Summary of study ST001091

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 PR000730. The data can be accessed directly via it's Project DOI: 10.21228/M89X1C This work is supported by NIH grant, U2C- DK119886.

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

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST001091
Study TitleAspirin Metabolomics in Colorectal Cancer Chemoprevention (part 1 - Colon)
Study TypeUntargeted high-resolution mass spectrometry profiling
Study SummarySubstantial evidence supports the effectiveness of aspirin for cancer chemoprevention in addition to its well established role in cardiovascular protection. In recent meta-analyses of randomized controlled trials in humans, daily aspirin use reduced incidence, metastasis and mortality from several common types of cancer, especially colorectal cancer. The mechanism(s) by which aspirin exerts an anticancer benefit is uncertain; numerous effects have been described involving both cyclooxygenase-dependent and -independent pathways. The goal of this research is to elucidate the key metabolic changes that are responsible for the anticancer effects of aspirin in humans using untargeted metabolomics analysis. Metabolomics, or global metabolite profiling, is an emerging discipline that has the potential to transform the study of pharmaceutical agents. Our innovative approach will use high-resolution mass spectroscopy to detect thousands of metabolites in blood plasma and normal colon mucosa biopsies that were collected from participants in the Aspirin/Folate Polyp Prevention Study, a randomized, double-blind, placebo-controlled trial of aspirin and/or folic acid supplementation for the prevention of colorectal adenomas. Participants in the trial were assigned with equal probability to three aspirin treatment arms (placebo, 81 mg, or 325 mg daily). Over the three-year treatment period, 81 mg/day of aspirin reduced the risk of adenomas, whereas the 325 mg/day dose had less effect. The aims of the current proposal are to identify metabolomic signatures, including specific metabolites and metabolic pathways, that are associated with aspirin treatment in blood and normal colon mucosal tissue of participants after three years of randomized aspirin treatment; and then to assess the associations of these metabolic signatures with adenoma risk and whether they mediate the reductions in risk due to 81 mg/day aspirin treatment. We will prioritize metabolites for study by evaluating metabolite levels in patients from the placebo and treatment arms while controlling the false discovery rate, use correlation analysis to enhance identification of relevant metabolic modules associated with these prioritized metabolites, and apply pathway mapping with post-hoc application of ion dissociation spectroscopy to representative metabolites to confirm pathway identification. Because aspirin is a multifunctional drug that is thought to modify numerous pathways with potential roles in carcinogenesis, a global discovery-based metabolomics approach is the best way to identify its key activities. The public health significance of this work is substantial because understanding the mechanism of aspirin’s anticancer effects is key to optimizing its use and to the development of novel drugs targeting the metabolic pathways identified.
Institute
Emory University
DepartmentSchool of Medicine
LaboratoryClincal Biomarkers Laboratory
Last NameUppal
First NameKaran
Address615 Michael Street, Atlanta, GA, 30322, USA
Emailkuppal2@emory.edu
Phone(404) 727 5027
Submit Date2018-09-05
Num Groups3
Total Subjects325
Num Males214
Num Females111
Study CommentsBoth pooled colon tissue samples and Clinical Biomarker Laboratory pooled plasma samples were used
Analysis Type DetailLC-MS
Release Date2019-09-23
Release Version1
Karan Uppal Karan Uppal
https://dx.doi.org/10.21228/M89X1C
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR000730
Project DOI:doi: 10.21228/M89X1C
Project Title:Aspirin Metabolomics in Colorectal Cancer Chemoprevention (part 1 - Colon)
Project Type:NIH/NCI R01CA188038
Project Summary:Substantial evidence supports the effectiveness of aspirin for cancer chemoprevention in addition to its well established role in cardiovascular protection. In recent meta-analyses of randomized controlled trials in humans, daily aspirin use reduced incidence, metastasis and mortality from several common types of cancer, especially colorectal cancer. The mechanism(s) by which aspirin exerts an anticancer benefit is uncertain; numerous effects have been described involving both cyclooxygenase-dependent and -independent pathways. The goal of this research is to elucidate the key metabolic changes that are responsible for the anticancer effects of aspirin in humans using untargeted metabolomics analysis. Metabolomics, or global metabolite profiling, is an emerging discipline that has the potential to transform the study of pharmaceutical agents. Our innovative approach will use high-resolution mass spectroscopy to detect thousands of metabolites in blood plasma and normal colon mucosa biopsies that were collected from participants in the Aspirin/Folate Polyp Prevention Study, a randomized, double-blind, placebo-controlled trial of aspirin and/or folic acid supplementation for the prevention of colorectal adenomas. Participants in the trial were assigned with equal probability to three aspirin treatment arms (placebo, 81 mg, or 325 mg daily). Over the three-year treatment period, 81 mg/day of aspirin reduced the risk of adenomas, whereas the 325 mg/day dose had less effect. The aims of the current proposal are to identify metabolomic signatures, including specific metabolites and metabolic pathways, that are associated with aspirin treatment in blood and normal colon mucosal tissue of participants after three years of randomized aspirin treatment; and then to assess the associations of these metabolic signatures with adenoma risk and whether they mediate the reductions in risk due to 81 mg/day aspirin treatment. We will prioritize metabolites for study by evaluating metabolite levels in patients from the placebo and treatment arms while controlling the false discovery rate, use correlation analysis to enhance identification of relevant metabolic modules associated with these prioritized metabolites, and apply pathway mapping with post-hoc application of ion dissociation spectroscopy to representative metabolites to confirm pathway identification. Because aspirin is a multifunctional drug that is thought to modify numerous pathways with potential roles in carcinogenesis, a global discovery-based metabolomics approach is the best way to identify its key activities. The public health significance of this work is substantial because understanding the mechanism of aspirin’s anticancer effects is key to optimizing its use and to the development of novel drugs targeting the metabolic pathways identified.
Institute:Emory University
Department:School of Medicine
Laboratory:Clinical Biomarkers Laboratory
Last Name:Uppal
First Name:Karan
Address:615 Michael Street, Atlanta, GA, 30322, USA
Email:kuppal2@emory.edu
Phone:(404) 727 5027
Funding Source:NIH/NCI R01CA188038
Project Comments:Requested embargo date: 8/1/2019
Contributors:Dartmouth: Elizabeth Barry Leila Mott John Baron Christopher Amos Michael Passarelli Emory: Dean Jones Veronika Fedirko Karan Uppal Shuzhao Li Douglas Walker Yutong Jin Chunyu Ma

Subject:

Subject ID:SU001135
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Age Or Age Range:25-78 yr
Gender:Male and female

Factors:

Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Aspirin Dosage Sample.Matrix
SA07363840023179_2ASA 325 Colon Mucosal Tissue
SA07363940023368_2ASA 325 Colon Mucosal Tissue
SA07364040023269_2ASA 325 Colon Mucosal Tissue
SA07364140023355_1ASA 325 Colon Mucosal Tissue
SA07364240023369_1ASA 325 Colon Mucosal Tissue
SA07364340023334_1ASA 325 Colon Mucosal Tissue
SA07364440023364_2ASA 325 Colon Mucosal Tissue
SA07364540023332_2ASA 325 Colon Mucosal Tissue
SA07364640023357_2ASA 325 Colon Mucosal Tissue
SA07364740023329_1ASA 325 Colon Mucosal Tissue
SA07364840023293_1ASA 325 Colon Mucosal Tissue
SA07364940023321_1ASA 325 Colon Mucosal Tissue
SA07365040023296_1ASA 325 Colon Mucosal Tissue
SA07365140023308_1ASA 325 Colon Mucosal Tissue
SA07365240023303_1ASA 325 Colon Mucosal Tissue
SA07365340023352_2ASA 325 Colon Mucosal Tissue
SA07365440023317_1ASA 325 Colon Mucosal Tissue
SA07365540023305_1ASA 325 Colon Mucosal Tissue
SA07365640023315_1ASA 325 Colon Mucosal Tissue
SA07365740023348_1ASA 325 Colon Mucosal Tissue
SA07365840023357_1ASA 325 Colon Mucosal Tissue
SA07365940023364_1ASA 325 Colon Mucosal Tissue
SA07366040023203_2ASA 325 Colon Mucosal Tissue
SA07366140023368_1ASA 325 Colon Mucosal Tissue
SA07366240023280_2ASA 325 Colon Mucosal Tissue
SA07366340023358_2ASA 325 Colon Mucosal Tissue
SA07366440023351_2ASA 325 Colon Mucosal Tissue
SA07366540023332_1ASA 325 Colon Mucosal Tissue
SA07366640023352_1ASA 325 Colon Mucosal Tissue
SA07366740023195_2ASA 325 Colon Mucosal Tissue
SA07366840023351_1ASA 325 Colon Mucosal Tissue
SA07366940023358_1ASA 325 Colon Mucosal Tissue
SA07367040023339_1ASA 325 Colon Mucosal Tissue
SA07367140023196_2ASA 325 Colon Mucosal Tissue
SA07367240023185_2ASA 325 Colon Mucosal Tissue
SA07367340023359_2ASA 325 Colon Mucosal Tissue
SA07367440023359_1ASA 325 Colon Mucosal Tissue
SA07367540023294_1ASA 325 Colon Mucosal Tissue
SA07367640023324_1ASA 325 Colon Mucosal Tissue
SA07367740023388_2ASA 325 Colon Mucosal Tissue
SA07367840023395_2ASA 325 Colon Mucosal Tissue
SA07367940023278_1ASA 325 Colon Mucosal Tissue
SA07368040023216_2ASA 325 Colon Mucosal Tissue
SA07368140023222_2ASA 325 Colon Mucosal Tissue
SA07368240023280_1ASA 325 Colon Mucosal Tissue
SA07368340023269_1ASA 325 Colon Mucosal Tissue
SA07368440023238_2ASA 325 Colon Mucosal Tissue
SA07368540023262_1ASA 325 Colon Mucosal Tissue
SA07368640023397_2ASA 325 Colon Mucosal Tissue
SA07368740023231_2ASA 325 Colon Mucosal Tissue
SA07368840023406_2ASA 325 Colon Mucosal Tissue
SA07368940023212_2ASA 325 Colon Mucosal Tissue
SA07369040023275_1ASA 325 Colon Mucosal Tissue
SA07369140023252_1ASA 325 Colon Mucosal Tissue
SA07369240023260_1ASA 325 Colon Mucosal Tissue
SA07369340023265_1ASA 325 Colon Mucosal Tissue
SA07369440023262_2ASA 325 Colon Mucosal Tissue
SA07369540023288_1ASA 325 Colon Mucosal Tissue
SA07369640023394_2ASA 325 Colon Mucosal Tissue
SA07369740023384_2ASA 325 Colon Mucosal Tissue
SA07369840023387_2ASA 325 Colon Mucosal Tissue
SA07369940023278_2ASA 325 Colon Mucosal Tissue
SA07370040023331_1ASA 325 Colon Mucosal Tissue
SA07370140023318_1ASA 325 Colon Mucosal Tissue
SA07370240023326_1ASA 325 Colon Mucosal Tissue
SA07370340023292_1ASA 325 Colon Mucosal Tissue
SA07370440023277_1ASA 325 Colon Mucosal Tissue
SA07370540023291_1ASA 325 Colon Mucosal Tissue
SA07370640023255_1ASA 325 Colon Mucosal Tissue
SA07370740023374_2ASA 325 Colon Mucosal Tissue
SA07370840023391_2ASA 325 Colon Mucosal Tissue
SA07370940023379_2ASA 325 Colon Mucosal Tissue
SA07371040023268_1ASA 325 Colon Mucosal Tissue
SA07371140023267_1ASA 325 Colon Mucosal Tissue
SA07371240023398_2ASA 325 Colon Mucosal Tissue
SA07371340023339_2ASA 325 Colon Mucosal Tissue
SA07371440023288_2ASA 325 Colon Mucosal Tissue
SA07371540023321_2ASA 325 Colon Mucosal Tissue
SA07371640023457_1ASA 325 Colon Mucosal Tissue
SA07371740023449_1ASA 325 Colon Mucosal Tissue
SA07371840023447_1ASA 325 Colon Mucosal Tissue
SA07371940023448_1ASA 325 Colon Mucosal Tissue
SA07372040023192_2ASA 325 Colon Mucosal Tissue
SA07372140023296_2ASA 325 Colon Mucosal Tissue
SA07372240023453_1ASA 325 Colon Mucosal Tissue
SA07372340023452_1ASA 325 Colon Mucosal Tissue
SA07372440023434_1ASA 325 Colon Mucosal Tissue
SA07372540023293_2ASA 325 Colon Mucosal Tissue
SA07372640023431_1ASA 325 Colon Mucosal Tissue
SA07372740023415_1ASA 325 Colon Mucosal Tissue
SA07372840023190_2ASA 325 Colon Mucosal Tissue
SA07372940023197_2ASA 325 Colon Mucosal Tissue
SA07373040023425_1ASA 325 Colon Mucosal Tissue
SA07373140023445_1ASA 325 Colon Mucosal Tissue
SA07373240023417_1ASA 325 Colon Mucosal Tissue
SA07373340023171_2ASA 325 Colon Mucosal Tissue
SA07373440023294_2ASA 325 Colon Mucosal Tissue
SA07373540023144_2ASA 325 Colon Mucosal Tissue
SA07373640023160_2ASA 325 Colon Mucosal Tissue
SA07373740023145_2ASA 325 Colon Mucosal Tissue
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Collection:

Collection ID:CO001129
Collection Summary:Fresh frozen normal colon mucosal tissue samples were collected from participants in the Aspirin/Folate Polyp Prevention Study [1, 2] at the year three (end of treatment) colonoscopy. After the endoscope was advanced to the cecum, biopsy specimens were obtained from the mid-ascending colon (5 cm above the ileocecal valve). Immediately after the biopsies were taken, they were removed from the forceps, placed into 1.8 ml freezer tubes and immersed in liquid nitrogen or a dry ice/ethanol slurry until storage in a -70C freezer. The specimens were shipped on dry ice from the clinical centers to the Dartmouth biorepository storage facility and subsequently to the metabolomics analysis lab at Emory University. In addition, a colon tissue reference was generated using normal colon tissue removed at surgery from an anonymous donor. References 1. Baron JA, Cole BF, Sandler RS, et al. A randomized trial of aspirin to prevent colorectal adenomas. The New England journal of medicine. 2003;348(10):891-9. 2. Cole BF, Baron JA, Sandler RS, et al. Folic acid for the prevention of colorectal adenomas: a randomized clinical trial. JAMA : the journal of the American Medical Association. 2007;297(21):2351-9.
Sample Type:Colon mucosal tissue
Storage Conditions:Described in summary

Treatment:

Treatment ID:TR001149
Treatment Summary:Samples were received frozen in aliquouts of <250uL. Prior to analysis, samples were thawed and prepared for HRM analysis using the standard protocols described in the Sample Preparation section.

Sample Preparation:

Sampleprep ID:SP001142
Sampleprep Summary:Samples were prepared for metabolomics analysis using established methods(Johnson et al. (2010). Analyst; Go et al. (2015). Tox Sci). Prior to analysis, plasma aliquots were removed from storage at -80 degrees C and thawed on ice. Each cryotube was then vortexed briefly to ensure homogeneity, and 50 microliters was transferred to a clean microfuge tube. Immediately after, the samples were treated with 200 microliters of ice-cold LC-MS grade acetonitrile (Sigma Aldrich) containing 5 microliters of internal standard solution with eight stable isotopic chemicals selected to cover a range of chemical properties. Following addition of acetonitrile, samples were homogenized for a few seconds using an Active Motif EpiShear probe sonicator and equilibrated for 30 min on ice, upon which precipitated proteins were removed by centrifuge (14,000 rpm at 4 degrees C for 10 min). The resulting supernatant (100 microliters) was removed, added to a low volume autosampler vial and maintained at 4 degrees C until analysis (<22 h).
Sampleprep Protocol ID:HRM_SP_082016_01
Sampleprep Protocol Filename:EmoryUniversity_HRM_sample_preparation_082016_01
Sampleprep Protocol Comments:Date effective: 30 July 2016
Extraction Method:2:1 acetonitrile: sample followed by vortexing and centrifugation

Combined analysis:

Analysis ID AN001776 AN001777
Analysis type MS MS
Chromatography type HILIC Reversed phase
Chromatography system Thermo Dionex Ultimate 3000 Thermo Dionex Ultimate 3000
Column Waters XBridge Amide (50 x 2.1mm, 2.5um) Thermo Higgins C18 (50 x 2.1mm, 3um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Fusion Tribrid Orbitrap Thermo Fusion Tribrid Orbitrap
Ion Mode POSITIVE NEGATIVE
Units Peak area Peak area

Chromatography:

Chromatography ID:CH001255
Chromatography Summary:The HILIC column is operated parallel to reverse phase column for simultaneous analytical separation and column flushing through the use of a dual head HPLC pump equipped with 10-port and 6-port switching valves. During operation of "HILIC separation method, the MS is operated in positive ion mode and 10" microliters of sample is injected onto the HILIC column while the reverse phase column is flushing with wash solution. Flow rate is maintained at 0.35 mL/min "until 1.5 min, increased to 0.4 mL/min at 4 min and held for 1 min. Solvent A is" "100% LC-MS grade water, solvent B is 100% LC-MS grade acetonitrile and solvent C" is 2% formic acid (v/v) in LC-MS grade water. Initial mobile phase conditions "are 22.5% A, 75% B, 2.5% C hold for 1.5 min, with linear gradient to 77.5% A," "20% B, 2.5% C at 4 min, hold for 1 min, resulting in a total analytical run time" "of 5 min. During the flushing phase (reverse phase analytical separation), the" "HILIC column is equilibrated with a wash solution of 77.5% A, 20% B, 2.5% C."
Methods ID:2% formic acid in LC-MS grade water
Methods Filename:20160920_posHILIC120kres5min_ESI_c18negwash.meth
Chromatography Comments:Triplicate injections for each chromatography mode
Instrument Name:Thermo Dionex Ultimate 3000
Column Name:Waters XBridge Amide (50 x 2.1mm, 2.5um)
Column Temperature:60C
Flow Gradient:A= water, B= acetontrile, C= 2% formic acid in water; 22.5% A, 75% B, 2.5% C hold for 1.5 min, linear gradient to 77.5% A, 20% B, 2.5% C at 4 min, hold for 1 min
Flow Rate:0.35 mL/min for 1.5 min; linear increase to 0.4 mL/min at 4 min, hold for 1 min
Sample Injection:10 uL
Solvent A:LC-MS grade water
Solvent B:LC-MS grade acetonitrile
Analytical Time:5 min
Sample Loop Size:15 uL
Sample Syringe Size:100 uL
Chromatography Type:HILIC
  
Chromatography ID:CH001256
Chromatography Summary:The C18 column is operated parallel to the HILIC column for simultaneous analytical separation and column flushing through the use of a dual head HPLC pump equipped with 10-port and 6- port switching valves. During operation of the "C18 method, the MS is operated in negative ion mode and 10 μL of sample is" injected onto the C18 column while the HILIC column is flushing with wash "solution. Flow rate is maintained at 0.4 mL/min until 1.5 min, increased to 0.5" "mL/min at 2 min and held for 3 min. Solvent A is 100% LC-MS grade water, solvent" B is 100% LC-MS grade acetonitrile and solvent C is 10mM ammonium acetate in "LC-MS grade water. Initial mobile phase conditions are 60% A, 35% B, 5% C hold" "for 0.5 min, with linear gradient to 0% A, 95% B, 5% C at 1.5 min, hold for 3.5" "min, resulting in a total analytical run time of 5 min. During the flushing" "phase (HILIC analytical separation), the C18 column is equilibrated with a wash" "solution of 0% A, 95% B, 5% C until 2.5 min, followed by an equilibration" "solution of 60% A, 35% B, 5% C for 2.5 min."
Methods ID:10mM ammonium acetate in LC-MS grade water
Methods Filename:20160920_negC18120kres5min_ESI_HILICposwash.meth
Instrument Name:Thermo Dionex Ultimate 3000
Column Name:Thermo Higgins C18 (50 x 2.1mm, 3um)
Column Temperature:60C
Flow Rate:0.4 mL/min for 1.5 min; linear increase to 0.5 mL/min at 2 min held for 3 min
Sample Injection:10 uL
Solvent A:LC-MS grade water
Solvent B:LC-MS grade acetonitrile
Analytical Time:5 min
Sample Loop Size:15 uL
Sample Syringe Size:100 uL
Chromatography Type:Reversed phase

MS:

MS ID:MS001641
Analysis ID:AN001776
Instrument Name:Thermo Fusion Tribrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
Ion Mode:POSITIVE
Capillary Temperature:250C
Collision Gas:N2
Dry Gas Flow:45
Dry Gas Temp:150C
Mass Accuracy:< 3ppm
Spray Voltage:3500
Activation Parameter:5.00E+05
Activation Time:118ms
Interface Voltage:S-Lens RF level= 55
Resolution Setting:120000
Scanning Range:85-1275
  
MS ID:MS001642
Analysis ID:AN001777
Instrument Name:Thermo Fusion Tribrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
Ion Mode:NEGATIVE
Capillary Temperature:250C
Collision Gas:N2
Dry Gas Flow:45
Dry Gas Temp:150C
Mass Accuracy:< 3ppm
Spray Voltage:-4000
Activation Parameter:5.00E+05
Activation Time:118ms
Interface Voltage:S-Lens RF level= 55
Resolution Setting:120000
Scanning Range:85-1275
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