{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002432","ANALYSIS_ID":"AN003960","VERSION":"1","CREATED_ON":"January 5, 2023, 10:27 am"},

"PROJECT":{"PROJECT_TITLE":"Metabolic impacts of metformin to seasonal influenza vaccination: a pilot study of drug interaction with the immune response","PROJECT_SUMMARY":"We report here a double-blinded pilot study of seasonal influenza vaccination, where half of the participants received daily metformin administration. Global metabolomics was measured in the plasma samples at six timepoints. Metformin signatures were successfully identified in the metabolomics data. Statistically significant metabolite features were found both for the vaccination effect and for the drug-vaccine interactions.","INSTITUTE":"The Jackson Laboratory For Genomic Medicine","LABORATORY":"Shuzhao Li Lab","LAST_NAME":"Siddiqa","FIRST_NAME":"Amnah","ADDRESS":"10 Discovery Drive , Farmington, CT 06032","EMAIL":"amnah.siddiqa@jax.org","PHONE":"4049187223"},

"STUDY":{"STUDY_TITLE":"Metabolic impacts of metformin to seasonal influenza vaccination: a pilot study of drug interaction with the immune response","STUDY_SUMMARY":"We report here a double-blinded pilot study of seasonal influenza vaccination, where half of the participants received daily metformin administration. Global metabolomics was measured in the plasma samples at six timepoints. Metformin signatures were successfully identified in the metabolomics data. Statistically significant metabolite features were found both for the vaccination effect and for the drug-vaccine interactions.","INSTITUTE":"The Jackson Laboratory For Genomic Medicine","LABORATORY":"Shuzhao Li Lab","LAST_NAME":"Siddiqa","FIRST_NAME":"Amnah","ADDRESS":"10 Discovery dr, Farmington, CT 06032","EMAIL":"amnah.siddiqa@jax.org","PHONE":"4049187223"},

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{
"Subject ID":"-",
"Sample ID":"YW_20201210_234.mzML",
"Factors":{"Visit":"V6","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_234.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_102.mzML",
"Factors":{"Visit":"V1","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_102.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_094.mzML",
"Factors":{"Visit":"V2","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_094.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_098.mzML",
"Factors":{"Visit":"V3","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_098.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_104.mzML",
"Factors":{"Visit":"V4","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_104.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_096.mzML",
"Factors":{"Visit":"V5","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_096.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_100.mzML",
"Factors":{"Visit":"V6","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_100.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_200.mzML",
"Factors":{"Visit":"V1","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_200.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_208.mzML",
"Factors":{"Visit":"V2","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_208.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_210.mzML",
"Factors":{"Visit":"V3","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_210.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_202.mzML",
"Factors":{"Visit":"V4","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_202.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_204.mzML",
"Factors":{"Visit":"V5","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_204.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_206.mzML",
"Factors":{"Visit":"V6","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_206.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_228.mzML",
"Factors":{"Visit":"V1","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_228.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_220.mzML",
"Factors":{"Visit":"V2","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_220.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_218.mzML",
"Factors":{"Visit":"V3","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_218.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_224.mzML",
"Factors":{"Visit":"V4","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_224.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_222.mzML",
"Factors":{"Visit":"V5","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_222.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_226.mzML",
"Factors":{"Visit":"V6","Treatment":"Metformin"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_226.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_058.mzML",
"Factors":{"Visit":"V1","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_058.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_056.mzML",
"Factors":{"Visit":"V2","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_056.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_052.mzML",
"Factors":{"Visit":"V3","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_052.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_054.mzML",
"Factors":{"Visit":"V4","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_054.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_050.mzML",
"Factors":{"Visit":"V5","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_050.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_048.mzML",
"Factors":{"Visit":"V6","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_048.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_246.mzML",
"Factors":{"Visit":"V1","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_246.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_248.mzML",
"Factors":{"Visit":"V2","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_248.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_254.mzML",
"Factors":{"Visit":"V3","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_254.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_252.mzML",
"Factors":{"Visit":"V4","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_252.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_250.mzML",
"Factors":{"Visit":"V5","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_250.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_256.mzML",
"Factors":{"Visit":"V6","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_256.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_194.mzML",
"Factors":{"Visit":"V1","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_194.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_192.mzML",
"Factors":{"Visit":"V2","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_192.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_188.mzML",
"Factors":{"Visit":"V3","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_188.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_196.mzML",
"Factors":{"Visit":"V4","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_196.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_190.mzML",
"Factors":{"Visit":"V5","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_190.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_186.mzML",
"Factors":{"Visit":"V6","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_186.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_140.mzML",
"Factors":{"Visit":"V1","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_140.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_142.mzML",
"Factors":{"Visit":"V2","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_142.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_146.mzML",
"Factors":{"Visit":"V3","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_146.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_150.mzML",
"Factors":{"Visit":"V4","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_150.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_144.mzML",
"Factors":{"Visit":"V5","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_144.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"YW_20201210_148.mzML",
"Factors":{"Visit":"V6","Treatment":"Placebo"},
"Additional sample data":{"RAW_FILE_NAME":"YW_20201210_148.mzML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"EDTA-treated whole blood was immediately centrifuged and the resultant plasma was stored at -80˚C until analyses. Plasma metabolites extraction was carried out by protein precipitation technique using extraction solvent, acetonitrile:methanol (8:1, v/v) containing 0.1% formic acid. All samples were vortexed and incubated with shaking at 1000 rpm for 10 min at 4˚C followed by centrifugation at 4˚ C for 15 min at 15,000 rpm. The supernatant was transferred into mass spec vials and 2 micro liter injected into UHPLC-MS.","SAMPLE_TYPE":"Blood (plasma)","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"This pilot study is a double-blinded placebo-controlled trial in men and women over the age of 65 years. Subjects were screened rigorously for eligibility. Study exclusion criteria included the following: any unstable medical conditions or severe co-morbidities (severe COPD, severe congestive heart failure, advanced neurological disorders, etc), contraindications for metformin (severe renal or liver impairment), contraindication for flu vaccine (history of Guillain-Barre syndrome post vaccination or allergic to component of vaccine), immunosuppressive disorders, immunosuppressive medications, and active cancer. Importantly, participants were excluded if they were prediabetic or diabetic (HbA1c ≥ 5.7%) to avoid any confounding impact of metformin on diabetes status. Eligible participants were randomized to metformin (final dose 1500mg extended release (ER)/day) or placebo treatment. To limit gastrointestinal issues per current metformin label recommendations, participants started with 1 tablet a day for week 1 (500mg metformin ER/day or placebo), then 2 tablets a day for week 2 (1000mg metformin ER or placebo), and finally 3 tablets a day for week 3 until the completion of the study (1500mg metformin ER or placebo). Fifteen subjects (n=8 metformin, n=7 placebo) were randomized and completed the study on treatment with no differences in basic characteristics at baseline (placebo: 74.71±2.45 years old, 3 males, BMI: 27.31±1.68; metformin: 74.13±2.42 years old, 5 males, BMI: 26.43±1.47). All participants were vaccinated with Fluzone high-dose trivalent flu vaccine (Sanofi Pasteur Inc., Swiftwater, PA) after ~70 days of treatment. Blood was drawn via standard venipuncture into EDTA-treated vacutainers prior to treatment (Day 0), prior to vaccination (~day 35 and ~day 70), and 7, ~35, and ~70 days post vaccination. The study protocol was approved by the Institutional Review Board at the University of Connecticut Health Center (UCHC) and registered at ClinicalTrials.gov (NCT03996538). All study participants provided written informed consent to participate in the study."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"The plasma samples were thawed in ice. Metabolites extraction were carried out by protein precipitation using extraction solvent, acetonitrile:methanol (8:1, v/v) containing 0.1% formic acid and isotope labelled Trimethyl-13C3]-caffeine, [13C5]-L-glutamic acid, [15N2]-Uracil, [15N,13C5]-L-methionine, [13C6]-D-glucose and [15N]-L-tyrosin as spike-in controls. 30 μl of plasma was taken and 60 μl of extraction solvent was added. All samples were vortexed and incubated with shaking at 1000 rpm for 10 min at 4°C followed by centrifugation at 4°C for 15 min at 15,000 rpm. The supernatant was transferred into mass spec vials and 2 μl injected into UHPLC-MS."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Thermo ScientificTM TranscendTM Duo LX-2 UHPLC","COLUMN_NAME":"HILIC column, AccucoreTM-150-Amide (50 x 2.1mm,2.6um)","SOLVENT_A":"95% acetonitrile/5% water; 0.1% acetic acid; 10 mM ammonium acetate","SOLVENT_B":"50% acetonitrile/50% water; 0.1% acetic acid; 10 mM ammonium acetate","FLOW_GRADIENT":"0.55 ml/min: 0-0.1 min: 0% B, 0.10-5.0 min: 98% B, and 5 min for cleaning and equilibration of column.","FLOW_RATE":"0.55 ml/min","COLUMN_TEMPERATURE":"45"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Orbitrap ID-X tribrid","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"Asari was used for data processing","MS_RESULTS_FILE":"ST002432_AN003960_Results.txt UNITS:m/z Has m/z:Yes Has RT:Yes RT units:Seconds"}

}