Summary of Study ST002354
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 PR001511. The data can be accessed directly via it's Project DOI: 10.21228/M8CQ6T 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.
| Study ID | ST002354 |
| Study Title | Combination of TP-252 and Naproxen elicit tumor protective Eicosanoid changes. |
| Study Summary | Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the United States. Patients with the genetic disorder Familial Adenomatous Polyposis (FAP) develop hundreds to thousands of polyps that unless removed by prophylactic colectomy will progress to CRC at an early age. Non-steroidal anti-inflammatory drugs (NSAIDs) and -3 marine polyunsaturated fatty acids (PUFA), such as eicosapentaenoic acid (EPA), have been evaluated for their chemopreventive potential in delaying the onset of CRC in high-risk patients. In this study, we determined whether the NSAID, naproxen, alone or in combination with a chemically-stable form of EPA (TP-252), affects tumor formation in the ApcPirc rat model. When compared to control diet, animals fed naproxen or HD TP-252 had 66%, and 82% fewer tumors respectively. However, when fed a combination of naproxen and HD TP-252, animals exhibited a 95% reduction in tumor formation and a 98% reduction in tumor volume, respectively. To elucidate potential mechanisms of tumor protection, a comprehensive, targeted lipidomic analysis was performed on colonic mucosa to determine changes in eicosanoid metabolism. Animals receiving TP-252 alone or in combination with naproxen had significantly reduced mucosal levels of pro-inflammatory -6 eicosanoids (PGE2, 5-HETE, and 14,15-DiHETrE), along with a simultaneous increase in anti-inflammatory EPA-derived -3 eicosanoids. Our colonic mucosal lipidomic analysis also uncovered several potential pharmacodynamic (PD) lipid biomarkers, including resolvin E2, 9-HEPE, 12-HEPE and 18-HEPE, that were increased in both the tissue and plasma of rats receiving TP-252 and were significantly correlated with tumor protection. Further studies with this drug combination should be focused on dose optimization and the role of EPA-derived lipid mediators in CRC initiation and progression. |
| Institute | UConn Health |
| Last Name | Beach |
| First Name | Ryan |
| Address | 263 Farmington Avenue, Farmington CT 06030 |
| rbeach@uchc.edu | |
| Phone | 860-679-8703 |
| Submit Date | 2022-11-21 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | wiff |
| Analysis Type Detail | LC-MS |
| Release Date | 2022-12-22 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN003842 | AN003843 | AN003844 | AN003845 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | Reversed phase | Reversed phase |
| Chromatography system | Prominence XR | Prominence XR | Prominence XR | Prominence XR |
| Column | Targa C8, 2x10 mm, 5μ | Luna C18 (150 x 2.1mm,3um) | Targa C8, 2x10 mm, 5μ | Luna C18 (150 x 2.1mm,3um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | QTRAP | QTRAP | QTRAP | QTRAP |
| MS instrument name | ABI Sciex 5500 QTrap | ABI Sciex 5500 QTrap | ABI Sciex 5500 QTrap | ABI Sciex 5500 QTrap |
| Ion Mode | NEGATIVE | NEGATIVE | NEGATIVE | NEGATIVE |
| Units | ng/g tissue | mg/mL | ng/mL | mg/g tissue |
Chromatography:
| Chromatography ID: | CH002844 |
| Chromatography Summary: | HPLC was performed on a Prominence XR system (Shimadzu) using Luna C18 (3μ, 2.1x150 mm) column. The mobile phase consisted of a gradient between A: methanol-water-acetonitrile (10:85:5 v/v) and B: methanol-water-acetonitrile (90:5:5 v/v), both containing 0.1% ammonium acetate. The gradient program with respect to the composition of B was as follows: 0-1 min, 50%; 1-8 min, 50-80%; 8-15 min, 80-95%; and 15-17 min, 95%. The flow rate was 0.2 ml/min. The HPLC eluate was directly introduced to ESI source of QTRAP5500 mass analyzer (ABSCIEX) in the negative ion mode with following conditions: Curtain gas: 35 psi, GS1: 35 psi, GS2: 65 psi, Temperature: 600 ˚C, Ion Spray Voltage: -1500 V, Collision gas: low, Declustering Potential: -60 V, and Entrance Potential: -7 V. The eluate was monitored by Multiple Reaction Monitoring method to detect unique molecular ion – daughter ion combinations for each of the lipid mediators using a scheduled MRM around the expected retention time for each compound. Optimized Collisional Energies (18 – 35 eV) and Collision Cell Exit Potentials (7 – 10 V) were used for each MRM transition. Spectra of each peak detected in the scheduled MRM were recorded using Enhanced Product Ion scan to confirm the structural identity. The data was collected using Analyst 1.6.2 software and the MRM transition chromatograms were quantitated by MultiQuant software (both from ABSCIEX). The internal standard signals in each chromatogram were used for normalization, recovery, as well as relative quantitation of each analyte. |
| Instrument Name: | Prominence XR |
| Column Name: | Targa C8, 2x10 mm, 5μ |
| Flow Rate: | 0.25ml/min |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH002845 |
| Chromatography Summary: | HPLC was performed on a Prominence XR system (Shimadzu) using Luna C18 (3μ, 2.1x150 mm) column. The mobile phase consisted of a gradient between A: methanol-water-acetonitrile (10:85:5 v/v) and B: methanol-water-acetonitrile (90:5:5 v/v), both containing 0.1% ammonium acetate. The gradient program with respect to the composition of B was as follows: 0-1 min, 50%; 1-8 min, 50-80%; 8-15 min, 80-95%; and 15-17 min, 95%. The flow rate was 0.2 ml/min. The HPLC eluate was directly introduced to ESI source of QTRAP5500 mass analyzer (ABSCIEX) in the negative ion mode with following conditions: Curtain gas: 35 psi, GS1: 35 psi, GS2: 65 psi, Temperature: 600 ˚C, Ion Spray Voltage: -1500 V, Collision gas: low, Declustering Potential: -60 V, and Entrance Potential: -7 V. The eluate was monitored by Multiple Reaction Monitoring method to detect unique molecular ion – daughter ion combinations for each of the lipid mediators using a scheduled MRM around the expected retention time for each compound. Optimized Collisional Energies (18 – 35 eV) and Collision Cell Exit Potentials (7 – 10 V) were used for each MRM transition. Spectra of each peak detected in the scheduled MRM were recorded using Enhanced Product Ion scan to confirm the structural identity. The data was collected using Analyst 1.6.2 software and the MRM transition chromatograms were quantitated by MultiQuant software (both from ABSCIEX). The internal standard signals in each chromatogram were used for normalization, recovery, as well as relative quantitation of each analyte. |
| Instrument Name: | Prominence XR |
| Column Name: | Luna C18 (150 x 2.1mm,3um) |
| Flow Rate: | 0.2ml/min |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH002846 |
| Chromatography Summary: | HPLC was performed on a Prominence XR system (Shimadzu) using Luna C18 (3μ, 2.1x150 mm) column. The mobile phase consisted of a gradient between A: methanol-water-acetonitrile (10:85:5 v/v) and B: methanol-water-acetonitrile (90:5:5 v/v), both containing 0.1% ammonium acetate. The gradient program with respect to the composition of B was as follows: 0-1 min, 50%; 1-8 min, 50-80%; 8-15 min, 80-95%; and 15-17 min, 95%. The flow rate was 0.2 ml/min. The HPLC eluate was directly introduced to ESI source of QTRAP5500 mass analyzer (ABSCIEX) in the negative ion mode with following conditions: Curtain gas: 35 psi, GS1: 35 psi, GS2: 65 psi, Temperature: 600 ˚C, Ion Spray Voltage: -1500 V, Collision gas: low, Declustering Potential: -60 V, and Entrance Potential: -7 V. The eluate was monitored by Multiple Reaction Monitoring method to detect unique molecular ion – daughter ion combinations for each of the lipid mediators using a scheduled MRM around the expected retention time for each compound. Optimized Collisional Energies (18 – 35 eV) and Collision Cell Exit Potentials (7 – 10 V) were used for each MRM transition. Spectra of each peak detected in the scheduled MRM were recorded using Enhanced Product Ion scan to confirm the structural identity. The data was collected using Analyst 1.6.2 software and the MRM transition chromatograms were quantitated by MultiQuant software (both from ABSCIEX). The internal standard signals in each chromatogram were used for normalization, recovery, as well as relative quantitation of each analyte. |
| Instrument Name: | Prominence XR |
| Column Name: | Targa C8, 2x10 mm, 5μ |
| Flow Rate: | 0.25ml/min |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH002847 |
| Chromatography Summary: | HPLC was performed on a Prominence XR system (Shimadzu) using Luna C18 (3μ, 2.1x150 mm) column. The mobile phase consisted of a gradient between A: methanol-water-acetonitrile (10:85:5 v/v) and B: methanol-water-acetonitrile (90:5:5 v/v), both containing 0.1% ammonium acetate. The gradient program with respect to the composition of B was as follows: 0-1 min, 50%; 1-8 min, 50-80%; 8-15 min, 80-95%; and 15-17 min, 95%. The flow rate was 0.2 ml/min. The HPLC eluate was directly introduced to ESI source of QTRAP5500 mass analyzer (ABSCIEX) in the negative ion mode with following conditions: Curtain gas: 35 psi, GS1: 35 psi, GS2: 65 psi, Temperature: 600 ˚C, Ion Spray Voltage: -1500 V, Collision gas: low, Declustering Potential: -60 V, and Entrance Potential: -7 V. The eluate was monitored by Multiple Reaction Monitoring method to detect unique molecular ion – daughter ion combinations for each of the lipid mediators using a scheduled MRM around the expected retention time for each compound. Optimized Collisional Energies (18 – 35 eV) and Collision Cell Exit Potentials (7 – 10 V) were used for each MRM transition. Spectra of each peak detected in the scheduled MRM were recorded using Enhanced Product Ion scan to confirm the structural identity. The data was collected using Analyst 1.6.2 software and the MRM transition chromatograms were quantitated by MultiQuant software (both from ABSCIEX). The internal standard signals in each chromatogram were used for normalization, recovery, as well as relative quantitation of each analyte. |
| Instrument Name: | Prominence XR |
| Column Name: | Luna C18 (150 x 2.1mm,3um) |
| Flow Rate: | 0.2ml/min |
| Chromatography Type: | Reversed phase |
