{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002858","ANALYSIS_ID":"AN004688","VERSION":"1","CREATED_ON":"September 13, 2023, 3:17 pm"},

"PROJECT":{"PROJECT_TITLE":"Intracerebroventricular Transplantation of Foetal Allogeneic Neural Stem Cells in Patients with Secondary Progressive Multiple Sclerosis (hNSC-SPMS): a phase I dose-escalation clinical trial","PROJECT_SUMMARY":"SUMMARY This is an open-label, first-in-human, dose-escalation phase I study (NCT03282760, EudraCT2015‐004855‐37) to determine the feasibility, safety, and tolerability of the transplantation of allogeneic human neural stem/progenitor cells (hNSCs) for the treatment of progressive multiple sclerosis. We report the analysis of 1 year of data from the first cohort of 15 patients from two trial sites that received increasing numbers of allogeneic hNSCs delivered via intracerebroventricular injection in combination with an immunosuppressive regimen. No treatment-related deaths nor serious adverse events (AEs) were observed over the 12-month follow-up. Participants displayed stability of clinical and laboratory parameters, as well as lesion load and activity at the brain MRIs, compared to study entry. Longitudinal metabolomics and lipidomics analyses of cerebrospinal fluid and serum from these patients identified time and dose-dependent responses, with increased levels of free fatty acids and acylcarnitines in the CSF, especially at the highest dose of injected hNSCs at the one-year follow-up time point. Finally, a significant inverse correlation was found between the highest dose of injected hNSCs and the smaller parenchymal brain volume change (PBVC; Spearman’s rho= -0.7, p= 0.03), clinical covariates that correlated with CSF levels of free fatty acids, acyl-carnitines, oxylipins, conjugated bile acids and purine breakdown and deamination products, such as hypoxanthine. The absence of AEs and the stability of functional and structural outcomes is reassuring in terms of risks and represent a main milestone to rigorously address the challenges for the safe translation of key principles of stem cell biology into effective regenerative medicines.","INSTITUTE":"University of Colorado Anschutz Medical Campus","DEPARTMENT":"Department of Biochemistry and Molecular Genetics","LAST_NAME":"Stephenson","FIRST_NAME":"Daniel","ADDRESS":"Research 1 South L18-1303 12801 E. 17th Ave., Aurora, Colorado, 80045, USA","EMAIL":"daniel.stephenson@cuanschutz.edu","PHONE":"303-724-3339"},

"STUDY":{"STUDY_TITLE":"Intracerebroventricular Transplantation of Foetal Allogeneic Neural Stem Cells in Patients with Secondary Progressive Multiple Sclerosis (hNSC-SPMS): a phase I dose-escalation clinical trial - Metabolomics Analysis of Human Serum","STUDY_SUMMARY":"This is an open-label, first-in-human, dose-escalation phase I study (NCT03282760, EudraCT2015‐004855‐37) to determine the feasibility, safety, and tolerability of the transplantation of allogeneic human neural stem/progenitor cells (hNSCs) for the treatment of progressive multiple sclerosis. We report the analysis of 1 year of data from the first cohort of 15 patients from two trial sites that received increasing numbers of allogeneic hNSCs delivered via intracerebroventricular injection in combination with an immunosuppressive regimen. No treatment-related deaths nor serious adverse events (AEs) were observed over the 12-month follow-up. Participants displayed stability of clinical and laboratory parameters, as well as lesion load and activity at the brain MRIs, compared to study entry. Longitudinal metabolomics and lipidomics analyses of cerebrospinal fluid and serum from these patients identified time and dose-dependent responses, with increased levels of free fatty acids and acylcarnitines in the CSF, especially at the highest dose of injected hNSCs at the one-year follow-up time point. Finally, a significant inverse correlation was found between the highest dose of injected hNSCs and the smaller parenchymal brain volume change (PBVC; Spearman’s rho= -0.7, p= 0.03), clinical covariates that correlated with CSF levels of free fatty acids, acyl-carnitines, oxylipins, conjugated bile acids and purine breakdown and deamination products, such as hypoxanthine. The absence of AEs and the stability of functional and structural outcomes is reassuring in terms of risks and represent a main milestone to rigorously address the challenges for the safe translation of key principles of stem cell biology into effective regenerative medicines.","INSTITUTE":"University of Colorado Anschutz Medical Campus","LAST_NAME":"Stephenson","FIRST_NAME":"Daniel","ADDRESS":"Research 1 South L18-1303 12801 E. 17th Ave., Aurora, Colorado, 80045, USA","EMAIL":"daniel.stephenson@cuanschutz.edu","PHONE":"303-724-3339"},

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{
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"Factors":{"TIMEPOINT":"REND"},
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"Subject ID":"1151",
"Sample ID":"DS1-123-020",
"Factors":{"TIMEPOINT":"FU9"},
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"Factors":{"TIMEPOINT":"REND"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-022+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-022-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-022+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-022-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-022-neg_oxylipinsSerum.mzXML"}
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"Factors":{"TIMEPOINT":"FU1"},
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"Additional sample data":{"RAW_FILE_NAME":"DS1-123-033+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-033-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-033+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-033-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-033-neg_oxylipinsSerum.mzXML"}
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},
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},
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"Additional sample data":{"RAW_FILE_NAME":"DS1-123-054+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-054-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-054+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-054-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-054-neg_oxylipinsSerum.mzXML"}
},
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"Subject ID":"1249",
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"Additional sample data":{"RAW_FILE_NAME":"DS1-123-055+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-055-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-055+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-055-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-055-neg_oxylipinsSerum.mzXML"}
},
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},
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},
{
"Subject ID":"1289",
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},
{
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},
{
"Subject ID":"1316",
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"Factors":{"TIMEPOINT":"FU9"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-062+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-062-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-062+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-062-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-062-neg_oxylipinsSerum.mzXML"}
},
{
"Subject ID":"1289",
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"Factors":{"TIMEPOINT":"FU12"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-064+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-064-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-064+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-064-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-064-neg_oxylipinsSerum.mzXML"}
},
{
"Subject ID":"1323",
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"Factors":{"TIMEPOINT":"FU6"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-065+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-065-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-065+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-065-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-065-neg_oxylipinsSerum.mzXML"}
},
{
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"Sample ID":"DS1-123-150",
"Factors":{"TIMEPOINT":"FU3"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-150+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-150-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-150+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-150-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-150-neg_oxylipinsSerum.mzXML"}
},
{
"Subject ID":"1178",
"Sample ID":"DS1-123-151",
"Factors":{"TIMEPOINT":"FU9"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-151+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-151-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-151+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-151-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-151-neg_oxylipinsSerum.mzXML"}
},
{
"Subject ID":"1219",
"Sample ID":"DS1-123-152",
"Factors":{"TIMEPOINT":"FU3"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-152+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-152-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-152+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-152-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-152-neg_oxylipinsSerum.mzXML"}
},
{
"Subject ID":"1228",
"Sample ID":"DS1-123-153",
"Factors":{"TIMEPOINT":"FU3"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-153+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-153-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-153+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-153-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-153-neg_oxylipinsSerum.mzXML"}
},
{
"Subject ID":"1219",
"Sample ID":"DS1-123-154",
"Factors":{"TIMEPOINT":"FU9"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-154+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-154-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-154+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-154-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-154-neg_oxylipinsSerum.mzXML"}
},
{
"Subject ID":"1249",
"Sample ID":"DS1-123-155",
"Factors":{"TIMEPOINT":"FU3"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-155+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-155-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-155+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-155-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-155-neg_oxylipinsSerum.mzXML"}
},
{
"Subject ID":"1228",
"Sample ID":"DS1-123-156",
"Factors":{"TIMEPOINT":"FU9"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-123-156+pos_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-156-neg_metabolomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-156+pos_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-156-neg_lipidomicsSerum.mzXML","RAW_FILE_NAME":"DS1-123-156-neg_oxylipinsSerum.mzXML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"lumbar puncture for serum and CSF collection (stored at -80°C)","SAMPLE_TYPE":"Serum"},

"TREATMENT":{"TREATMENT_SUMMARY":"A total of 180 candidates volunteered between September 26, 2017, and January 13, 2020. The study was conducted in three centers. Two Italian centers, the ‘IRCCS Casa Sollievo della Sofferenza’ Research Hospital (Site 1) and the ‘Santa Maria di Terni’ Hospital (Site 2) recruited patients and performed the MRI examinations. The Multiple Sclerosis Centre of the Neurocentre of Southern Switzerland (Site 3) performed the magnetic resonance imaging (MRI) analysis. The ICVI of the ACT was performed at Site 2. Screening was done one month prior to enrolment in the study. The run-in period started after the screening examination (Run-in-START) and lasted three months (Run-in-END). After screening in the two recruiting centers (as above), n= 15 subjects with active and non-active SPMS suffering from progressive disability were found to be eligible, consented, and were assigned a unique identification number. Participants, nine females and six males, had a mean age of 50 years (range: 38–57), and were recruited in similar proportions by the two study sites. The mean EDSS was 7.6 (range 7–8), mean disease duration was 23 years (range 14–30), and mean time from diagnosis to secondary progression was 10 years (range: 1–20). The hNSC-SPMS study was advertised using the website of the hospitals and social media (eg Facebook and Twitter). The first patient selection was performed based on the documentation provided. Selected patients were then summoned in Site 1 and Site 2 (as above) for a first pre-screening meeting with the clinicians. Patients who agreed to participate were then subjected to the screening visit for the clinical trial. Following the surgical procedure, follow-up visits were performed monthly up to 12 months. Before transplantation, all patients underwent the following examinations (at both Run-in-START and Run-in-END): physical and neurological exams; vital signs; pregnancy tests (in fertile women); haematological and urine tests; lumbar puncture for serum and CSF collection (stored at -80°C);"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Extraction of metabolites and lipids from serum and cerebrospinal fluid (CSF) was as follows: 40 µL CSF or serum was aliquoted into 2mL deep well plates followed by an addition of 360 µL cold MeOH:MeCN:H2O (5:3:2, v:v:v). Plates were then placed on a shaker at 4°C and plate shaker was set to 400 RPM for 30 minutes. Insoluble material was pelleted by centrifugation (4000 RPM, 10 min) and supernatants were isolated for analysis by UHPLC-MS. All 96-well plate pipetting was done using Integra MINI 96 (Integra Biosciences)."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Oxylipin Negative","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)","SOLVENT_A":"20:80:0.02 ACN:Water:Formic Acid","SOLVENT_B":"20:80:0.02 ACN:IPA:Formic Acid","FLOW_GRADIENT":"0min - 0.35ml/min - 0%B, 0.5min - 0.35ml/min - 25%B, 1min - 0.35ml/min - 25%B, 2.5min - 0.35ml/min - 40%B, 2.6min -0.35ml/min - 55%B, 4.5min - 0.35ml/min - 70%B, 4.6min - 0.35ml/min - 100%B, 6min - 0.35ml/min - 100%B, 6.1min - 0.35ml/min - 0%B, 7min - 0.35ml/min 0%B","FLOW_RATE":"0.35ml/min","COLUMN_TEMPERATURE":"60"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"EI","ION_MODE":"NEGATIVE","MS_COMMENTS":"Software for Feature Assignments - El-Maven"},

"MS_METABOLITE_DATA":{
"Units":"Peak area",

"Data":[{"Metabolite":"12(13)-DiHOME","DS1-123-002":"53094.96","DS1-123-004":"82547.93","DS1-123-006":"174836.83","DS1-123-008":"92728.73","DS1-123-009":"111575.84","DS1-123-010":"80008.46","DS1-123-012":"125325.5","DS1-123-013":"75116.18","DS1-123-016":"129186.29","DS1-123-017":"143107.98","DS1-123-019":"66350.7","DS1-123-020":"97663.85","DS1-123-022":"200561.75","DS1-123-023":"88842.22","DS1-123-024":"87265.79","DS1-123-026":"98172.75","DS1-123-029":"261680.44","DS1-123-030":"283628.12","DS1-123-033":"98368.81","DS1-123-034":"143824.31","DS1-123-036":"225608.33","DS1-123-038":"165948.7","DS1-123-040":"128943.5","DS1-123-041":"121971.56","DS1-123-044":"100796.59","DS1-123-045":"22640.62","DS1-123-047":"172166.77","DS1-123-048":"119817.87","DS1-123-049":"95327.46","DS1-123-052":"70576.51","DS1-123-053":"216411.5","DS1-123-054":"145468.91","DS1-123-055":"358574","DS1-123-056":"181613.5","DS1-123-059":"82280.09","DS1-123-060":"99776.53","DS1-123-061":"83063.16","DS1-123-062":"187147.92","DS1-123-064":"146084.62","DS1-123-065":"82840.66","DS1-123-066":"97784.23","DS1-123-067":"66168.15","DS1-123-068":"78910.27","DS1-123-069":"83798.89","DS1-123-074":"75563.66","DS1-123-075":"166851.23","DS1-123-077":"91343.97","DS1-123-078":"94450.55","DS1-123-079":"126252.76","DS1-123-082":"100100.93","DS1-123-084":"183046.17","DS1-123-086":"62821","DS1-123-088":"142940.02","DS1-123-089":"81924.55","DS1-123-090":"109552.12","DS1-123-091":"117368.32","DS1-123-092":"257667.45","DS1-123-095":"300018.5","DS1-123-096":"66638.82","DS1-123-097":"115763.88","DS1-123-098":"112486.23","DS1-123-101":"95793.7","DS1-123-104":"157763.48","DS1-123-107":"128334.63","DS1-123-108":"106178.12","DS1-123-110":"112791.23","DS1-123-112":"78414.47","DS1-123-113":"95551.44","DS1-123-115":"85448.76","DS1-123-118":"197749.73","DS1-123-119":"97618.47","DS1-123-120":"112803.84","DS1-123-121":"113231.87","DS1-123-122":"91945.94","DS1-123-126":"220498.8","DS1-123-128":"149488.25","DS1-123-129":"216096.38","DS1-123-132":"112775.53","DS1-123-135":"88643.66","DS1-123-137":"127678.98","DS1-123-138":"189915.11","DS1-123-140":"71850.12","DS1-123-141":"135191.16","DS1-123-143":"201770.73","DS1-123-144":"86207.78","DS1-123-147":"85290.15","DS1-123-149":"116607.48","DS1-123-150":"199075.3","DS1-123-151":"94331.13","DS1-123-152":"111108.45","DS1-123-153":"100699.23","DS1-123-154":"109320.73","DS1-123-155":"100339.73","DS1-123-156":"85828.54"},{"Metabolite":"12(S)-HETE","DS1-123-002":"1532997.12","DS1-123-004":"293299.84","DS1-123-006":"318151","DS1-123-008":"1572566.38","DS1-123-009":"156221.69","DS1-123-010":"2132392","DS1-123-012":"299193.53","DS1-123-013":"2136367","DS1-123-016":"1500540.62","DS1-123-017":"214908.86","DS1-123-019":"963360.69","DS1-123-020":"105983.09","DS1-123-022":"172175.41","DS1-123-023":"1393806.88","DS1-123-024":"101052.78","DS1-123-026":"586033.25","DS1-123-029":"135815.42","DS1-123-030":"94877.82","DS1-123-033":"668352.31","DS1-123-034":"45495.56","DS1-123-036":"229974.75","DS1-123-038":"170959.3","DS1-123-040":"1373084.38","DS1-123-041":"197775.58","DS1-123-044":"48423.97","DS1-123-045":"0","DS1-123-047":"61309.98","DS1-123-048":"51947.45","DS1-123-049":"18816.38","DS1-123-052":"855395.44","DS1-123-053":"174083.05","DS1-123-054":"83837.21","DS1-123-055":"23810.47","DS1-123-056":"536677","DS1-123-059":"83654.14","DS1-123-060":"485728","DS1-123-061":"1418991","DS1-123-062":"144918.98","DS1-123-064":"419757.28","DS1-123-065":"109407.68","DS1-123-066":"287904.41","DS1-123-067":"1061857.38","DS1-123-068":"852474.31","DS1-123-069":"2733160.75","DS1-123-074":"2689071.75","DS1-123-075":"44956.45","DS1-123-077":"952477.31","DS1-123-078":"246527.5","DS1-123-079":"79611.69","DS1-123-082":"3140816.25","DS1-123-084":"760945.69","DS1-123-086":"6269872","DS1-123-088":"1180802.38","DS1-123-089":"615202.88","DS1-123-090":"154488.89","DS1-123-091":"2034304","DS1-123-092":"678831.25","DS1-123-095":"2509443","DS1-123-096":"2223902.25","DS1-123-097":"4647949","DS1-123-098":"2058190","DS1-123-101":"1478165","DS1-123-104":"2078787.62","DS1-123-107":"35163.22","DS1-123-108":"584812.81","DS1-123-110":"2497004","DS1-123-112":"1451707.62","DS1-123-113":"1512516","DS1-123-115":"3128231.75","DS1-123-118":"1389738.5","DS1-123-119":"364008.25","DS1-123-120":"1987669","DS1-123-121":"199688.45","DS1-123-122":"1754114","DS1-123-126":"215535.75","DS1-123-128":"1008013.75","DS1-123-129":"239210.45","DS1-123-132":"658983.69","DS1-123-135":"10495953","DS1-123-137":"10110801","DS1-123-138":"1328819.12","DS1-123-140":"671078.06","DS1-123-141":"2314841.25","DS1-123-143":"451376.84","DS1-123-144":"995266.19","DS1-123-147":"2378393.5","DS1-123-149":"2798901","DS1-123-150":"205257.3","DS1-123-151":"207177.31","DS1-123-152":"1055414.12","DS1-123-153":"120117.05","DS1-123-154":"1574524","DS1-123-155":"52905.68","DS1-123-156":"393071.25"},{"Metabolite":"13(S)-HODE","DS1-123-002":"94498.69","DS1-123-004":"167712.25","DS1-123-006":"308153.69","DS1-123-008":"251623.56","DS1-123-009":"284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}

}