{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002314","ANALYSIS_ID":"AN003781","VERSION":"1","CREATED_ON":"October 13, 2022, 11:26 am"},

"PROJECT":{"PROJECT_TITLE":"Differential requirements for mitochondrial electron transport chain components in the adult murine liver","PROJECT_SUMMARY":"Mitochondrial electron transport chain (ETC) dysfunction due to mutations in the nuclear or mitochondrial genome is a common cause of metabolic disease in humans, and displays striking tissue specificity depending on the affected gene. The mechanisms underlying tissue specific phenotypes are not understood. Complex I (cI) is classically considered the entry point for electrons into the ETC, and in vitro experiments indicate that cI is required for basal respiration and maintenance of the NAD+/NADH ratio, an indicator of cellular redox status. This finding has largely not been tested in vivo. Here, we report that mitochondrial complex I (cI) is dispensable for homeostasis of the adult mouse liver; animals with hepatocyte-specific loss of cI function display no overt phenotypes or signs of liver damage, and maintain liver function, redox and oxygen status. Further analysis of cI-deficient livers did not reveal significant proteomic or metabolic changes, indicating little to no compensation is required in the setting of complex I loss. In contrast, complex IV (cIV) dysfunction in adult hepatocytes results in decreased liver function, impaired oxygen handling, steatosis, and liver damage, accompanied by significant metabolomic and proteomic perturbations. Metabolomic analysis suggests that the electron transfer flavoprotein complex constitutes a major route for electron entry into the hepatic ETC. Our results support a model whereby complex I loss is tolerated in the mouse liver because hepatocytes use alternative electron donors to fuel the mitochondrial ETC.","INSTITUTE":"The University of Texas Southwestern Medical Center at Dallas","DEPARTMENT":"Children's Research Institute","LABORATORY":"Prashant Mishra","LAST_NAME":"Lesner","FIRST_NAME":"Nicholas","ADDRESS":"6000 Harry Hines BLVD","EMAIL":"nicholas.lesner@pennmedicine.upenn.edu","PHONE":"2146483784"},

"STUDY":{"STUDY_TITLE":"Differential requirements for mitochondrial electron transport chain components in the adult murine liver - Lactate/Pyruvate Tolerance Test","STUDY_SUMMARY":"Hepatic Cox10 knockout mice were injected with a lactate/pyruvate (10:1) solution with 40% of the mixture labeled with [U-13C]. After 30 minutes, plasma and liver tissue was harvested. Metabolites were extracted, dried, and derivatized to form methoxime-TBDMS adducts.","INSTITUTE":"The University of Texas Southwestern Medical Center at Dallas","DEPARTMENT":"Children's Research Institute","LABORATORY":"Prashant Mishra","LAST_NAME":"Lesner","FIRST_NAME":"Nicholas","ADDRESS":"6000 Harry Hines BLVD","EMAIL":"nicholas.lesner@pennmedicine.upenn.edu","NUM_GROUPS":"2","TOTAL_SUBJECTS":"9","PHONE":"2146483784"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"263N",
"Factors":{"Genotype":"WT"},
"Additional sample data":{"Tissue":"Liver","RAW_FILE_NAME":"001"}
},
{
"Subject ID":"-",
"Sample ID":"263R",
"Factors":{"Genotype":"KO"},
"Additional sample data":{"Tissue":"Liver","RAW_FILE_NAME":"002"}
},
{
"Subject ID":"-",
"Sample ID":"264N",
"Factors":{"Genotype":"WT"},
"Additional sample data":{"Tissue":"Liver","RAW_FILE_NAME":"003"}
},
{
"Subject ID":"-",
"Sample ID":"264R",
"Factors":{"Genotype":"KO"},
"Additional sample data":{"Tissue":"Liver","RAW_FILE_NAME":"004"}
},
{
"Subject ID":"-",
"Sample ID":"264L",
"Factors":{"Genotype":"WT"},
"Additional sample data":{"Tissue":"Liver","RAW_FILE_NAME":"005"}
},
{
"Subject ID":"-",
"Sample ID":"268N",
"Factors":{"Genotype":"KO"},
"Additional sample data":{"Tissue":"Liver","RAW_FILE_NAME":"006"}
},
{
"Subject ID":"-",
"Sample ID":"283N",
"Factors":{"Genotype":"WT"},
"Additional sample data":{"Tissue":"Liver","RAW_FILE_NAME":"007"}
},
{
"Subject ID":"-",
"Sample ID":"283R",
"Factors":{"Genotype":"KO"},
"Additional sample data":{"Tissue":"Liver","RAW_FILE_NAME":"008"}
},
{
"Subject ID":"-",
"Sample ID":"283L",
"Factors":{"Genotype":"KO"},
"Additional sample data":{"Tissue":"Liver","RAW_FILE_NAME":"009"}
},
{
"Subject ID":"-",
"Sample ID":"263N_plasma",
"Factors":{"Genotype":"WT"},
"Additional sample data":{"Tissue":"Plasma","RAW_FILE_NAME":"010"}
},
{
"Subject ID":"-",
"Sample ID":"263R_plasma",
"Factors":{"Genotype":"KO"},
"Additional sample data":{"Tissue":"Plasma","RAW_FILE_NAME":"011"}
},
{
"Subject ID":"-",
"Sample ID":"264N_plasma",
"Factors":{"Genotype":"WT"},
"Additional sample data":{"Tissue":"Plasma","RAW_FILE_NAME":"012"}
},
{
"Subject ID":"-",
"Sample ID":"264R_plasma",
"Factors":{"Genotype":"KO"},
"Additional sample data":{"Tissue":"Plasma","RAW_FILE_NAME":"013"}
},
{
"Subject ID":"-",
"Sample ID":"264L_plasma",
"Factors":{"Genotype":"WT"},
"Additional sample data":{"Tissue":"Plasma","RAW_FILE_NAME":"014"}
},
{
"Subject ID":"-",
"Sample ID":"268N_plasma",
"Factors":{"Genotype":"KO"},
"Additional sample data":{"Tissue":"Plasma","RAW_FILE_NAME":"015"}
},
{
"Subject ID":"-",
"Sample ID":"283N_plasma",
"Factors":{"Genotype":"WT"},
"Additional sample data":{"Tissue":"Plasma","RAW_FILE_NAME":"016"}
},
{
"Subject ID":"-",
"Sample ID":"283R_plasma",
"Factors":{"Genotype":"KO"},
"Additional sample data":{"Tissue":"Plasma","RAW_FILE_NAME":"017"}
},
{
"Subject ID":"-",
"Sample ID":"283L_plasma",
"Factors":{"Genotype":"KO"},
"Additional sample data":{"Tissue":"Plasma","RAW_FILE_NAME":"018"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Liver was frozen on liqN2. Liver was crushed on liqN2, and metabolites extracted with 80% meOH. MeOH was dried, and samples derivatized to form methoxime-TBDMS adducts and immediately injected onto the GCMS.","SAMPLE_TYPE":"Liver"},

"TREATMENT":{"TREATMENT_SUMMARY":"Mice were IP injected with a lactate/pyruvate (10:1) solution."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"80% methanol containing metabolites was dried overnight in a speedvac. Dried pellets were derivatized using methoxyamine and MTBSTFA. Samples were immediately injected."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"60C for 1 min then 10C/min to 320C for 1 min 9 min at 320C","CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Agilent 7890B","COLUMN_NAME":"Agilent HP5-MS (30m x 0.25mm, 0.25 um)","INJECTION_TEMPERATURE":"60","INTERNAL_STANDARD":"Norvaline","SAMPLE_INJECTION":"Splitless"},

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

"MS":{"INSTRUMENT_NAME":"Agilent 5977A","INSTRUMENT_TYPE":"Single quadrupole","MS_TYPE":"EI","ION_MODE":"POSITIVE","MS_COMMENTS":"Scan 75 - 1000 m/z Data processed using Agilent Enhanced Data Analysis Peaks were auto integrated and confirmed by hand."},

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

"Data":[{"Metabolite":"Pyruvate","263N":"1041533","263R":"1138034","264N":"1067730","264R":"1273782","264L":"981941","268N":"1231814","283N":"1250459","283R":"1497066","283L":"1187086"},{"Metabolite":"aHB","263N":"151799","263R":"91533","264N":"220056","264R":"187999","264L":"185179","268N":"131451","283N":"89343","283R":"181646","283L":"120847"},{"Metabolite":"bHB","263N":"958969","263R":"711112","264N":"2394679","264R":"1919596","264L":"1828968","268N":"1120091","283N":"844942","283R":"230819","283L":"769859"},{"Metabolite":"Lactate","263N":"10164430","263R":"14041435","264N":"13316188","264R":"16139552","264L":"10685685","268N":"14807413","283N":"11444081","283R":"17769939","283L":"13282912"},{"Metabolite":"Alanine","263N":"93241","263R":"250131","264N":"104769","264R":"168099","264L":"115372","268N":"211150","283N":"57855","283R":"630535","283L":"233519"},{"Metabolite":"Glycine","263N":"18153","263R":"18694","264N":"10890","264R":"2594546","264L":"15946","268N":"17563","283N":"9111","283R":"29265","283L":"20760"},{"Metabolite":"Valine","263N":"110323","263R":"136328","264N":"97432","264R":"102356","264L":"106208","268N":"171896","283N":"48644","283R":"96311","283L":"86982"},{"Metabolite":"Leucine","263N":"179742","263R":"247752","264N":"182382","264R":"171495","264L":"185704","268N":"324437","283N":"73933","283R":"154255","283L":"119463"},{"Metabolite":"Isoleucine","263N":"93786","263R":"148407","264N":"104267","264R":"115868","264L":"83342","268N":"125842","283N":"53395","283R":"102348","283L":"96487"},{"Metabolite":"Proline","263N":"12627","263R":"27503","264N":"15217","264R":"22709","264L":"20506","268N":"25742","283N":"9361","283R":"45174","283L":"26472"},{"Metabolite":"Succinate","263N":"5134","263R":"5670","264N":"6764","264R":"11849","264L":"3488","268N":"4873","283N":"2735","283R":"3984","283L":"3172"},{"Metabolite":"Fumarate","263N":"5686","263R":"8111","264N":"6996","264R":"6975","264L":"5349","268N":"4910","283N":"3460","283R":"5640","283L":"5925"},{"Metabolite":"Methionine","263N":"21027","263R":"55715","264N":"26723","264R":"40680","264L":"29732","268N":"56792","283N":"21125","283R":"73378","283L":"69464"},{"Metabolite":"Serine","263N":"37012","263R":"87490","264N":"48970","264R":"75153","264L":"50484","268N":"81391","283N":"25407","283R":"109959","283L":"93136"},{"Metabolite":"Threonine","263N":"138178","263R":"319698","264N":"147531","264R":"230199","264L":"201786","268N":"315524","283N":"96853","283R":"333346","283L":"367927"},{"Metabolite":"Phenylalanine","263N":"89034","263R":"141147","264N":"107834","264R":"134074","264L":"106583","268N":"165309","283N":"65127","283R":"123932","283L":"102596"},{"Metabolite":"aKG","263N":"728","263R":"927","264N":"769","264R":"777","264L":"977","268N":"725","283N":"921","283R":"1335","283L":"1514"},{"Metabolite":"Malate","263N":"0","263R":"0","264N":"0","264R":"0","264L":"0","268N":"513","283N":"0","283R":"686","283L":"691"},{"Metabolite":"Aspartate","263N":"0","263R":"0","264N":"0","264R":"0","264L":"0","268N":"0","283N":"0","283R":"0","283L":"0"},{"Metabolite":"Cysteine","263N":"4278","263R":"6208","264N":"7312","264R":"1024","264L":"7960","268N":"6910","283N":"5404","283R":"10147","283L":"9226"},{"Metabolite":"Glutamate","263N":"0","263R":"2094","264N":"0","264R":"1494","264L":"908","268N":"0","283N":"518","283R":"1764","283L":"1963"},{"Metabolite":"Lysine","263N":"0","263R":"989","264N":"0","264R":"583","264L":"0","268N":"1006","283N":"0","283R":"2317","283L":"1162"},{"Metabolite":"Glutamine","263N":"6841","263R":"15836","264N":"11490","264R":"16867","264L":"14052","268N":"15419","283N":"6536","283R":"97230","283L":"23672"},{"Metabolite":"Histidine","263N":"1915","263R":"6025","264N":"3049","264R":"7581","264L":"2423","268N":"6905","283N":"1545","283R":"20878","283L":"7360"},{"Metabolite":"Citrate","263N":"19161","263R":"14235","264N":"16351","264R":"19980","264L":"19102","268N":"15940","283N":"19890","283R":"15141","283L":"17850"},{"Metabolite":"3PG","263N":"0","263R":"0","264N":"0","264R":"0","264L":"0","268N":"0","283N":"0","283R":"0","283L":"0"},{"Metabolite":"Tyrosine","263N":"30601","263R":"93965","264N":"26446","264R":"70766","264L":"36855","268N":"104389","283N":"18459","283R":"199882","283L":"75573"},{"Metabolite":"PEP","263N":"0","263R":"0","264N":"0","264R":"0","264L":"0","268N":"0","283N":"0","283R":"0","283L":"0"},{"Metabolite":"Glucose","263N":"540177","263R":"476708","264N":"525141","264R":"530942","264L":"513437","268N":"493169","283N":"521420","283R":"386827","283L":"510752"},{"Metabolite":"d4-BHB","263N":"1553725","263R":"1624220","264N":"1667270","264R":"1616022","264L":"1443093","268N":"1442617","283N":"1499418","283R":"1048560","283L":"1503391"},{"Metabolite":"aKB","263N":"260020","263R":"134525","264N":"258317","264R":"156714","264L":"267733","268N":"184986","283N":"163746","283R":"181276","283L":"201509"}],

"Metabolites":[{"metabolite_name":"Pyruvate"},{"metabolite_name":"aHB"},{"metabolite_name":"bHB"},{"metabolite_name":"Lactate"},{"metabolite_name":"Alanine"},{"metabolite_name":"Glycine"},{"metabolite_name":"Valine"},{"metabolite_name":"Leucine"},{"metabolite_name":"Isoleucine"},{"metabolite_name":"Proline"},{"metabolite_name":"Succinate"},{"metabolite_name":"Fumarate"},{"metabolite_name":"Methionine"},{"metabolite_name":"Serine"},{"metabolite_name":"Threonine"},{"metabolite_name":"Phenylalanine"},{"metabolite_name":"aKG"},{"metabolite_name":"Malate"},{"metabolite_name":"Aspartate"},{"metabolite_name":"Cysteine"},{"metabolite_name":"Glutamate"},{"metabolite_name":"Lysine"},{"metabolite_name":"Glutamine"},{"metabolite_name":"Histidine"},{"metabolite_name":"Citrate"},{"metabolite_name":"3PG"},{"metabolite_name":"Tyrosine"},{"metabolite_name":"PEP"},{"metabolite_name":"Glucose"},{"metabolite_name":"d4-BHB"},{"metabolite_name":"aKB"}]
}

}