{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001660","ANALYSIS_ID":"AN002711","VERSION":"1","CREATED_ON":"January 25, 2021, 3:47 pm"},

"PROJECT":{"PROJECT_TITLE":"Plasmodium falciparum metabolomics as a result of treatment with putative acetyl-CoA synthetase inhibitors","PROJECT_SUMMARY":"Plasmodium falciparum is the most virulent species of parasites that cause malaria. Current drug efforts that are used to combat this deadly disease often employ pharmacologic strategies that inhibit critical parasite metabolic functions. Compounds used in the present study have generated resistance mutations in major acetyl-CoA producing enzymes. Hence, we set out to test the metabolic effects of these compounds on Plasmodium falciparum parasites including those on acetyl-CoA production by treating parasites for 2.5 hours under 10xIC50 of the test compounds. We used a targeted HPLC/mass spectrometry-based approach to analyze parasite metabolism. We find that multiple compounds tested in these studies have led to relative decreases in acetyl-CoA abundance compared to control parasite conditions. These studies are necessary for understanding pharmacology effects on the most virulent human malaria parasite.","INSTITUTE":"Pennsylvania State University","DEPARTMENT":"Chemistry","LABORATORY":"Llinás Laboratory","LAST_NAME":"Llinás","FIRST_NAME":"Manuel","ADDRESS":"W126 Millennium Science Complex, University Park, PENNSYLVANIA, 16802, USA","EMAIL":"mul27@psu.edu","PHONE":"814-867-3444","FUNDING_SOURCE":"This work was supported by the Bill and Melinda Gates Foundation (OPP1054480)"},

"STUDY":{"STUDY_TITLE":"Plasmodium falciparum metabolomics as a result of treatment with putative acetyl-CoA synthetase inhibitors","STUDY_SUMMARY":"Plasmodium falciparum cells in culture were treated with respective compounds for 2.5 hours at 10xIC50 values. Metabolites were isolated using 90% methanol, dried, reconstituted in HPLC-grade water, and analyzed by HPLC/MS. Resulting data were analyzed and compiled to generate study data.","INSTITUTE":"Pennsylvania State University","DEPARTMENT":"Chemistry","LABORATORY":"Llinás Laboratory","LAST_NAME":"Llinás","FIRST_NAME":"Manuel","ADDRESS":"W126 Millennium Science Complex, University Park, PENNSYLVANIA, 16802, USA","EMAIL":"mul27@psu.edu","PHONE":"814-867-3444","NUM_GROUPS":"3","TOTAL_SUBJECTS":"24"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Plasmodium falciparum","TAXONOMY_ID":"5833","GENOTYPE_STRAIN":"3D7","GENDER":"Not applicable","CELL_COUNTS":"1x10^8"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"20171106-MMV084978-1",
"Factors":{"Treatment":"MMV084978"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-MMV084978-1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-MMV084978-2",
"Factors":{"Treatment":"MMV084978"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-MMV084978-2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-MMV084978-3",
"Factors":{"Treatment":"MMV084978"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-MMV084978-3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-Blank1",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-Blank1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-Blank2",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-Blank2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-Blank3",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-Blank3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-ND1",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-ND1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-ND2",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-ND2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-ND3",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-ND3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-Pool1",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-Pool1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-Pool2",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-Pool2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20171106-Pool3",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20171106-Pool3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-MMV019721-1",
"Factors":{"Treatment":"MMV019721"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-MMV019721-1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-MMV019721-2",
"Factors":{"Treatment":"MMV019721"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-MMV019721-2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-MMV019721-3",
"Factors":{"Treatment":"MMV019721"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-MMV019721-3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-Blank1",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-Blank1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-Blank2",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-Blank2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-Blank3",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-Blank3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-Blank4",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-Blank4.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-Blank5",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-Blank5.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-ND1",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-ND1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-ND2",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-ND2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-ND3",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-ND3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-QC1",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-QC1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-QC2",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-QC2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20180815-QC3",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20180815-QC3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-MMV019721-1",
"Factors":{"Treatment":"MMV019721"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-MMV019721-1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-MMV019721-2",
"Factors":{"Treatment":"MMV019721"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-MMV019721-2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-MMV019721-3",
"Factors":{"Treatment":"MMV019721"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-MMV019721-3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-Blank1",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-Blank1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-Blank2",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-Blank2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-Blank3",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-Blank3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-Blank4",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-Blank4.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-Blank5",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-Blank5.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-ND1",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-ND1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-ND2",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-ND2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-ND3",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-ND3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-QC1",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-QC1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-QC2",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-QC2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181008-QC3",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20181008-QC3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-MMV084978-1",
"Factors":{"Treatment":"MMV084978"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-MMV084978-1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-MMV084978-2",
"Factors":{"Treatment":"MMV084978"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-MMV084978-2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-MMV084978-3",
"Factors":{"Treatment":"MMV084978"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-MMV084978-3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-Blank1",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-Blank1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-Blank2",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-Blank2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-Blank3",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-Blank3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-Blank4",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-Blank4.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-Blank5",
"Factors":{"Treatment":"Blank"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-Blank5.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-ND1",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-ND1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-ND2",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-ND2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-ND3",
"Factors":{"Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-ND3.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-QC1",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-QC1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-QC2",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-QC2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"20181220-QC3",
"Factors":{"Treatment":"Pooled Parasite Samples"},
"Additional sample data":{"RAW_FILE_NAME":"20181220-QC3.mzXML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Plasmodium falciparum 3D7 parasites were cultured in RPMI 1640 medium and magnetically enriched to increase the infected to uninfected RBC ratio. Following hemocytometer counts, 1x10^8 parasites were measured per condition into 5 mL of total media for 2.5 hours in the presence of drug. Medium was aspirated until 1 mL remained on each sample, the sample was transferred to a micro-centrifuge tube, spun, and the medium was aspirated.","COLLECTION_PROTOCOL_FILENAME":"Metabolite_Extraction_for_LCMS_2017.pdf","SAMPLE_TYPE":"Cultured cells","COLLECTION_LOCATION":"Millenium Science Complex, University Park, Pennsylvania","STORAGE_CONDITIONS":"-80℃","COLLECTION_VIALS":"1.5 mL eppendorf","STORAGE_VIALS":"1.5 mL eppendorf","COLLECTION_TUBE_TEMP":"On ice","TISSUE_CELL_QUANTITY_TAKEN":"1x10^8 cells per sample in 1 mL total volume"},

"TREATMENT":{"TREATMENT_SUMMARY":"Plasmodium falciparum 3D7 parasites were cultured in RPMI 1640 medium and magnetically enriched to increase the infected to uninfected RBC ratio. Following hemocytometer counts, 1x10^8 parasites were measured per condition into 5 mL of total media for 2.5 hours in the presence of drug. No treatment or ND represents control parasites without additional alterations. Compounds (MMV...) are test compounds that were added at 10xIC50 value for the experimental duration. Blanks are sample tubes that follow the same procedures as samples following the quenching of metabolism. Both Pool and QC samples are combined samples of all samples from the analytical batch on that particular day. Dates in YYYYMMDD format are appended to individual samples to indicate the batch in which they were processed.","TREATMENT_PROTOCOL_FILENAME":"Metabolite_Extraction_for_LCMS_2017.pdf","TREATMENT_ROUTE":"Transfer to media by pipette","TREATMENT_DOSE":"10xIC50","TREATMENT_DOSEVOLUME":"5 uL or less","TREATMENT_DOSEDURATION":"2.5 hours","TREATMENT_VEHICLE":"DMSO","CELL_STORAGE":"Temperature and gas composition controlled incubator","CELL_GROWTH_CONTAINER":"6-well plate","CELL_GROWTH_CONFIG":"5 mL in each sample well, 1x10^8 infected red blood cells per well, conditions performed in triplicate","CELL_MEDIA":"RPMI 1640 containing Albumax, Gentamycin, Hypoxanthine, HEPES, Sodium Bicarbonate","TREATMENT_COMPOUND":"MMV084978, MMV019721"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Once samples were obtained, metabolism was quenched with 90% methanol containing 0.25 uM labeled aspartate, cells were centrifuged, and the supernatant was removed by nitrogen drying. Sample was reconstituted in 3% HPLC-grade methanol and run on the instrument.","PROCESSING_METHOD":"Wash, spin, quench, spin, dry, store at -80 C until run on the instrument, resuspend","PROCESSING_STORAGE_CONDITIONS":"On ice","EXTRACTION_METHOD":"90% methanol extraction","EXTRACT_CLEANUP":"Centrifugation and nitrogen drying","SAMPLE_RESUSPENSION":"100 uL 3% methanol with 1 uM chlorpropamide","SAMPLE_SPIKING":"0.25 uM Labeled Aspartate, 1 uM chlorpropamide"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Ion-pairing method using reverse-phase chromatography setup.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Waters XSelect HSS (2.1 x 100 mm, 2.5 um)","COLUMN_TEMPERATURE":"30","SOLVENT_A":"3% methanol, 15 mM acetic acid, 10 mM tributylamine","SOLVENT_B":"100% methanol"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Plus Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"Data was centroided using MSConvert and converted to .mzXML for utilization in MzMine and El-Maven software."},

"MS_METABOLITE_DATA":{
"Units":"Normalized and blank subtracted peak area",

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}

}