challenging FIT2-LKO mice with a high-fat diet worsened hepatic ER stress and liver injury, but unexpectedly reversed the steatosis phenotype, similar to what is observed in FIT2-deficient cells loaded with fatty acids. Our findings support the model that FIT2 acts as an acyl-CoA diphosphatase in vivo and is crucial for normal hepatocyte function and ER homeostasis in murine liver.","INSTITUTE":"Harvard University","DEPARTMENT":"Molecular Metabolism","LABORATORY":"FareseWalther's lab","LAST_NAME":"Bond","FIRST_NAME":"Laura","ADDRESS":"655 Huntington Avenue, Building 1, Room 203, Boston, MA, 02115, USA","EMAIL":"laurabond44@gmail.com","PHONE":"+18573087183"},

"STUDY":{"STUDY_TITLE":"Fitm2 is required for ER homeostasis and normal function of murine liver","STUDY_SUMMARY":"The ER-resident protein fat-inducing transcript 2 (FIT2) catalyzes acyl-CoA cleavage in vitro and is required for endoplasmic reticulum (ER) homeostasis and normal lipid storage in cells. The gene encoding FIT2 is essential for the viability of mice and worms. Whether FIT2 acts as an acyl-CoA diphosphatase in vivo and how this activity affects liver, where the protein was discovered, are unknown. Here, we report that hepatocyte-specific Fitm2 knockout (FIT2-LKO) mice fed a chow diet exhibited elevated acyl-CoA levels, ER stress, and signs of liver injury. These mice also had more triglycerides in their livers than control littermates due, in part, to impaired secretion of triglyceride-rich lipoproteins and reduced capacity for fatty acid oxidation. Challenging FIT2-LKO mice with a high-fat diet worsened hepatic ER stress and liver injury, but unexpectedly reversed the steatosis phenotype, similar to what is observed in FIT2-deficient cells loaded with fatty acids. Our findings support the model that FIT2 acts as an acyl-CoA diphosphatase in vivo and is crucial for normal hepatocyte function and ER homeostasis in murine liver.","INSTITUTE":"Harvard School of Public Health","LAST_NAME":"Bond","FIRST_NAME":"Laura","ADDRESS":"665 Huntington Ave., Building 2, 3rd Floor, Room 311 | Boston, MA 02115","EMAIL":"laurabond44@gmail.com","PHONE":"8573087183"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Blank",
"Factors":{"Genotype":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"blank"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2Flox_01",
"Factors":{"Genotype":"flox"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2Flox_01"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2Flox_02",
"Factors":{"Genotype":"flox"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2Flox_02"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2Flox_03",
"Factors":{"Genotype":"flox"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2Flox_03"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2Flox_04",
"Factors":{"Genotype":"flox"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2Flox_04"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2Flox_05",
"Factors":{"Genotype":"flox"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2Flox_05"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2-LKO_01",
"Factors":{"Genotype":"Liver Knockout"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2-LKO_01"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2-LKO_02",
"Factors":{"Genotype":"Liver Knockout"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2-LKO_02"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2-LKO_03",
"Factors":{"Genotype":"Liver Knockout"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2-LKO_03"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2-LKO_04",
"Factors":{"Genotype":"Liver Knockout"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2-LKO_04"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2-LKO_05",
"Factors":{"Genotype":"Liver Knockout"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2-LKO_05"}
},
{
"Subject ID":"-",
"Sample ID":"FIT2-LKO_06",
"Factors":{"Genotype":"Liver Knockout"},
"Additional sample data":{"RAW_FILE_NAME":"FIT2-LKO_06"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"All mice fasted 2 hours prior to sacrifice. Mice were euthanized with isoflurane and tissues were collected.","SAMPLE_TYPE":"Liver"},

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

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Liver (~100mg) was homogenized in 1 mL of PBS using a Bead Mill Homogenizer (VWR). Lipids were extracted, according to the Folch method.","PROCESSING_STORAGE_CONDITIONS":"On ice","EXTRACT_STORAGE":"-80℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Thermo Accucore C30, 2.1 x 250 mm, 3 μm"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"Mass spectrometry data analysis was performed using LipidSearch version 4.1 SP (Thermo Fisher Scientific). The results were exported to R-Studio where quality control was performed using pairwise correlations between replicates, a principal component analysis comparing sample groups, as well as retention time plot analysis to verify elution clustering within lipid classes. All identified lipids were included for subsequent analyses if they fulfilled the following LipidSearch-based criteria: 1) reject equal to zero, 2) main grade A OR main grade B AND APvalue<0.01 for at least three replicates, and 3) no missing values across all samples."},

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

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":"PI(18:0/18:1)"},{"metabolite_name":"PI(18:0/18:2)"},{"metabolite_name":"PI(18:0/20:2)"},{"metabolite_name":"PI(18:0/20:3)"},{"metabolite_name":"PI(18:0/20:4)"},{"metabolite_name":"PI(18:0/20:5)"},{"metabolite_name":"PI(18:0/22:4)"},{"metabolite_name":"PI(18:0/22:5)"},{"metabolite_name":"PI(18:0/22:6)"},{"metabolite_name":"PI(18:0p/20:4)"},{"metabolite_name":"PI(18:1/18:1)"},{"metabolite_name":"PI(18:1/18:2)"},{"metabolite_name":"PI(18:1/20:3)"},{"metabolite_name":"PI(18:1/20:4)"},{"metabolite_name":"PI(18:1/22:5)"},{"metabolite_name":"PI(18:1/22:6)"},{"metabolite_name":"PI(18:2/18:2)"},{"metabolite_name":"PI(18:2/20:4)"},{"metabolite_name":"PI(18:3/18:2)"},{"metabolite_name":"PI(18:3/20:4)"},{"metabolite_name":"PI(19:0/20:3)"},{"metabolite_name":"PI(19:0/20:4)"},{"metabolite_name":"PI(19:1/20:4)"},{"metabolite_name":"PI(20:0/20:3)"},{"metabolite_name":"PI(20:0/20:4)"},{"metabolite_name":"PI(20:4/20:4)"},{"metabolite_name":"PS(16:0/18:2)"},{"metabolite_name":"PS(16:0/20:3)"},{"metabolite_name":"PS(16:0/20:4)"},{"metabolite_name":"PS(16:0/20:5)"},{"metabolite_name":"PS(16:0/22:6)"},{"metabolite_name":"PS(17:0/22:6)"},{"metabolite_name":"PS(18:0/18:1)"},{"metabolite_name":"PS(18:0/18:2)"},{"metabolite_name":"PS(18:0/20:3)"},{"metabolite_name":"PS(18:0/20:4)"},{"metabolite_name":"PS(18:0/20:5)"},{"metabolite_name":"PS(18:0/22:4)"},{"metabolite_name":"PS(18:0/22:5)"},{"metabolite_name":"PS(18:0/22:6)"},{"metabolite_name":"PS(18:1/22:6)"},{"metabolite_name":"PS(18:2/18:2)"},{"metabolite_name":"PS(19:0/22:6)"},{"metabolite_name":"PS(20:0/22:6)"},{"metabolite_name":"PS(20:2/20:2)"},{"metabolite_name":"PS(20:4/22:6)"},{"metabolite_name":"PS(20:5/20:4)"},{"metabolite_name":"SM(d17:1/16:0)"},{"metabolite_name":"SM(d18:1/16:0)"},{"metabolite_name":"SM(d18:1/24:1)"},{"metabolite_name":"SM(d18:1/24:3)"},{"metabolite_name":"SM(d18:1/26:4)"},{"metabolite_name":"SM(d20:0/18:1)"},{"metabolite_name":"SM(d20:0+pO/20:0)"},{"metabolite_name":"SM(d22:0/18:2)"},{"metabolite_name":"SM(d22:1/18:1)"},{"metabolite_name":"SM(d22:1/20:1)"},{"metabolite_name":"SM(d22:1/20:2)"}]
}

}