Summary of Study ST000843

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000600. The data can be accessed directly via it's Project DOI: 10.21228/M83X2W This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

Study ID	ST000843
Study Title	Statin Immuno-Metabolomics in Asthma (part I)
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	University of California, Davis
Department	USDA Western Human Nutrition Research Center
Laboratory	Newman's Lab
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
Submit Date	2017-08-09
Raw Data Available	Yes
Raw Data File Type(s)	wiff
Analysis Type Detail	LC-MS
Release Date	2017-10-11
Release Version	1

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Project:

Project ID:	PR000600
Project DOI:	doi: 10.21228/M83X2W
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

Subject:

Subject ID:	SU000870
Subject Type:	Animal
Subject Species:	Macaca mulatta
Taxonomy ID:	9544
Species Group:	Mammal

Factors:

Subject type: Animal; Subject species: Macaca mulatta (Factor headings shown in green)

mb_sample_id	local_sample_id	Treatment	Lung	Time point (day)
SA046783	BAL-AZ-01	Control	Left Lung	-
SA046784	AZ 16	Control	Left Lung	-
SA046785	AZ 33	Control	Left Lung	-
SA046786	BAL-AZ-33	Control	Left Lung	-
SA046787	BAL-AZ-34	Control	Left Lung	12
SA046788	AZ 13	Control	Left Lung	12
SA046789	AZ 10	Control	Left Lung	12
SA046790	AZ 45	Control	Left Lung	12
SA046791	BAL-AZ-19	Control	Left Lung	12
SA046792	BAL-AZ-02	Control	Left Lung	12
SA046793	BAL-AZ-04	Control	Left Lung	8
SA046794	AZ 07	Control	Left Lung	8
SA046795	BAL-AZ-09	Control	Left Lung	8
SA046796	BAL-AZ-10	Control	Left Lung	8
SA046797	AZ 30	Control	Left Lung	8
SA046798	AZ 36	Control	Left Lung	8
SA046799	AZ 03	Control	Right Lung	-
SA046800	AZ 06	Control	Right Lung	-
SA046801	BAL-AZ-07	Control	Right Lung	-
SA046802	AZ 32	Control	Right Lung	-
SA046803	BAL-AZ-18	Control	Right Lung	-
SA046804	AZ 38	Control	Right Lung	12
SA046805	BAL-AZ-26	Control	Right Lung	12
SA046806	AZ 22	Control	Right Lung	12
SA046807	AZ 29	Control	Right Lung	12
SA046808	BAL-AZ-12	Control	Right Lung	12
SA046809	BAL-AZ-22	Control	Right Lung	12
SA046810	AZ 01	Control	Right Lung	8
SA046811	BAL-AZ-14	Control	Right Lung	8
SA046812	BAL-AZ-17	Control	Right Lung	8
SA046813	AZ 24	Control	Right Lung	8
SA046814	BAL-AZ-13	Control	Right Lung	8
SA046815	AZ 39	Control	Right Lung	8
SA046816	AZ 21	Pravastatin	Left Lung	-
SA046817	AZ 26	Pravastatin	Left Lung	-
SA046818	AZ 23	Pravastatin	Left Lung	12
SA046819	AZ 15	Pravastatin	Left Lung	12
SA046820	AZ 43	Pravastatin	Left Lung	12
SA046821	AZ 44	Pravastatin	Left Lung	8
SA046822	AZ 02	Pravastatin	Left Lung	8
SA046823	AZ 37	Pravastatin	Left Lung	8
SA046824	AZ 40	Pravastatin	Right Lung	-
SA046825	AZ 17	Pravastatin	Right Lung	-
SA046826	AZ 08	Pravastatin	Right Lung	-
SA046827	AZ 31	Pravastatin	Right Lung	12
SA046828	AZ 09	Pravastatin	Right Lung	12
SA046829	AZ 25	Pravastatin	Right Lung	12
SA046830	AZ 14	Pravastatin	Right Lung	8
SA046831	AZ 18	Pravastatin	Right Lung	8
SA046832	BAL-AZ-24	Simvastatin	Left Lung	-
SA046833	BAL-AZ-06	Simvastatin	Left Lung	-
SA046834	BAL-AZ-11	Simvastatin	Left Lung	-
SA046835	BAL-AZ-05	Simvastatin	Left Lung	12
SA046836	BAL-AZ-35	Simvastatin	Left Lung	12
SA046837	BAL-AZ-25	Simvastatin	Left Lung	12
SA046838	BAL-AZ-23	Simvastatin	Left Lung	8
SA046839	BAL-AZ-28	Simvastatin	Left Lung	8
SA046840	BAL-AZ-08	Simvastatin	Left Lung	8
SA046841	BAL-AZ-30	Simvastatin	Right Lung	-
SA046842	BAL-AZ-27	Simvastatin	Right Lung	-
SA046843	BAL-AZ-03	Simvastatin	Right Lung	-
SA046844	BAL-AZ-21	Simvastatin	Right Lung	12
SA046845	BAL-AZ-31	Simvastatin	Right Lung	12
SA046846	BAL-AZ-20	Simvastatin	Right Lung	12
SA046847	BAL-AZ-32	Simvastatin	Right Lung	8
SA046848	BAL-AZ-29	Simvastatin	Right Lung	8
SA046849	BAL-AZ-15	Simvastatin	Right Lung	8

Showing results 1 to 67 of 67

Showing results 1 to 67 of 67

Collection:

Collection ID:	CO000864
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. Lung wash fluid was collected at day 0, 8 and 12 of each treatment.
Sample Type:	Bronchoalveolar lavage

Treatment:

Treatment ID:	TR000884
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. Lung wash fluid was collected at day 0, 8 and 12 of each treatment.

Sample Preparation:

Sampleprep ID:	SP000877
Sampleprep Summary:	Oxylipins, endocannabinoids, and fatty acids were isolated by solid phase extraction on 60 mg Waters Oasis-HLB cartridges (Milford, MA), as previously described by Luria et al (1). Prior to extraction, cartridges were washed with 1 column volume ethyl acetate followed by 2 column volumes methanol and conditioned with 2 mL of 95:5 v/v water/methanol (MeOH) with 0.1% acetic acid. The column reservoir was spiked with 5 µL anti-oxidant solution, (0.2 mg/ml solution BHT/EDTA in 1:1 MeOH:water), and 10 μL 1000nM analytical surrogates (See Table 2 below for specific compounds). Sample aliquots (500 µL media) were then introduced to the column reservoir and diluted with 1 column volume wash solution (5% MeOH, 0.1% acetic acid) and allowed to gravity extract into tubes containing 10% bleach solution to decontaminate sample waste. SPE cartridges were dried by vacuum @ -7.5in Hg for 20 min. Analytes were then eluted by gravity with 0.5 mL MeOH, followed by 1.25 mL Ethyl Acetate, into 2 mL autosampler vials containing 10 µL 20% glycerol solution in MeOH. Eluent was dried by vacuum evaporation for 35 min, and residues were re-constituted with 100uL of 100 nM internal standard solution containing 1-cyclohexyl ureido,3-dodecanoic acid (CUDA) and 1-Phenyl 3-Hexadecanoic Acid Urea (PHAU), in 50:50 MeOH:ACN and let sit for 10 min at ambient temperature. Vials were then chilled 15 min on wet ice, and extracts were transferred to a centrifugal filter (0.1 µm Durapore, Millipore, Billerica, MA), centrifuged for 3 min at 6ºC at <4500g (rcf) and transferred to 150 uL glass inserts within 2 mL amber vials, and capped. Extracts were stored at -20ºC until analysis by UPLC-MS/MS. The internal standard was used to quantify the recovery of the deuterated extraction surrogates by ratio response.

Sampleprep ID:

SP000877

Sampleprep Summary:

Oxylipins, endocannabinoids, and fatty acids were isolated by solid phase extraction on 60 mg Waters Oasis-HLB cartridges (Milford, MA), as previously described by Luria et al (1). Prior to extraction, cartridges were washed with 1 column volume ethyl acetate followed by 2 column volumes methanol and conditioned with 2 mL of 95:5 v/v water/methanol (MeOH) with 0.1% acetic acid. The column reservoir was spiked with 5 µL anti-oxidant solution, (0.2 mg/ml solution BHT/EDTA in 1:1 MeOH:water), and 10 μL 1000nM analytical surrogates (See Table 2 below for specific compounds). Sample aliquots (500 µL media) were then introduced to the column reservoir and diluted with 1 column volume wash solution (5% MeOH, 0.1% acetic acid) and allowed to gravity extract into tubes containing 10% bleach solution to decontaminate sample waste. SPE cartridges were dried by vacuum @ -7.5in Hg for 20 min. Analytes were then eluted by gravity with 0.5 mL MeOH, followed by 1.25 mL Ethyl Acetate, into 2 mL autosampler vials containing 10 µL 20% glycerol solution in MeOH. Eluent was dried by vacuum evaporation for 35 min, and residues were re-constituted with 100uL of 100 nM internal standard solution containing 1-cyclohexyl ureido,3-dodecanoic acid (CUDA) and 1-Phenyl 3-Hexadecanoic Acid Urea (PHAU), in 50:50 MeOH:ACN and let sit for 10 min at ambient temperature. Vials were then chilled 15 min on wet ice, and extracts were transferred to a centrifugal filter (0.1 µm Durapore, Millipore, Billerica, MA), centrifuged for 3 min at 6ºC at <4500g (rcf) and transferred to 150 uL glass inserts within 2 mL amber vials, and capped. Extracts were stored at -20ºC until analysis by UPLC-MS/MS. The internal standard was used to quantify the recovery of the deuterated extraction surrogates by ratio response.

Combined analysis:

Analysis ID	AN001364	AN001365
Analysis type	MS	MS
Chromatography type	Reversed phase	Reversed phase
Chromatography system	Waters Acquity	Waters Acquity
Column	Waters Acquity BEH C18 (100 x 2mm,1.7um)	Waters Acquity BEH C18 (100 x 2mm,1.7um)
MS Type	ESI	ESI
MS instrument type	Triple quadrupole	Triple quadrupole
MS instrument name	ABI Sciex 6500 QTrap	ABI Sciex 6500 QTrap
Ion Mode	NEGATIVE	POSITIVE
Units	Concentration (nM)	Concentration (nM)

Chromatography:

Chromatography ID:	CH000952
Instrument Name:	Waters Acquity
Column Name:	Waters Acquity BEH C18 (100 x 2mm,1.7um)
Column Temperature:	60 °C
Flow Gradient:	See protocol/methods file
Flow Rate:	0.25 mL/min
Internal Standard:	See protocol/methods file
Retention Time:	See protocol/methods file
Sample Injection:	5 µL
Solvent A:	100% water; 0.1% acetic acid
Solvent B:	90% acetonitrile/ 10% isopropanol
Analytical Time:	20 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
Chromatography Type:	Reversed phase

MS:

MS ID:	MS001256
Analysis ID:	AN001364
Instrument Name:	ABI Sciex 6500 QTrap
Instrument Type:	Triple quadrupole
MS Type:	ESI
Ion Mode:	NEGATIVE

MS ID:	MS001257
Analysis ID:	AN001365
Instrument Name:	ABI Sciex 6500 QTrap
Instrument Type:	Triple quadrupole
MS Type:	ESI
Ion Mode:	POSITIVE