{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003112","ANALYSIS_ID":"AN005111","VERSION":"1","CREATED_ON":"March 7, 2024, 1:02 pm"},

"PROJECT":{"PROJECT_TITLE":"Glucose Hypometabolism Prompts RAN Translation and Exacerbates C9orf72-related ALS/FTD Phenotypes","PROJECT_SUMMARY":"The most prevalent genetic cause of both amyotrophic lateral sclerosis and frontotemporal dementia is a (GGGGCC)n nucleotide repeat expansion (NRE) occurring in the first intron of the C9orf72 gene (C9). Brain glucose hypometabolism is consistently observed in C9-NRE carriers, even at pre-symptomatic stages, although its potential role in disease pathogenesis is unknown. Here, we identified alterations in glucose metabolic pathways and ATP levels in the brains of asymptomatic C9-BAC mice. We found that, through activation of the GCN2 kinase, glucose hypometabolism drives the production of dipeptide repeat proteins (DPRs), impairs the survival of C9 patient-derived neurons, and triggers motor dysfunction in C9-BAC mice. We also found that one of the arginine-rich DPRs (PR) can directly contribute to glucose metabolism and metabolic stress. These findings provide a potential mechanistic link between energy imbalances and C9-ALS/FTD pathogenesis and suggest a feedforward loop model that opens several opportunities for therapeutic intervention.","INSTITUTE":"Thomas Jefferson University","LAST_NAME":"Davide","FIRST_NAME":"Trotti","ADDRESS":"900 Walnut Street, Philadelphia, PA 19107, USA","EMAIL":"davide.trotti@jefferson.edu","PHONE":"215-955-8416"},

"STUDY":{"STUDY_TITLE":"Glucose Hypometabolism Prompts RAN Translation and Exacerbates C9orf72-related ALS/FTD Phenotypes","STUDY_SUMMARY":"The most prevalent genetic cause of both amyotrophic lateral sclerosis and frontotemporal dementia is a (GGGGCC)n nucleotide repeat expansion (NRE) occurring in the first intron of the C9orf72 gene (C9). Brain glucose hypometabolism is consistently observed in C9-NRE carriers, even at pre-symptomatic stages, although its potential role in disease pathogenesis is unknown. Here, we identified alterations in glucose metabolic pathways and ATP levels in the brains of asymptomatic C9-BAC mice. We found that, through activation of the GCN2 kinase, glucose hypometabolism drives the production of dipeptide repeat proteins (DPRs), impairs the survival of C9 patient-derived neurons, and triggers motor dysfunction in C9-BAC mice. We also found that one of the arginine-rich DPRs (PR) can directly contribute to glucose metabolism and metabolic stress. These findings provide a potential mechanistic link between energy imbalances and C9-ALS/FTD pathogenesis and suggest a feedforward loop model that opens several opportunities for therapeutic intervention.","INSTITUTE":"Thomas Jefferson University","LAST_NAME":"Trotti","FIRST_NAME":"Davide","ADDRESS":"900 Walnut Street, Philadelphia, PA 19107, USA","EMAIL":"davide.trotti@jefferson.edu","PHONE":"215-955-8416"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"C92DG_07",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"2DG"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C92DG_07.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C92DG_26",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"2DG"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C92DG_26.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C92DG_30",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"2DG"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C92DG_30.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C92DG_34",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"2DG"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C92DG_34.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C92DG_45",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"2DG"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C92DG_45.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C92DG_47",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"2DG"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C92DG_47.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C92DG_56",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"2DG"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C92DG_56.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C9S_11",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C9S_11.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C9S_29",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C9S_29.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C9S_33",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C9S_33.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C9S_43",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C9S_43.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C9S_50",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C9S_50.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C9S_57",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C9S_57.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"C9S_69",
"Factors":{"Sample source":"Brain","Genotype":"C9orf72","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_C9S_69.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"WTS_31",
"Factors":{"Sample source":"Brain","Genotype":"WT","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_WTS_31.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"WTS_39",
"Factors":{"Sample source":"Brain","Genotype":"WT","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_WTS_39.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"WTS_51",
"Factors":{"Sample source":"Brain","Genotype":"WT","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_WTS_51.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"WTS_53",
"Factors":{"Sample source":"Brain","Genotype":"WT","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_WTS_53.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"WTS_58",
"Factors":{"Sample source":"Brain","Genotype":"WT","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_WTS_58.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"WTS_59",
"Factors":{"Sample source":"Brain","Genotype":"WT","Treatment":"Saline"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME(Raw file name)":"HF1170_WTS_59.mzXML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Animals were euthanized by CO2 inhalation 16 hours following the final injection and transcardially perfused with ice-cold PBS. Whole brains and spinal cords were then excised, flash frozen in liquid N2, and stored at –80°C","SAMPLE_TYPE":"Brain cortex"},

"TREATMENT":{"TREATMENT_SUMMARY":"Animals were administered i.p. injections of saline or4 g/kg 2DG twice weekly for six weeks"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Metabolites from equal amounts of frontal cortex tissue were rapidly extracted in 80% ice-cold methanol. Extracted samples were vortexed twice, cleared by centrifugation at 14,000 x g for 20 minutes at 4°C, and stored at -80°C."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um)","SOLVENT_A":"100% acetonitrile","SOLVENT_B":"0.1% NH4OH/20 mM CH3COONH4 in water","FLOW_GRADIENT":"85% to 30% A in 20 min followed by a wash with 30% A and re-equilibration at 85% A","FLOW_RATE":"150 μL/min","COLUMN_TEMPERATURE":"30"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"UNSPECIFIED","MS_COMMENTS":"A Sequant ZIC-pHILIC column (2.1 mm i.d. × 150 mm, particle size of 5 µm, Millipore Sigma) was used for separation of metabolites. A 2.1 × 20 mm guard column with the same packing material was used for protection of the analytical column. Flow rate was set at 150 μL/min. Buffers consisted of 100% acetonitrile for mobile phase A, and 0.1% NH4OH/20 mM CH3COONH4 in water for mobile phase B. The chromatographic gradient ran from 85% to 30% A in 20 min followed by a wash with 30% A and re-equilibration at 85% A. Column temperature was 30 C."},

"MS_METABOLITE_DATA":{
"Units":"peak intensity",

"Data":[{"Metabolite":"1-Methyladenosine","C92DG_07":"32617.51","C92DG_26":"13589.21","C92DG_30":"17550.16","C92DG_34":"14579.14","C92DG_45":"24581.4","C92DG_47":"10393.55","C92DG_56":"23581.61","C9S_11":"17135.79","C9S_29":"6183.92","C9S_33":"3205.882","C9S_43":"22935.13","C9S_50":"30999.28","C9S_57":"14765.45","C9S_69":"21999.67","WTS_31":"20756.85","WTS_39":"14962.61","WTS_51":"10067.7","WTS_53":"9564.473","WTS_58":"13554.18","WTS_59":"15764.18"},{"Metabolite":"1-Methylnicotinamide","C92DG_07":"1841007","C92DG_26":"1822386","C92DG_30":"2718988","C92DG_34":"4048276","C92DG_45":"688253.6","C92DG_47":"1131924","C92DG_56":"1370287","C9S_11":"2228298","C9S_29":"1856145","C9S_33":"2501936","C9S_43":"1516392","C9S_50":"3333350","C9S_57":"2220781","C9S_69":"2493598","WTS_31":"863027","WTS_39":"1715365","WTS_51":"1386362","WTS_53":"1732681","WTS_58":"2033457","WTS_59":"2404083"},{"Metabolite":"2DG","C92DG_07":"0","C92DG_26":"0","C92DG_30":"0","C92DG_34":"0","C92DG_45":"0","C92DG_47":"0","C92DG_56":"0","C9S_11":"0","C9S_29":"0","C9S_33":"0","C9S_43":"0","C9S_50":"0","C9S_57":"0","C9S_69":"0","WTS_31":"0","WTS_39":"0","WTS_51":"0","WTS_53":"0","WTS_58":"0","WTS_59":"0"},{"Metabolite":"2DGP","C92DG_07":"1142208","C92DG_26":"5765284","C92DG_30":"3171172","C92DG_34":"3409532","C92DG_45":"3425847","C92DG_47":"2534675","C92DG_56":"4221022","C9S_11":"0","C9S_29":"79711.85","C9S_33":"84458.56","C9S_43":"609323.9","C9S_50":"100214.5","C9S_57":"91255.3","C9S_69":"98635.8","WTS_31":"322796.1","WTS_39":"39111.83","WTS_51":"334947.6","WTS_53":"127164.3","WTS_58":"225007.6","WTS_59":"222710.2"},{"Metabolite":"2-Hydroxycaproic acid","C92DG_07":"1493785","C92DG_26":"738976.2","C92DG_30":"787589.1","C92DG_34":"882673.2","C92DG_45":"878327.1","C92DG_47":"1072232","C92DG_56":"473638.9","C9S_11":"723708.3","C9S_29":"503513.9","C9S_33":"413687.8","C9S_43":"428152.3","C9S_50":"881504","C9S_57":"442938.1","C9S_69":"581325.1","WTS_31":"1030926","WTS_39":"558206.3","WTS_51":"988875.9","WTS_53":"546005.7","WTS_58":"819152.1","WTS_59":"948766"},{"Metabolite":"2-Hydroxyvaleric acid","C92DG_07":"1786160","C92DG_26":"2013362","C92DG_30":"1812120","C92DG_34":"2281630","C92DG_45":"1299487","C92DG_47":"1826766","C92DG_56":"2240205","C9S_11":"2265580","C9S_29":"1460612","C9S_33":"701437.8","C9S_43":"1634673","C9S_50":"3061003","C9S_57":"2739082","C9S_69":"1760775","WTS_31":"1199602","WTS_39":"1817412","WTS_51":"2131307","WTS_53":"1898601","WTS_58":"2717337","WTS_59":"1795729"},{"Metabolite":"3-Hydroxymethylglutaric acid","C92DG_07":"6871342","C92DG_26":"7208425","C92DG_30":"7909773","C92DG_34":"8807470","C92DG_45":"3802989","C92DG_47":"5161551","C92DG_56":"5561114","C9S_11":"5613614","C9S_29":"5979359","C9S_33":"4723424","C9S_43":"4886748","C9S_50":"6777701","C9S_57":"5942222","C9S_69":"4827469","WTS_31":"4337369","WTS_39":"5899910","WTS_51":"5544885","WTS_53":"6513512","WTS_58":"7142908","WTS_59":"9241531"},{"Metabolite":"3-Indolepropionic acid","C92DG_07":"6480.795","C92DG_26":"4296.553","C92DG_30":"4321.637","C92DG_34":"8103.824","C92DG_45":"3463.247","C92DG_47":"7244.678","C92DG_56":"6293.21","C9S_11":"8503.848","C9S_29":"5235.161","C9S_33":"2868.246","C9S_43":"4797.81","C9S_50":"5596.526","C9S_57":"5175.813","C9S_69":"3256.446","WTS_31":"2974.045","WTS_39":"6200.765","WTS_51":"3855.611","WTS_53":"7870.383","WTS_58":"4325.25","WTS_59":"7757.958"},{"Metabolite":"3-Methylhistidine","C92DG_07":"3209961","C92DG_26":"5886811","C92DG_30":"4186838","C92DG_34":"4295106","C92DG_45":"2501087","C92DG_47":"4280228","C92DG_56":"5209526","C9S_11":"4254567","C9S_29":"4983342","C9S_33":"2131721","C9S_43":"2978088","C9S_50":"7089334","C9S_57":"6117056","C9S_69":"5205673","WTS_31":"3390343","WTS_39":"3253151","WTS_51":"6593800","WTS_53":"5011118","WTS_58":"9165029","WTS_59":"7773793"},{"Metabolite":"3-Phosphoglyceric acid","C92DG_07":"292058.1","C92DG_26":"374590.8","C92DG_30":"224060.3","C92DG_34":"369201","C92DG_45":"309884.6","C92DG_47":"662231.3","C92DG_56":"410955.5","C9S_11":"465306","C9S_29":"284887.6","C9S_33":"107345.5","C9S_43":"288491.3","C9S_50":"470891.8","C9S_57":"464510.8","C9S_69":"166406.9","WTS_31":"383866.4","WTS_39":"349232.5","WTS_51":"486476.7","WTS_53":"405851.3","WTS_58":"430072.2","WTS_59":"691187.1"},{"Metabolite":"4-Acetamidobutanoic acid","C92DG_07":"0","C92DG_26":"0","C92DG_30":"0","C92DG_34":"0","C92DG_45":"0","C92DG_47":"0","C92DG_56":"0","C9S_11":"0","C9S_29":"0","C9S_33":"0","C9S_43":"0","C9S_50":"0","C9S_57":"0","C9S_69":"0","WTS_31":"0","WTS_39":"0","WTS_51":"0","WTS_53":"0","WTS_58":"0","WTS_59":"0"},{"Metabolite":"4-Hydroxyproline","C92DG_07":"0","C92DG_26":"0","C92DG_30":"0","C92DG_34":"0","C92DG_45":"0","C92DG_47":"0","C92DG_56":"0","C9S_11":"0","C9S_29":"0","C9S_33":"0","C9S_43":"0","C9S_50":"0","C9S_57":"0","C9S_69":"0","WTS_31":"0","WTS_39":"0","WTS_51":"0","WTS_53":"0","WTS_58":"0","WTS_59":"0"},{"Metabolite":"4-Pyridoxic acid","C92DG_07":"250103.5","C92DG_26":"208616.3","C92DG_30":"196308","C92DG_34":"217139.4","C92DG_45":"194310.2","C92DG_47":"230841","C92DG_56":"180077.5","C9S_11":"208649","C9S_29":"171939.6","C9S_33":"115284","C9S_43":"136693.4","C9S_50":"310870.8","C9S_57":"190266.8","C9S_69":"156890.7","WTS_31":"189076.4","WTS_39":"216055.6","WTS_51":"221708.2","WTS_53":"221993.8","WTS_58":"245587.3","WTS_59":"210657.7"},{"Metabolite":"4-Trimethylammoniobutanoic acid","C92DG_07":"5444141","C92DG_26":"15619190","C92DG_30":"13888740","C92DG_34":"16743310","C92DG_45":"5397229","C92DG_47":"15800820","C92DG_56":"7550785","C9S_11":"7648567","C9S_29":"10546330","C9S_33":"5213951","C9S_43":"4841650","C9S_50":"5249534","C9S_57":"6472307","C9S_69":"4263275","WTS_31":"6027153","WTS_39":"11679830","WTS_51":"10448560","WTS_53":"6236550","WTS_58":"8219130","WTS_59":"3749197"},{"Metabolite":"5-Thymidylic acid","C92DG_07":"0","C92DG_26":"0","C92DG_30":"0","C92DG_34":"0","C92DG_45":"0","C92DG_47":"0","C92DG_56":"0","C9S_11":"0","C9S_29":"0","C9S_33":"0","C9S_43":"0","C9S_50":"0","C9S_57":"0","C9S_69":"0","WTS_31":"0","WTS_39":"0","WTS_51":"0","WTS_53":"0","WTS_58":"0","WTS_59":"0"},{"Metabolite":"6-Phosphogluconic 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}

}