Summary of Study ST004427
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 PR002798. The data can be accessed directly via it's Project DOI: 10.21228/M80C2F This work is supported by NIH grant, U2C- DK119886. See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST004427 |
| Study Title | Brief Pulses of High-Level Fluid Shear Stress Enhance Metastatic Potential and Rapidly Alter the Metabolism of Cancer Cells |
| Study Type | MS studies on PC-3 human prostate cancer and MDA-MB-231 human breast cancer cell lines |
| Study Summary | PC-3 and MDA-MB-231 cell lines were grown in complete media at 37C until 80% confluency. The cells were either held in suspension as static controls or exposed to fluid shear stress (FSS) as outlined in PMID:32187555. Once the final pulse of FSS was conducted the cells were then plunged into ice cold PBS, counted, centrifuged, and cell pellet flash frozen in liquid nitrogen. Cell pellets were stored at -80C until profiling was initiated. Three biological replicates and match controls were submitted for metabolomic profiling. The data from the metabolomic profiling revealed that immediately upon FSS exposure, cancer cells significantly utilize ATP to support the energetically costly events of cytoskeleton rearrangement. We additionally observed significant utilization in amino acids including serine, threonine, and methionine-- which suggests utilization of the one-carbon cycle. We identified a slight increase of lactate production in the PC-3 cells, which was not observed in the MDA-MB-231 cells. This suggests a potential difference in energy production methods between the two cancer types during FSS exposure. Herein, this metabolomic profile demonstrates immediate activation of metabolic pathways to support the energy demand that is required to resist destruction to FSS. |
| Institute | Eastern Virginia Medical School at Old Dominion University |
| Department | Department of Biomedical & Translational Sciences |
| Last Name | Henry |
| First Name | Michael |
| Address | 700 W Olney Rd, PO Box 1980, Norfolk, VA, 23501 |
| mhenry@odu.edu | |
| Phone | 757-446-5619 |
| Submit Date | 2025-07-08 |
| Num Groups | 4 |
| Total Subjects | 24 |
| Publications | Pope, AN; Moose, DL, et al. Brief Pulses of High-Level Fluid Shear Stress Enhance Metastatic Potential and Rapidly Alter the Metabolism of Cancer Cells. bioRxiv 2025 , doi: https://doi.org/10.1101/2025.04.09.647247 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-12-29 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002798 |
| Project DOI: | doi: 10.21228/M80C2F |
| Project Title: | Brief Pulses of High-Level Fluid Shear Stress Enhance Metastatic Potential and Rapidly Alter the Metabolism of Cancer Cells |
| Project Type: | GC-and LC-MS |
| Project Summary: | Circulating tumor cells (CTCs) face challenges to their survival including mechanical and oxidative stresses that are different from cancer cells in solid primary and metastatic tumors. The impact of adaptations to the fluid microenvironment of the circulation on the outcome of the metastatic cascade are not well understood. Here we find that cancer cells (PC-3, MDA-MB-231, Myc-CaP) exposed to brief pulses of high-level FSS exhibit enhanced invasiveness and anchorage-independent proliferation in vitro and enhanced metastatic colonization/tumor formation in vivo. Cancer cells exposed to FSS rapidly alter their metabolism in a manner that promotes survival by providing energy for cytoskeletal remodeling and contractility as well as reducing equivalents to counter oxidative stress associated with cell detachment. Thus, exposure to FSS may provide CTCs an unexpected survival benefit that promotes metastatic colonization. |
| Institute: | Eastern Virginia Medical School at Old Dominion University |
| Department: | Department of Biomedical & Translational Sciences |
| Laboratory: | Dr. Michael Henry |
| Last Name: | Henry |
| First Name: | Michael |
| Address: | 700 W Olney Rd, PO Box 1980, Norfolk, VA, 23501 |
| Email: | mhenry@odu.edu |
| Phone: | 757-446-5619 |
| Funding Source: | NIH |
| Publications: | doi: https://doi.org/10.1101/2025.04.09.647247 |
| Contributors: | Amanda N Pope, Devon L Moose, Eric B Taylor |
Subject:
| Subject ID: | SU004587 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
| Cell Biosource Or Supplier: | ATCC |
| Cell Passage Number: | P5 |
| Cell Counts: | 2E6 |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Factor |
|---|---|---|---|
| SA522767 | QC3_MDA_GC | blank | - |
| SA522768 | QC4_PC3_GC | blank | - |
| SA522769 | QC3_PC3_GC | blank | - |
| SA522770 | QC2_PC3_GC | blank | - |
| SA522771 | QC1_PC3_GC | blank | - |
| SA522772 | Process blank_PC3_GC | blank | - |
| SA522773 | PC3_Pyridine_blank_GC | blank | - |
| SA522774 | QC5_MDA_LC | blank | - |
| SA522775 | QC4_MDA_LC | blank | - |
| SA522776 | QC3_MDA_LC | blank | - |
| SA522777 | QC2_MDA_LC | blank | - |
| SA522778 | QC1_MDA_LC | blank | - |
| SA522779 | QC1_MDA_GC | blank | - |
| SA522780 | QC2_MDA_GC | blank | - |
| SA522781 | Blank5_PC3_LC | blank | - |
| SA522782 | Processing_Blank_MDA_GC | blank | - |
| SA522783 | Blank4_PC3_LC | blank | - |
| SA522784 | QC3_PC3_LC | blank | - |
| SA522785 | QC2_PC3_LC | blank | - |
| SA522786 | Blan3_PC3_LC | blank | - |
| SA522787 | QC1_PC3_LC | blank | - |
| SA522788 | Blank3_PC3_LC | blank | - |
| SA522789 | PB_PC3_LC | blank | - |
| SA522790 | Blank2_PC3_LC | blank | - |
| SA522791 | QC6_MDA_LC | blank | - |
| SA522792 | QC4_MDA_GC | blank | - |
| SA522793 | QC5_MDA_GC | blank | - |
| SA522794 | QC6_MDA_GC | blank | - |
| SA522795 | blank_MDA_GC | blank | - |
| SA522796 | Human_Plasma_Reference_MDA_GC | blank | - |
| SA522797 | Blank1_PC3_LC | blank | - |
| SA522735 | MDA_S4_GC | MDA-MB-231 cancer cells | FSS |
| SA522736 | MDA_S1_LC | MDA-MB-231 cancer cells | FSS |
| SA522737 | MDA_S2 | MDA-MB-231 cancer cells | FSS |
| SA522738 | MDA_S3_LC | MDA-MB-231 cancer cells | FSS |
| SA522739 | MDA_S4_LC | MDA-MB-231 cancer cells | FSS |
| SA522740 | MDA_S3_GC | MDA-MB-231 cancer cells | FSS |
| SA522741 | MDA_S2_GC | MDA-MB-231 cancer cells | FSS |
| SA522742 | MDA_S1_GC | MDA-MB-231 cancer cells | FSS |
| SA522743 | MDA_U2_GC | MDA-MB-231 cancer cells | unsheared |
| SA522744 | MDA_U2_LC | MDA-MB-231 cancer cells | unsheared |
| SA522745 | MDA_U4_GC | MDA-MB-231 cancer cells | unsheared |
| SA522746 | MDA_U3_GC | MDA-MB-231 cancer cells | unsheared |
| SA522747 | MDA_U1_LC | MDA-MB-231 cancer cells | unsheared |
| SA522748 | MDA_U1_GC | MDA-MB-231 cancer cells | unsheared |
| SA522749 | MDA_U4_LC | MDA-MB-231 cancer cells | unsheared |
| SA522750 | MDA_U3_LC | MDA-MB-231 cancer cells | unsheared |
| SA522751 | PC3_S3_LC | PC-3 cancer cells | FSS |
| SA522752 | PC3_S4 | PC-3 cancer cells | FSS |
| SA522753 | PC3_S3 | PC-3 cancer cells | FSS |
| SA522754 | PC3_S2 | PC-3 cancer cells | FSS |
| SA522755 | PC3_S1 | PC-3 cancer cells | FSS |
| SA522756 | PC3_S1_LC | PC-3 cancer cells | FSS |
| SA522757 | PC3_S2_LC | PC-3 cancer cells | FSS |
| SA522758 | PC3_S4_LC | PC-3 cancer cells | FSS |
| SA522759 | PC3_U4_LC | PC-3 cancer cells | unsheared |
| SA522760 | PC3_U3 | PC-3 cancer cells | unsheared |
| SA522761 | PC3_U1 | PC-3 cancer cells | unsheared |
| SA522762 | PC3_U2 | PC-3 cancer cells | unsheared |
| SA522763 | PC3_U4 | PC-3 cancer cells | unsheared |
| SA522764 | PC3_U3_LC | PC-3 cancer cells | unsheared |
| SA522765 | PC3_U2_LC | PC-3 cancer cells | unsheared |
| SA522766 | PC3_U1_LC | PC-3 cancer cells | unsheared |
| Showing results 1 to 63 of 63 |
Collection:
| Collection ID: | CO004580 |
| Collection Summary: | PC-3 and MDA-MB-231 cancer cells were grown to 80% in 37C at 5% CO2 and harvested using 0.05% trypsin. Cells were washed with PBS, resuspended in serum-free DMEM. Metabolomic profiling was performed on cells that were processed and snap frozen (using liquid nitrogen) immediately after they were exposed to 10 pulses of FSS or held in suspension as described in PMID: 32187555. Specifically, on the final FSS pulse, the cells were transferred into 10 mL of ice-cold PBS, the number of viable cells was determined, and the cells were centrifuged at 500g for 3 minutes at 4°C. Cell debris was removed from pellets by resuspension in ice-cold PBS, 2x106 cells were transferred into a new tube, the cells were pelleted again, and these washed cell pellets were snap frozen in liquid nitrogen. |
| Sample Type: | Cultured cells |
| Collection Method: | cells were collected in ice cold PBS, centrifuged, then snap frozen using liquid nitrogen |
| Storage Conditions: | -80℃ |
| Tissue Cell Quantity Taken: | 2x10^6 cells |
Treatment:
| Treatment ID: | TR004596 |
| Treatment Summary: | No additional drug or pharmacological treatments were performed on the PC-3 or MDA-MB-231 cells prior to FSS exposure. Both cell lines were either held in suspension or exposure to a syringe and needle pump apparatus for 10 pulses. |
Sample Preparation:
| Sampleprep ID: | SP004593 |
| Sampleprep Summary: | Metabolomic profiling was performed on cells that were processed and snap frozen (using liquid nitrogen) immediately after they were exposed to 10 pulses of FSS or held in suspension as described above. Specifically, on the final FSS pulse, the cells were transferred into 10 mL of ice-cold PBS, the number of viable cells was determined, and the cells were centrifuged at 500g for 3 minutes at 4°C. Cell debris was removed from pellets by resuspension in ice-cold PBS, 2x106 cells were transferred into a new tube, the cells were pelleted again, and these washed cell pellets were snap frozen. |
Combined analysis:
| Analysis ID | AN007405 | AN007406 |
|---|---|---|
| Chromatography ID | CH005610 | CH005611 |
| MS ID | MS007097 | MS007098 |
| Analysis type | MS | MS |
| Chromatography type | GC | HILIC |
| Chromatography system | Thermo Trace 1310 | Thermo Vanquish |
| Column | TraceGold TG-5SilMS column (0.25 um film thickness; 0.25mm ID; 30 m length). | SeQuant ZIC- pHILIC (150 x 2.1 mm, 5 μm) |
| MS Type | EI | ESI |
| MS instrument type | Single quadrupole | Orbitrap |
| MS instrument name | Thermo ISQ LT | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | UNSPECIFIED |
| Units | pmole/L | pmole/L |
Chromatography:
| Chromatography ID: | CH005610 |
| Methods Filename: | GC-MS_Method_(ISQ)_Cell_Pellets_01128.pdf |
| Instrument Name: | Thermo Trace 1310 |
| Column Name: | TraceGold TG-5SilMS column (0.25 um film thickness; 0.25mm ID; 30 m length). |
| Column Temperature: | 25°C |
| Flow Gradient: | n/a |
| Flow Rate: | 1.2 mL/min |
| Solvent A: | n/a |
| Solvent B: | n/a |
| Chromatography Type: | GC |
| Chromatography ID: | CH005611 |
| Methods Filename: | MetCore_LC-MS_Method_HILIC_082522_Cells.pdf |
| Instrument Name: | Thermo Vanquish |
| Column Name: | SeQuant ZIC- pHILIC (150 x 2.1 mm, 5 μm) |
| Column Temperature: | 25°C |
| Flow Gradient: | The gradient starts at 80% B and decreasing to 20% B over 20 minutes; returning to 80% B in 0.5 minutes; and held there for 7 minutes. (PMID: 28388410). |
| Flow Rate: | 0.150 mL/min |
| Solvent A: | 100% Water; 20 mM ammonium carbonate; 0.1% Ammonium Hydroxide (v/v), pH is ~9.1 |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS007097 |
| Analysis ID: | AN007405 |
| Instrument Name: | Thermo ISQ LT |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| MS Comments: | GC chromatographic separation was conducted on a Thermo Trace 1300 GC with a TraceGold TG-5SilMS column (0.25 um film thickness; 0.25 mm ID; 30 m length). The injection volume of 1 μL was used for all samples, blanks, and QCs. The GC was operated in split mode with the following settings: 20:1 split ratio; split flow: 24 μL/min, purge flow: 5 mL/min, Carrier mode: Constant Flow, Carrier flow rate: 1.2 mL/min). The GC inlet temperature was 250°C. The GC oven temperature gradient was as follows: 80°C for 3 minutes, ramped at 20°C/minute to a maximum temperature of 280°C, which was held for 8 minutes. The injection syringe was washed 3 times with pyridine between each sample. Metabolites were detected using a Thermo ISQ single quadrupole mass spectrometer. The data was acquired from 3.90 to 21.00 minutes in EI mode (70eV) by single ion monitoring (SIM). |
| Ion Mode: | POSITIVE |
| MS ID: | MS007098 |
| Analysis ID: | AN007406 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The mass spectrometer was operated in full-scan, polarity-switching mode from 1 to 20 minutes, with the spray voltage set to 3.0 kV, the heated capillary held at 275°C, and the HESI probe held at 350°C. The sheath gas flow was set to 40 units, the auxiliary gas flow was set to 15 units, and the sweep gas flow was set to 1 unit. MS data acquisition was performed in a range of m/z 70–1,000, with the resolution set at 70,000, the AGC target at 1 × 106, and the maximum injection time at 200 ms.(PMID: 28388410) Data Analysis Acquired LC-MS data were processed by Thermo Scientific TraceFinder 4.1 software, and metabolites were identified based on the University of Iowa Metabolomics Core facility standard-confirmed, inhouse library. NOREVA was used for signal drift correction (PMID: 28525573). Data were normalized to the sum of all the measured metabolite ions in that sample. Polarity switching was used as the ion mode. |
| Ion Mode: | UNSPECIFIED |
