{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000847","ANALYSIS_ID":"AN001371","VERSION":"1","CREATED_ON":"August 10, 2017, 3:01 pm"},

"PROJECT":{"PROJECT_TITLE":"Statin Immuno-Metabolomics in Asthma","PROJECT_TYPE":"Placebo-controled trial","PROJECT_SUMMARY":"Innovative and novel therapies are urgently needed for the treatment of patients with severe asthma, especially those who are refractory to standard-of-care bronchodilators and inhaled corticosteroids. The Zeki lab is investigating the role of the mevalonate (MA) pathway, in the pathogenesis of airway inflammation and remodeling. Although statins all inhibit HMGCR in the same manner in terms of enzyme binding site, the statins’ varied physiochemical properties with respect to their polarity (i.e. lipophilicity) result in very different immune and lipid effects. The major significance of this work is to advance a new class of inhaler therapies for asthma; the statins which work by an entirely different mechanism than current ICS/LABA mainstays. Evidence suggests that statins may have an additive benefit to corticosteroids in asthma, thereby confirming a unique mechanism, namely via MVA pathway inhibition. This becomes particularly important in the severe asthma population which is highly corticosteroid-resistant, is poorly controlled with high exacerbation rates and hospitalizations, and has the highest healthcare costs of all asthma phenotypes. In essence, the potential public health impact of even an incremental improvement in asthma symptom control cannot be underestimated. Even the prevention of 1 asthma attack preserves lung function and reduces the adverse personal and financial impact. This study aimed to determine if statin polarity affects airway drug concentration and systemic drug absorption and to determine the effect of inhaled statins on naïve airway immune cell populations and alveolar-capillary membrane and epithelial barrier integrity in healthy rhesus monkeys. In this particular component of the study, we investigated the metabolic effects resulting from the use of statins in these healthy rhesus monkeys. Specifically, the Newman lab analyzed for lipid mediator (oxylipin, endocannabinoid, fatty acid, and nitro lipid) in lung and trachea tissue, plasma, and BAL and bile acid changes in the lung and trachea tissue and plasma.","INSTITUTE":"University of California, Davis","DEPARTMENT":"Internal Medicine","LAST_NAME":"Zeki","FIRST_NAME":"Amir","ADDRESS":"2825 J St. Suite 400 Sacramento, CA 95816","EMAIL":"aazeki@ucdavis.edu","PHONE":"(916) 734-8230"},

"STUDY":{"STUDY_TITLE":"Statin Immuno-Metabolomics in Asthma (part V)","STUDY_TYPE":"Placebo-controled trial","STUDY_SUMMARY":"Innovative and novel therapies are urgently needed for the treatment of patients with severe asthma, especially those who are refractory to standard-of-care bronchodilators and inhaled corticosteroids. The Zeki lab is investigating the role of the mevalonate (MA) pathway, in the pathogenesis of airway inflammation and remodeling. Although statins all inhibit HMGCR in the same manner in terms of enzyme binding site, the statins’ varied physiochemical properties with respect to their polarity (i.e. lipophilicity) result in very different immune and lipid effects. The major significance of this work is to advance a new class of inhaler therapies for asthma; the statins which work by an entirely different mechanism than current ICS/LABA mainstays. Evidence suggests that statins may have an additive benefit to corticosteroids in asthma, thereby confirming a unique mechanism, namely via MVA pathway inhibition. This becomes particularly important in the severe asthma population which is highly corticosteroid-resistant, is poorly controlled with high exacerbation rates and hospitalizations, and has the highest healthcare costs of all asthma phenotypes. In essence, the potential public health impact of even an incremental improvement in asthma symptom control cannot be underestimated. Even the prevention of 1 asthma attack preserves lung function and reduces the adverse personal and financial impact. This study aimed to determine if statin polarity affects airway drug concentration and systemic drug absorption and to determine the effect of inhaled statins on naïve airway immune cell populations and alveolar-capillary membrane and epithelial barrier integrity in healthy rhesus monkeys. In this particular component of the study, we investigated the metabolic effects resulting from the use of statins in these healthy rhesus monkeys. Specifically, the Newman lab analyzed for lipid mediator (oxylipin, endocannabinoid, fatty acid, and nitro lipid) in lung and trachea tissue, plasma, and BAL and bile acid changes in the lung and trachea tissue and plasma.","INSTITUTE":"USDA","DEPARTMENT":"Obesity and metabolism research unit","LAST_NAME":"Newman","FIRST_NAME":"John","ADDRESS":"430 West Health Sciences Dr. Davis, Ca, 95616","EMAIL":"John.Newman@ars.usda.gov","PHONE":"(530) 752-1009"},

"SUBJECT":{"SUBJECT_TYPE":"Animal","SUBJECT_SPECIES":"Macaca mulatta","TAXONOMY_ID":"9544"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Zeki R 74",
"Factors":{"Tissue":"Right Lung","Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki R 28 Repl Avg.",
"Factors":{"Tissue":"Right Lung","Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki R 48",
"Factors":{"Tissue":"Right Lung","Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki R 68",
"Factors":{"Tissue":"Right Lung","Treatment":"Simvastatin"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki R 40",
"Factors":{"Tissue":"Right Lung","Treatment":"Simvastatin"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki R 95",
"Factors":{"Tissue":"Right Lung","Treatment":"Simvastatin"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki L 74",
"Factors":{"Tissue":"Left Lung","Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki L 48",
"Factors":{"Tissue":"Left Lung","Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki L 28 Repl Avg.",
"Factors":{"Tissue":"Left Lung","Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki L 95",
"Factors":{"Tissue":"Left Lung","Treatment":"Simvastatin"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki L 40",
"Factors":{"Tissue":"Left Lung","Treatment":"Simvastatin"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki L 68",
"Factors":{"Tissue":"Left Lung","Treatment":"Simvastatin"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki T 74",
"Factors":{"Tissue":"Trachea","Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki T 48",
"Factors":{"Tissue":"Trachea","Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki T 28 Repl Avg.",
"Factors":{"Tissue":"Trachea","Treatment":"Control"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki T 40",
"Factors":{"Tissue":"Trachea","Treatment":"Simvastatin"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki T 68",
"Factors":{"Tissue":"Trachea","Treatment":"Simvastatin"}
},
{
"Subject ID":"-",
"Sample ID":"Zeki T 95",
"Factors":{"Tissue":"Trachea","Treatment":"Simvastatin"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Monkeys were treated with placebo or Provastatin for 12 days. Further, after the wash out period animals were treated with Simvastatin for 12 days. Plasma was collected at day 0, 8 and 12 of each treatment.","SAMPLE_TYPE":"Tissue","TISSUE_CELL_IDENTIFICATION":"Trachea and lung tissue"},

"TREATMENT":{"TREATMENT_SUMMARY":"Monkeys were treated (by inhalation) with placebo or Provastatin for 12 days. Further, after the wash out period animals were treated with Simvastatin for 12 days. Plasma was collected at day 0, 8 and 12 of each treatment."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Oxylipins, endocannabinoids, bile acids and fatty acids were isolated using a Waters Ostro Sample Preparation Plate (Milford, MA). Lung and trachea samples were aliquoted (~40-60mg) into 2mL polypropylene tubes and spiked with a 5 µL anti-oxidant solution (0.2 mg/ml solution BHT/EDTA in 1:1 MeOH:water) and 10 μL 1000nM analytical deuterated surrogates. A total of 50 µL of methanol, 550µL isopropanol w/ 10mM ammonium formate & 1% formic acid and 100 uL water were added and the tube was placed in a Geno/Grinder for 30 sec before being centrifuged at 10,000g for 5 min at room temp. The supernate was then transferred into the plate wells and samples were eluted into glass inserts containing 10 μL 20% glycerol by applying a vacuum at 15 Hg for 10 min. Eluent was dried by speed vacuum for 35 min at the medium BP setting, before switching to an aqueous setting for an additional 35 min. Once dry, samples were re-constituted with the internal standard 1-cyclohexyl ureido, 3-dodecanoic acid (CUDA) and 1-Phenyl 3-Hexadecanoic Acid Urea (PHAU) at 100 nM (50:50 MeOH:CAN), vortexed 1 min, transferred to a spin filter (0.1 µm, Millipore, Billerica, MA), centrifuged for 3 min at 6ºC at <4500g (rcf), before being transferred to 2 mL LC-MS amber vials. Extracts were stored at -20ºC until analysis by UPLC-MS/MS. The internal standard was used to quantify the recovery of surrogate standards."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Waters Acquity","COLUMN_NAME":"Aquity C18 BEH 1.7µm 100mm x 2.1mm column","FLOW_GRADIENT":"See protocol/methods file","FLOW_RATE":"0.4 mL/min","COLUMN_TEMPERATURE":"60 °C","SOLVENT_A":"0.1% Formic Acid","SOLVENT_B":"0.1% Formic Acid in Acetonitrile","INTERNAL_STANDARD":"See protocol/methods file","RETENTION_TIME":"See protocol/methods file","SAMPLE_INJECTION":"5 µL","ANALYTICAL_TIME":"16 min","WEAK_WASH_SOLVENT_NAME":"20% methanol, 10% isopropanol","WEAK_WASH_VOLUME":"600 µL","STRONG_WASH_SOLVENT_NAME":"50:50 Acetonitrile:Methanol","STRONG_WASH_VOLUME":"600 µL"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"Newman","OPERATOR_NAME":"Kamil Borkowski","DETECTOR_TYPE":"API 6500 QTrap (AB Sciex, Framingham, MA, USA)","SOFTWARE_VERSION":"AB Sciex MultiQuant version 3.0.2","DATA_FORMAT":".mzML"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"ABI Sciex 6500 QTrap","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"NEGATIVE"},

"MS_METABOLITE_DATA":{
"Units":"Concentration (nM)",

"Data":[{"Metabolite":"CA","Zeki R 74":"11.3","Zeki R 28 Repl Avg.":"10.55","Zeki R 48":"12.5","Zeki R 68":"35.6","Zeki R 40":"10.8","Zeki R 95":"468","Zeki L 74":"423","Zeki L 48":"6.58","Zeki L 28 Repl Avg.":"34.1","Zeki L 95":"16","Zeki L 40":"31.5","Zeki L 68":"27.3","Zeki T 74":"46.9","Zeki T 48":"21.1","Zeki T 28 Repl Avg.":"35.3","Zeki T 40":"86","Zeki T 68":"14.8","Zeki T 95":"455"},{"Metabolite":"CDCA","Zeki R 74":"3.42","Zeki R 28 Repl Avg.":"8.87","Zeki R 48":"12.3","Zeki R 68":"118","Zeki R 40":"4.9","Zeki R 95":"494","Zeki L 74":"441","Zeki L 48":"9.16","Zeki L 28 Repl Avg.":"64.5","Zeki L 95":"21.5","Zeki L 40":"77.7","Zeki L 68":"19.8","Zeki T 74":"398","Zeki T 48":"144","Zeki T 28 Repl Avg.":"302","Zeki T 40":"69.7","Zeki T 68":"8.3","Zeki T 95":"453"},{"Metabolite":"TCA","Zeki R 74":"58.9","Zeki R 28 Repl Avg.":"19.14","Zeki R 48":"96","Zeki R 68":"17","Zeki R 40":"49.9","Zeki R 95":"13.5","Zeki L 74":"11.6","Zeki L 48":"39.6","Zeki L 28 Repl Avg.":"16.3","Zeki L 95":"5.5","Zeki L 40":"33.1","Zeki L 68":"14.1","Zeki T 74":"171","Zeki T 48":"249","Zeki T 28 Repl Avg.":"5.47","Zeki T 40":"272","Zeki T 68":"23.2","Zeki T 95":"12.6"},{"Metabolite":"TCDCA","Zeki R 74":"66","Zeki R 28 Repl Avg.":"69.34","Zeki R 48":"64.8","Zeki R 68":"60.2","Zeki R 40":"58.5","Zeki R 95":"52.4","Zeki L 74":"49.9","Zeki L 48":"80.7","Zeki L 28 Repl Avg.":"63.75","Zeki L 95":"22.7","Zeki L 40":"99.5","Zeki L 68":"62.8","Zeki T 74":"279","Zeki T 48":"1160","Zeki T 28 Repl Avg.":"53.2","Zeki T 40":"409","Zeki T 68":"139","Zeki T 95":"45.4"},{"Metabolite":"GCA","Zeki R 74":"213","Zeki R 28 Repl Avg.":"92.6","Zeki R 48":"200","Zeki R 68":"106","Zeki R 40":"200","Zeki R 95":"89.9","Zeki L 74":"80.1","Zeki L 48":"163","Zeki L 28 Repl Avg.":"174","Zeki L 95":"26.3","Zeki L 40":"257","Zeki L 68":"202","Zeki T 74":"618","Zeki T 48":"399","Zeki T 28 Repl Avg.":"92.6","Zeki T 40":"2110","Zeki T 68":"94.3","Zeki T 95":"80.4"},{"Metabolite":"GCDCA","Zeki R 74":"568","Zeki R 28 Repl 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48":"5.53","Zeki T 28 Repl Avg.":"4.8","Zeki T 40":"5.54","Zeki T 68":"3.31","Zeki T 95":"7.59"},{"Metabolite":"TDCA","Zeki R 74":"35.9","Zeki R 28 Repl Avg.":"21.28","Zeki R 48":"34.7","Zeki R 68":"30.7","Zeki R 40":"30.8","Zeki R 95":"44.6","Zeki L 74":"33.7","Zeki L 48":"84.8","Zeki L 28 Repl Avg.":"39.1","Zeki L 95":"11.2","Zeki L 40":"49.5","Zeki L 68":"40","Zeki T 74":"148","Zeki T 48":"138","Zeki T 28 Repl Avg.":"< 0","Zeki T 40":"136","Zeki T 68":"51.1","Zeki T 95":"37.7"},{"Metabolite":"TLCA","Zeki R 74":"1.93","Zeki R 28 Repl Avg.":"3.67","Zeki R 48":"1.13","Zeki R 68":"2.91","Zeki R 40":"1.69","Zeki R 95":"1.61","Zeki L 74":"1.4","Zeki L 48":"4.14","Zeki L 28 Repl Avg.":"3.21","Zeki L 95":"1.4","Zeki L 40":"3.67","Zeki L 68":"2.85","Zeki T 74":"8.05","Zeki T 48":"6.22","Zeki T 28 Repl Avg.":"0.66","Zeki T 40":"4.36","Zeki T 68":"3.26","Zeki T 95":"1.51"},{"Metabolite":"GDCA","Zeki R 74":"190","Zeki R 28 Repl Avg.":"100","Zeki R 48":"108","Zeki R 68":"273","Zeki R 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Avg.":"","Zeki L 95":"","Zeki L 40":"","Zeki L 68":"","Zeki T 74":"","Zeki T 48":"","Zeki T 28 Repl Avg.":"","Zeki T 40":"","Zeki T 68":"","Zeki T 95":""},{"Metabolite":"GDHCA screen","Zeki R 74":"","Zeki R 28 Repl Avg.":"","Zeki R 48":"","Zeki R 68":"","Zeki R 40":"","Zeki R 95":"","Zeki L 74":"","Zeki L 48":"","Zeki L 28 Repl Avg.":"","Zeki L 95":"","Zeki L 40":"","Zeki L 68":"","Zeki T 74":"","Zeki T 48":"","Zeki T 28 Repl Avg.":"","Zeki T 40":"","Zeki T 68":"","Zeki T 95":""},{"Metabolite":"MCA screen","Zeki R 74":"","Zeki R 28 Repl Avg.":"","Zeki R 48":"","Zeki R 68":"","Zeki R 40":"","Zeki R 95":"","Zeki L 74":"","Zeki L 48":"","Zeki L 28 Repl Avg.":"","Zeki L 95":"","Zeki L 40":"","Zeki L 68":"","Zeki T 74":"","Zeki T 48":"","Zeki T 28 Repl Avg.":"","Zeki T 40":"","Zeki T 68":"","Zeki T 95":""}],

"Metabolites":[{"metabolite_name":"CA"},{"metabolite_name":"CDCA"},{"metabolite_name":"TCA"},{"metabolite_name":"TCDCA"},{"metabolite_name":"GCA"},{"metabolite_name":"GCDCA"},{"metabolite_name":"DCA"},{"metabolite_name":"LCA"},{"metabolite_name":"TDCA"},{"metabolite_name":"TLCA"},{"metabolite_name":"GDCA"},{"metabolite_name":"GLCA"},{"metabolite_name":"GHDCA"},{"metabolite_name":"UDCA"},{"metabolite_name":"TUDCA"},{"metabolite_name":"GUDCA"},{"metabolite_name":"α-MCA"},{"metabolite_name":"β-MCA"},{"metabolite_name":"HCA screen"},{"metabolite_name":"T-ω-MCA"},{"metabolite_name":"T-α-MCA"},{"metabolite_name":"T-β-MCA"},{"metabolite_name":"THCA screen"},{"metabolite_name":"GHCA screen"},{"metabolite_name":"ω-MCA"},{"metabolite_name":"THDCA screen"},{"metabolite_name":"DHCA screen"},{"metabolite_name":"TDHCA"},{"metabolite_name":"GDHCA screen"},{"metabolite_name":"MCA screen"}]
}

}