Summary of Study ST002748
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 PR001711. The data can be accessed directly via it's Project DOI: 10.21228/M8JD9D 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 | ST002748 |
| Study Title | HER2 overexpression initiates breast tumorigenesis non-cell autonomously by inducing oxidative stress in the tissue microenvironment |
| Study Summary | HER2 is a driver oncogene overexpressed in the majority of premalignant breast tumors known as ductal carcinoma in situ (DCIS). Due to their stemness features, breast cancer stem cells (BCSC) are considered the main drivers of breast tumor initiation and progression. Here, we used clinical samples and mouse models of HER2+ breast tumorigenesis to demonstrate that neither BCSCs nor their cell-of-origin express HER2/Neu in early-stage breast tumors. Instead, our results demonstrate that Neu overexpression results in the transformation of BCSCs in a non-cell autonomous manner via triggering DNA damage and somatic mutagenesis in their Neu-negative cell-of-origin. This is caused by the increased oxidative stress in the tissue microenvironment generated by altered energy metabolism and increased reactive oxygen species level in Neu-overexpressing mammary ducts. Therefore, our findings illustrate a previously unrecognized mechanism of HER2-induced breast tumor initiation, which may have an impact on future preventive treatments for patients with HER2+ DCIS. |
| Institute | The University of Manchester |
| Department | Manchester Breast Centre |
| Laboratory | Ahmet Ucar Lab |
| Last Name | Ucar |
| First Name | Ahmet |
| Address | Oxford Road, Manchester, M13 9PL, UK. |
| ahmet.ucar@manchester.ac.uk | |
| Phone | +44 (0)161 3067116 |
| Submit Date | 2023-06-26 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | wiff |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-07-10 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001711 |
| Project DOI: | doi: 10.21228/M8JD9D |
| Project Title: | HER2 related MMTV-Neu mammary ducts studies |
| Project Type: | MS quantitative analysis |
| Project Summary: | Metabolomics analysis of intact mammary ducts. HER2 is a driver oncogene overexpressed in the majority of premalignant breast tumors known as ductal carcinoma in situ (DCIS). Due to their stemness features, breast cancer stem cells (BCSC) are considered the main drivers of breast tumor initiation and progression. Here, we used clinical samples and mouse models of HER2+ breast tumorigenesis to demonstrate that neither BCSCs nor their cell-of-origin express HER2/Neu in early-stage breast tumors. Instead, our results demonstrate that Neu overexpression results in the transformation of BCSCs in a non-cell autonomous manner via triggering DNA damage and somatic mutagenesis in their Neu-negative cell-of-origin. This is caused by the increased oxidative stress in the tissue microenvironment generated by altered energy metabolism and increased reactive oxygen species level in Neu-overexpressing mammary ducts. Therefore, our findings illustrate a previously unrecognized mechanism of HER2-induced breast tumor initiation, which may have an impact on future preventive treatments for patients with HER2+ DCIS. |
| Institute: | The University of Manchester |
| Department: | Faculty of Biology, Medicine and Health |
| Laboratory: | Ahmet Ucar Lab |
| Last Name: | Ou |
| First Name: | Yaqing |
| Address: | Oxford Road, Manchester, Great Manchester, M13 9PL, United Kingdom |
| Email: | yaqing.ou@manchester.ac.uk |
| Phone: | 07579759914 |
| Contributors: | Sevim B. Gurler, Oliver Wagstaff, Lili Dimitrova, Fuhui Chen, Robert Pedley, William Weston, Ian Donaldson, Brian A. Telfer, David Novo, Kyriaki Pavlou, George Taylor, Yaqing Ou, Kaye J. Williams, Andrew Gilmore, Keith Brennan, Ahmet Ucar |
Subject:
| Subject ID: | SU002855 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Age Or Age Range: | 8-weeks old |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype | Treatment |
|---|---|---|---|
| SA289304 | 407 668 | Mutant | MMTV-NIC |
| SA289305 | 407 667 | Mutant | MMTV-NIC |
| SA289306 | 407 663 | Mutant | MMTV-NIC |
| SA289307 | 407 662 | Mutant | MMTV-NIC |
| SA289308 | 407 661 | Wild-type | Control |
| SA289309 | 407 669 | Wild-type | Control |
| SA289310 | 407 660 | Wild-type | Control |
| Showing results 1 to 7 of 7 |
Collection:
| Collection ID: | CO002848 |
| Collection Summary: | No:4 mammary glands were dissected and spread on sterile microscope slides. Lymph nodes were removed, and the glands were cut into thin horizontal stripes before digesting them with Collagenase A (#11088793001, Roche) at 37°C for 3-4 hours. The ductal organoids were then separated from single cells using a 100-um mesh and collected into a tube by flushing them with medium. |
| Sample Type: | Epithelial cells |
Treatment:
| Treatment ID: | TR002864 |
| Treatment Summary: | The mouse model of HER2+ breast cancer, MMTV-Neu-IRES-Cre (MMTV-NIC), is used as the treatment group. |
Sample Preparation:
| Sampleprep ID: | SP002861 |
| Sampleprep Summary: | Ductal organoids were centrifuged, tissue pellets were snap-frozen in liquid nitrogen and then resuspended in 50% Methanol to incubate overnight at 4 °C. Next day, samples were centrifuged at 20,000 x g for 3 min and the top 50 µl supernatant was transferred to a glass autosampler vial with 300 µl insert and capped. Quality control samples were made by pooling 10 µl from each sample. |
Combined analysis:
| Analysis ID | AN004458 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo-Fisher Ultimate 3000 HPLC |
| Column | Agilent Poroshell 120 HILIC-Z |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6600 QTOF |
| Ion Mode | NEGATIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH003347 |
| Chromatography Summary: | Liquid chromatography-mass spectrometry analysis was performed using a Thermo-Fisher Ultimate 3000 HPLC system consisting of an HPG-3400RS high pressure gradient pump, TCC 3000SD column compartment and WPS 3000 Autosampler, coupled to a SCIEX 6600 TripleTOF Q-TOF mass spectrometer with TurboV ion source. The system was controlled by SCIEX Analyst 1.7.1, DCMS Link and Chromeleon Xpress software. |
| Instrument Name: | Thermo-Fisher Ultimate 3000 HPLC |
| Column Name: | Agilent Poroshell 120 HILIC-Z |
| Column Temperature: | 40 |
| Flow Gradient: | flow rate of 0.25 ml/min, starting at 96 % B for 2 minutes, ramping to 65 % B over 20 min, hold at 65 % B for 2 min, then back to 96 % B |
| Flow Rate: | 0.25 ml/min |
| Solvent A: | 100% water; 10 mM ammonium acetate adjusted to pH 9 with ammonium hydroxide and 20 µM medronic acid |
| Solvent B: | 85% acetonitrile/15% water; 10 mM ammonium acetate adjusted to pH 9 with ammonium hydroxide and 20 µM medronic acid |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004205 |
| Analysis ID: | AN004458 |
| Instrument Name: | Agilent 6600 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | NEGATIVE |
