{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001819","ANALYSIS_ID":"AN002953","VERSION":"1","CREATED_ON":"June 7, 2021, 12:24 pm"},

"PROJECT":{"PROJECT_TYPE":"Lipidomics analysis","PROJECT_SUMMARY":"Apolipoprotein E ε4 (APOE4) is the primary genetic risk factor for the late-onset form of Alzheimer's disease (AD). Although the reason for this association is not completely understood, researchers have uncovered numerous effects of APOE4 expression on AD-relevant brain processes, including amyloid beta (Aβ) accumulation, lipid metabolism, endosomal-lysosomal processing and bioenergetics. In this study, we aimed to determine the effect of APOE4 allelic dosage on regional brain lipid composition in aged mice, as well as in cultured neurons. We performed a targeted lipidomic analysis on the entorhinal cortex (EC) and primary visual cortex (PVC) from 14–15 month-old APOE3/3, APOE3/4, and APOE4/4 targeted replacement mice, as well as on WT neurons cultured with conditioned media from APOE3/3 or APOE4/4 astrocytes. Our results reveal that the EC possesses increased susceptibility to APOE4-associated lipid alterations compared to the PVC. In the EC, APOE4 expression showed a dominant effect in decreasing diacylglycerol (DAG) levels, and a semi-dominant additive effect in the upregulation of multiple ceramide, glycosylated sphingolipid and bis(monoacylglycerol)phosphate (BMP) species, lipids known to accumulate as a result of endosomal-lysosomal dysfunction and defective lysosomal clearance. Neurons treated with conditioned media from APOE4 vs. APOE3 astrocytes also showed similar alterations of DAG and BMP species as those observed in the mouse EC. Our results suggest that APOE4 expression differentially modulates regional and neuronal lipid signatures, which may underlie the increased susceptibility of EC-localized neurons to AD pathology.","INSTITUTE":"Columbia University","FIRST_NAME":"Tal","ADDRESS":"630 W 168th St., P&S 12-430","EMAIL":"tn2283@cumc.columbia.edu","PHONE":"2123045683","PROJECT_TITLE":"APOE4-associated differences in lipidomics signatures in mouse brain and cultured neurons","LAST_NAME":"Nuriel"},

"STUDY":{"STUDY_TITLE":"EC and PVC from 14-15 month-old APOE3/3, APOE3/4 and APOE4/4 mice","STUDY_SUMMARY":"We performed a targeted lipidomic analysis on EC and PVC tissue from 14-15 month old APOE3/3, APOE3/4, and APOE4/4 mice.","INSTITUTE":"Columbia University","LAST_NAME":"Nuriel","FIRST_NAME":"Tal","ADDRESS":"630 W 168th St., P&S 12-430","EMAIL":"tn2283@cumc.columbia.edu","PHONE":"2123045683"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","AGE_OR_AGE_RANGE":"14-15 month-old","GENDER":"Male"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"EC1",
"Factors":{"Region":"EC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"EC2",
"Factors":{"Region":"EC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"EC3",
"Factors":{"Region":"EC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"EC4",
"Factors":{"Region":"EC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"EC5",
"Factors":{"Region":"EC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"EC6",
"Factors":{"Region":"EC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"EC7",
"Factors":{"Region":"EC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"EC8",
"Factors":{"Region":"EC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"EC10",
"Factors":{"Region":"EC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC12",
"Factors":{"Region":"EC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC13",
"Factors":{"Region":"EC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC14",
"Factors":{"Region":"EC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC15",
"Factors":{"Region":"EC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC16",
"Factors":{"Region":"EC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC17",
"Factors":{"Region":"EC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC18",
"Factors":{"Region":"EC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC19",
"Factors":{"Region":"EC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC20",
"Factors":{"Region":"EC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC21",
"Factors":{"Region":"EC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC22",
"Factors":{"Region":"EC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC23",
"Factors":{"Region":"EC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"EC24",
"Factors":{"Region":"EC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC1",
"Factors":{"Region":"PVC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"PVC2",
"Factors":{"Region":"PVC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"PVC3",
"Factors":{"Region":"PVC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"PVC4",
"Factors":{"Region":"PVC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"PVC5",
"Factors":{"Region":"PVC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"PVC6",
"Factors":{"Region":"PVC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"PVC7",
"Factors":{"Region":"PVC","Genotype":"E3/3"}
},
{
"Subject ID":"-",
"Sample ID":"PVC9",
"Factors":{"Region":"PVC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC10",
"Factors":{"Region":"PVC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC11",
"Factors":{"Region":"PVC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC12",
"Factors":{"Region":"PVC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC13",
"Factors":{"Region":"PVC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC14",
"Factors":{"Region":"PVC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC15",
"Factors":{"Region":"PVC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC16",
"Factors":{"Region":"PVC","Genotype":"E3/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC17",
"Factors":{"Region":"PVC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC19",
"Factors":{"Region":"PVC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC20",
"Factors":{"Region":"PVC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC21",
"Factors":{"Region":"PVC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC22",
"Factors":{"Region":"PVC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC23",
"Factors":{"Region":"PVC","Genotype":"E4/4"}
},
{
"Subject ID":"-",
"Sample ID":"PVC24",
"Factors":{"Region":"PVC","Genotype":"E4/4"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Mice were sacrificed by cervical dislocation to maintain the brain environment, and individual brain regions were immediately removed and snap-frozen on dry ice. Tissues were stored at -80°C for prior to extraction.","SAMPLE_TYPE":"Brain"},

"TREATMENT":{"TREATMENT_SUMMARY":"No treatments were made. Mice were either APOE3/3, APOE3/4 or APOE4/4 gentoype."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Lipid and small-molecule metabolite extraction was performed using a methyl tert-butyl ether (MTBE)/methanol extraction protocol modified from previous reports (40, 41), as we have described previously (34). Briefly, individual EC or PVC tissues were homogenized in 400 ul of ice-cold methanol using a bead mill homogenizer (TissueLyser II, Qiagen) at 25 beats/sec, 2x for 45 sec each. Following homogenization, samples were incubated in 1200 ul of MTBE for 1 hr at room temperature to separate organic-soluble lipids from aqueous-soluble lipids and other small-molecules. Finally, 360 ul of ultrapure water was added (for a final ratio of 3:1:0.9 MTBE:methanol:water) to resolve the two liquid phases, and each samples were centrifuged at 10,000 x g for 10 min. For this experiment, the upper organic phase was collected from each sample and stored in a separate tube, and the remaining protein pellets were resuspended in 25 mM ammonium bicarbonate, pH 8, with 2.5% SDS. A BCA protein assay was performed on each protein fraction, and the organic phase was normalized to their protein concentration equivalent with 100% methanol. All samples were then stored at -80°C prior to analysis."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Normal phase","CHROMATOGRAPHY_TYPE":"Normal phase","INSTRUMENT_NAME":"Agilent 1260","COLUMN_NAME":"Phenomenex Luna NH2 (150 x 2.1mm, 3um)"},

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

"MS":{"INSTRUMENT_NAME":"Agilent 6490 QQQ","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Lipid profiling was performed using an Agilent 1260 HPLC coupled to an Agilent 6490 triple quadrupole (QQQ) mass spectrometer (41). Each sample was run through three separate chromatographic conditions (reverse-phase negative mode, reverse-phase positive mode and normal-phase positive mode) for the effective quantification of 337 distinct lipids from 28 lipid subclasses, as previously described (41). Individual lipid species were measured by multiple reaction monitoring transitions."},

"MS_METABOLITE_DATA":{
"Units":"area under the curve",

"Data":[{"Metabolite":"FC","EC1":"9492.3038","EC2":"10305.0996","EC3":"11709.4660","EC4":"10290.5312","EC5":"9825.1637","EC6":"7660.4514","EC7":"8244.9902","EC8":"8971.0081","EC10":"10862.5112","EC12":"9719.2992","EC13":"8937.4274","EC14":"9479.0168","EC15":"9064.6821","EC16":"9011.9674","EC17":"11347.1270","EC18":"9073.4543","EC19":"10025.7376","EC20":"11074.5775","EC21":"9357.4495","EC22":"11086.0010","EC23":"9173.8179","EC24":"10543.9640","PVC1":"9310.2035","PVC2":"8787.7746","PVC3":"7978.3804","PVC4":"8217.9342","PVC5":"7981.8396","PVC6":"7807.8556","PVC7":"8072.3952","PVC9":"9352.2577","PVC10":"8771.9253","PVC11":"7247.5313","PVC12":"8588.9387","PVC13":"8963.6479","PVC14":"9668.9699","PVC15":"8335.8507","PVC16":"8533.0630","PVC17":"6875.7376","PVC19":"9091.6383","PVC20":"9356.1407","PVC21":"8244.8213","PVC22":"9111.3913","PVC23":"8918.4002","PVC24":"8499.4431"},{"Metabolite":"Chol 24-OH","EC1":"0.2487","EC2":"0.2733","EC3":"0.3937","EC4":"0.2838","EC5":"0.4519","EC6":"0.3721","EC7":"0.4376","EC8":"0.2778","EC10":"0.3334","EC12":"0.6273","EC13":"0.4474","EC14":"0.7942","EC15":"0.4742","EC16":"0.4078","EC17":"0.3670","EC18":"0.4741","EC19":"0.3695","EC20":"0.5196","EC21":"0.4995","EC22":"0.5158","EC23":"0.5390","EC24":"0.2961","PVC1":"0.2978","PVC2":"0.4603","PVC3":"0.2982","PVC4":"0.4126","PVC5":"0.2314","PVC6":"0.4693","PVC7":"0.3984","PVC9":"0.2224","PVC10":"0.6241","PVC11":"0.4933","PVC12":"0.3333","PVC13":"0.4109","PVC14":"0.4265","PVC15":"0.4853","PVC16":"0.3053","PVC17":"0.2747","PVC19":"0.3945","PVC20":"0.2752","PVC21":"0.2716","PVC22":"0.4750","PVC23":"0.3759","PVC24":"0.2830"},{"Metabolite":"Chol 27-OH","EC1":"0.2681","EC2":"0.3516","EC3":"0.3600","EC4":"0.2648","EC5":"0.5512","EC6":"0.5197","EC7":"0.5144","EC8":"0.3316","EC10":"0.3347","EC12":"0.6727","EC13":"0.6069","EC14":"0.8971","EC15":"0.5063","EC16":"0.3786","EC17":"0.3355","EC18":"0.3486","EC19":"0.7951","EC20":"0.4520","EC21":"0.8269","EC22":"0.5952","EC23":"0.5450","EC24":"0.3569","PVC1":"0.3454","PVC2":"0.4119","PVC3":"0.3435","PVC4":"0.3886","PVC5":"0.2612","PVC6":"0.3462","PVC7":"0.3465","PVC9":"0.2464","PVC10":"0.5274","PVC11":"0.3071","PVC12":"0.2325","PVC13":"0.3774","PVC14":"0.5053","PVC15":"0.3986","PVC16":"0.2649","PVC17":"0.2610","PVC19":"0.3349","PVC20":"0.2866","PVC21":"0.2817","PVC22":"0.5527","PVC23":"0.3990","PVC24":"0.2895"},{"Metabolite":"CE 16:0","EC1":"0.0136","EC2":"0.0130","EC3":"0.0074","EC4":"0.0184","EC5":"0.0196","EC6":"0.0222","EC7":"0.0451","EC8":"0.0106","EC10":"0.0093","EC12":"0.0093","EC13":"0.0168","EC14":"0.0111","EC15":"0.0157","EC16":"0.0096","EC17":"0.0317","EC18":"0.0168","EC19":"0.0103","EC20":"0.0173","EC21":"0.0109","EC22":"0.0193","EC23":"0.0174","EC24":"0.0118","PVC1":"0.0135","PVC2":"0.0112","PVC3":"0.0123","PVC4":"0.0077","PVC5":"0.0136","PVC6":"0.0068","PVC7":"0.0142","PVC9":"0.0100","PVC10":"0.1214","PVC11":"0.0107","PVC12":"0.0110","PVC13":"0.0109","PVC14":"0.0121","PVC15":"0.0100","PVC16":"0.0107","PVC17":"0.0071","PVC19":"0.0124","PVC20":"0.0144","PVC21":"0.0039","PVC22":"0.0174","PVC23":"0.0544","PVC24":"0.0094"},{"Metabolite":"CE 16:1","EC1":"0.0192","EC2":"0.0206","EC3":"0.0262","EC4":"0.0213","EC5":"0.0267","EC6":"0.0438","EC7":"0.1365","EC8":"0.0209","EC10":"0.0247","EC12":"0.0155","EC13":"0.0311","EC14":"0.0165","EC15":"0.0486","EC16":"0.0159","EC17":"0.0338","EC18":"0.0432","EC19":"0.0248","EC20":"0.0420","EC21":"0.0126","EC22":"0.0406","EC23":"0.0326","EC24":"0.0266","PVC1":"0.0303","PVC2":"0.0170","PVC3":"0.0184","PVC4":"0.0138","PVC5":"0.0197","PVC6":"0.0166","PVC7":"0.0314","PVC9":"0.0258","PVC10":"0.0817","PVC11":"0.0139","PVC12":"0.0109","PVC13":"0.0203","PVC14":"0.0306","PVC15":"0.0176","PVC16":"0.0142","PVC17":"0.0152","PVC19":"0.0244","PVC20":"0.0255","PVC21":"0.0129","PVC22":"0.0354","PVC23":"0.0573","PVC24":"0.0140"},{"Metabolite":"CE 18:0","EC1":"0.0067","EC2":"0.0063","EC3":"0.0103","EC4":"0.0047","EC5":"0.0072","EC6":"0.0070","EC7":"0.0119","EC8":"0.0075","EC10":"0.0056","EC12":"0.0059","EC13":"0.0050","EC14":"0.0062","EC15":"0.0064","EC16":"0.0032","EC17":"0.0108","EC18":"0.0051","EC19":"0.0047","EC20":"0.0050","EC21":"0.0069","EC22":"0.0043","EC23":"0.0057","EC24":"0.0062","PVC1":"0.0066","PVC2":"0.0070","PVC3":"0.0031","PVC4":"0.0053","PVC5":"0.0051","PVC6":"0.0045","PVC7":"0.0040","PVC9":"0.0043","PVC10":"0.0215","PVC11":"0.0058","PVC12":"0.0048","PVC13":"0.0039","PVC14":"0.0043","PVC15":"0.0017","PVC16":"0.0049","PVC17":"0.0034","PVC19":"0.0046","PVC20":"0.0057","PVC21":"0.0049","PVC22":"0.0069","PVC23":"0.0127","PVC24":"0.0043"},{"Metabolite":"CE 18:1","EC1":"0.0548","EC2":"0.0692","EC3":"0.0562","EC4":"0.0873","EC5":"0.0813","EC6":"0.1312","EC7":"0.3855","EC8":"0.0707","EC10":"0.0572","EC12":"0.0526","EC13":"0.0640","EC14":"0.0425","EC15":"0.1191","EC16":"0.0507","EC17":"0.2386","EC18":"0.1040","EC19":"0.0692","EC20":"0.0929","EC21":"0.0472","EC22":"0.1038","EC23":"0.0784","EC24":"0.0837","PVC1":"0.0645","PVC2":"0.0731","PVC3":"0.0491","PVC4":"0.0507","PVC5":"0.0607","PVC6":"0.0418","PVC7":"0.0514","PVC9":"0.0575","PVC10":"0.3337","PVC11":"0.0288","PVC12":"0.0399","PVC13":"0.0549","PVC14":"0.0695","PVC15":"0.0449","PVC16":"0.0777","PVC17":"0.0459","PVC19":"0.1075","PVC20":"0.0902","PVC21":"0.0299","PVC22":"0.1038","PVC23":"0.2004","PVC24":"0.0487"},{"Metabolite":"CE 18:2","EC1":"0.3775","EC2":"0.4616","EC3":"0.5271","EC4":"0.3294","EC5":"0.6635","EC6":"0.6015","EC7":"1.9213","EC8":"0.8023","EC10":"0.5130","EC12":"0.4260","EC13":"0.6065","EC14":"0.2879","EC15":"1.3711","EC16":"0.5901","EC17":"0.9499","EC18":"0.7419","EC19":"0.5321","EC20":"0.7839","EC21":"0.2648","EC22":"0.9243","EC23":"1.0037","EC24":"0.7602","PVC1":"0.8079","PVC2":"0.2115","PVC3":"0.3043","PVC4":"0.3573","PVC5":"0.3705","PVC6":"0.4531","PVC7":"0.7452","PVC9":"0.4244","PVC10":"0.3800","PVC11":"0.3653","PVC12":"0.3490","PVC13":"0.4375","PVC14":"0.5620","PVC15":"0.3330","PVC16":"0.1947","PVC17":"0.3301","PVC19":"0.2837","PVC20":"0.3520","PVC21":"0.2890","PVC22":"0.9073","PVC23":"0.4906","PVC24":"0.4272"},{"Metabolite":"CE 20:1","EC1":"0.0011","EC2":"0.0029","EC3":"0.0008","EC4":"0.0034","EC5":"0.0044","EC6":"0.0104","EC7":"0.0094","EC8":"0.0021","EC10":"0.0009","EC12":"0.0021","EC13":"0.0030","EC14":"0.0034","EC15":"0.0047","EC16":"0.0033","EC17":"0.0067","EC18":"0.0045","EC19":"0.0046","EC20":"0.0017","EC21":"0.0032","EC22":"0.0035","EC23":"0.0019","EC24":"0.0061","PVC1":"0.0046","PVC2":"0.0048","PVC3":"0.0041","PVC4":"0.0036","PVC5":"0.0051","PVC6":"0.0035","PVC7":"0.0030","PVC9":"0.0010","PVC10":"0.0305","PVC11":"0.0023","PVC12":"0.0013","PVC13":"0.0020","PVC14":"0.0060","PVC15":"0.0022","PVC16":"0.0029","PVC17":"0.0017","PVC19":"0.0061","PVC20":"0.0106","PVC21":"0.0007","PVC22":"0.0038","PVC23":"0.0075","PVC24":"0.0021"},{"Metabolite":"CE 20:2","EC1":"0.0019","EC2":"0.0046","EC3":"0.0030","EC4":"0.0049","EC5":"0.0048","EC6":"0.0088","EC7":"0.0157","EC8":"0.0026","EC10":"0.0018","EC12":"0.0011","EC13":"0.0031","EC14":"0.0018","EC15":"0.0042","EC16":"0.0023","EC17":"0.0035","EC18":"0.0044","EC19":"0.0032","EC20":"0.0020","EC21":"0.0008","EC22":"0.0088","EC23":"0.0030","EC24":"0.0047","PVC1":"0.0027","PVC2":"0.0052","PVC3":"0.0024","PVC4":"0.0027","PVC5":"0.0038","PVC6":"0.0034","PVC7":"0.0005","PVC9":"0.0041","PVC10":"0.0182","PVC11":"0.0013","PVC12":"0.0020","PVC13":"0.0003","PVC14":"0.0037","PVC15":"0.0018","PVC16":"0.0058","PVC17":"0.0016","PVC19":"0.0069","PVC20":"0.0047","PVC21":"0.0003","PVC22":"0.0030","PVC23":"0.0095","PVC24":"0.0005"},{"Metabolite":"CE 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}

}