Summary of Study ST001984

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 PR001260. The data can be accessed directly via it's Project DOI: 10.21228/M8V401 This work is supported by NIH grant, U2C- DK119886.

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

Perform statistical analysis | Show all samples | Show named metabolites | Download named metabolite data
Download mwTab file (text) | Download mwTab file(JSON)

Study ID	ST001984
Study Title	Metabolic adaptations in an endocrine-related breast cancer mouse model unveil potential markers of tumor response to hormonal therapy
Study Type	case - control study
Study Summary	Breast cancer (BC) is the most common type of cancer in women and, in most cases, it is hormone-dependent (HD), thus relying on ovarian hormone activation of intracellular receptors to stimulate tumor growth. Endocrine therapy (ET) aimed at preventing hormone receptor activation is the primary treatment strategy, however, about half of the patients, develop resistance in time. This involves the development of hormone independent tumors that initially are ET-responsive (HI), which may subsequently become resistant (HIR). The mechanisms that promote the conversion of HI to HIR tumors are varied and not completely understood. The aim of this work was to characterize the metabolic adaptations accompanying this conversion through the analysis of the polar metabolomes of tumor tissue and non-compromised mammary gland from mice implanted subcutaneously with HD, HI and HIR tumors from a medroxyprogesterone acetate (MPA)-induced BC mouse model. This was carried out by nuclear magnetic resonance (NMR) spectroscopy of tissue polar extracts and data mining through multivariate and univariate statistical analysis. Initial results unveiled marked changes between global tumor profiles and non-compromised mammary gland tissues, as expected. More importantly, specific metabolic signatures were found to accompany progression from HD, through HI and to HIR tumors, impacting on amino acids, nucleotides, membrane percursors and metabolites related to oxidative stress protection mechanisms. For each transition, sets of polar metabolites are advanced as potential markers of progression, including acquisition of resistance to ET. Putative biochemical interpretation of such signatures are proposed and discussed.
Institute	University of Aveiro
Department	Chemistry
Laboratory	Metabolomics
Last Name	Silva
First Name	Ana
Address	Campus Universitário de Santiago, 3810-193
Email	anarita.asilva@ua.pt
Phone	234370200
Submit Date	2021-10-06
Num Groups	8
Total Subjects	48
Num Females	48
Study Comments	For this study 48 female 2-month old Balb/c mice were used
Analysis Type Detail	NMR
Release Date	2021-12-01
Release Version	1

Select appropriate tab below to view additional metadata details:

Sample Preparation:

Sampleprep ID:	SP002071
Sampleprep Summary:	Both tumor and MG tissue were ground using a pestle and mortar, while kept in liquid nitrogen. All tissue samples (average weight of 50 mg) were extracted using methanol: chloroform: water (1:1:0.75) and the polar phase was separated for analysis. In brief, ground tissue samples were transferred to an eppendorf tube, followed by the addition of 500 µL of cold 80% methanol, 400µL of cold chloroform and 200 µL of cold Mili-Q water, and vortex homogenisation for 60 s. Samples were left to rest on ice for 10 minutes and then centrifuged (8,000 rpm, 5 min, 23 ºC). Polar phases were separated, vacuum-dried and stored at -80ºC until analysis. At the time of NMR acquisition, the dried aqueous extracts were suspended in 600 µL of 100 mM sodium phosphate buffer (pH 7.4, in D2O containing 0.25% 3-(trimethylsilyl)-propionic-2,2,3,3-d4 acid (TSP) for chemical shift referencing), homogenized, and 550 µL were transferred to 5mm NMR tubes. All NMR spectra were acquired on a Bruker AVANCE III spectrometer (Rheinstetten, Germany) operating at 500.13 MHz for proton. Standard 1D 1H NMR spectra of aqueous extracts were recorded at 298 K with water presaturation, using the “noesypr1d” pulse program (Bruker library), with 2.34 s acquisition time, 2 s relaxation delay, 512 scans, 7002.801 Hz spectral width, and 32 k data points. Each free-induction decay was zero-filled to 64 k points and multiplied by a 0.3 Hz exponential function prior to Fourier transformation. After acquisition, spectra were manually phased, baseline-corrected, and chemical-shift referenced to TSP. For selected samples, two-dimensional NMR spectra, namely 1H-1H TOCSY (Total Correlation Spectroscopy) and 1H-13C HSQC (Heteronuclear Single Quantum Coherence) spectra were also recorded to support spectral assignment. Peak assignment was also based on comparison with data available on the Bruker BBIOREFCODE spectral database and the human metabolome database (HMDB), as well as on existing literature.

Sampleprep ID:

SP002071

Sampleprep Summary:

Both tumor and MG tissue were ground using a pestle and mortar, while kept in liquid nitrogen. All tissue samples (average weight of 50 mg) were extracted using methanol: chloroform: water (1:1:0.75) and the polar phase was separated for analysis. In brief, ground tissue samples were transferred to an eppendorf tube, followed by the addition of 500 µL of cold 80% methanol, 400µL of cold chloroform and 200 µL of cold Mili-Q water, and vortex homogenisation for 60 s. Samples were left to rest on ice for 10 minutes and then centrifuged (8,000 rpm, 5 min, 23 ºC). Polar phases were separated, vacuum-dried and stored at -80ºC until analysis. At the time of NMR acquisition, the dried aqueous extracts were suspended in 600 µL of 100 mM sodium phosphate buffer (pH 7.4, in D2O containing 0.25% 3-(trimethylsilyl)-propionic-2,2,3,3-d4 acid (TSP) for chemical shift referencing), homogenized, and 550 µL were transferred to 5mm NMR tubes. All NMR spectra were acquired on a Bruker AVANCE III spectrometer (Rheinstetten, Germany) operating at 500.13 MHz for proton. Standard 1D 1H NMR spectra of aqueous extracts were recorded at 298 K with water presaturation, using the “noesypr1d” pulse program (Bruker library), with 2.34 s acquisition time, 2 s relaxation delay, 512 scans, 7002.801 Hz spectral width, and 32 k data points. Each free-induction decay was zero-filled to 64 k points and multiplied by a 0.3 Hz exponential function prior to Fourier transformation. After acquisition, spectra were manually phased, baseline-corrected, and chemical-shift referenced to TSP. For selected samples, two-dimensional NMR spectra, namely 1H-1H TOCSY (Total Correlation Spectroscopy) and 1H-13C HSQC (Heteronuclear Single Quantum Coherence) spectra were also recorded to support spectral assignment. Peak assignment was also based on comparison with data available on the Bruker BBIOREFCODE spectral database and the human metabolome database (HMDB), as well as on existing literature.