Summary of Study ST004305
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 PR002721. The data can be accessed directly via it's Project DOI: 10.21228/M8XV8X 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 | ST004305 |
| Study Title | Lactate mitochondrial oxidation drives stemness potential in metastatic breast cancer |
| Study Summary | Metastatic cancer cells, originating from cancer stem cells with metastatic capacity, utilize nutrient flexibility to navigate the challenges of the metastatic cascade. However, the nutrient required to maintain the stemness potentials of metastatic cancer cells remains unclear. Here, we reveal that metastatic breast cancer cells sustain stemness and initiate metastasis upon detachment by taking up and oxidizing lactate. In detached metastasizing breast cancer cells, lactate is incorporated into the tricarboxylic acid cycle, boosting oxidative phosphorylation, and promoting the stemness potentials via α-KG-DNMT3B-mediated SOX2 hypomethylation. Moreover, lactate is taken up and oxidized in mitochondria by the CD147/MCT1/LDHB complex, which correlates with stemness potentials and tumor metastasis in patients with breast cancer. An intracellularly expressed single-chain variable fragment targeting mitochondrial CD147 (mito-CD147 scFv) effectively disrupts the mitochondrial CD147/MCT1/LDHB complex, inhibits lactate-induced stemness potential, depletes circulating breast cancer cells, and reduces metastatic burden, suggesting promising clinical applications in reducing lactate-fueled metastasis. |
| Institute | Fourth Military Medical University |
| Last Name | Zhang |
| First Name | Jia-Jia |
| Address | No. 169 Changle West Road, Xincheng District, Xi'an City, Shaanxi Province |
| 1240568116@qq.com | |
| Phone | 15619276681 |
| Submit Date | 2025-10-11 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | cdf |
| Analysis Type Detail | GC-MS |
| Release Date | 2025-11-07 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002721 |
| Project DOI: | doi: 10.21228/M8XV8X |
| Project Title: | Lactate mitochondrial oxidation drives stemness potential in metastatic breast cancer |
| Project Summary: | Metastatic cancer cells, originating from cancer stem cells with metastatic capacity, utilize nutrient flexibility to navigate the challenges of the metastatic cascade. However, the nutrient required to maintain the stemness potentials of metastatic cancer cells remains unclear. Here, we reveal that metastatic breast cancer cells sustain stemness and initiate metastasis upon detachment by taking up and oxidizing lactate. In detached metastasizing breast cancer cells, lactate is incorporated into the tricarboxylic acid cycle, boosting oxidative phosphorylation, and promoting the stemness potentials via α-KG-DNMT3B-mediated SOX2 hypomethylation. Moreover, lactate is taken up and oxidized in mitochondria by the CD147/MCT1/LDHB complex, which correlates with stemness potentials and tumor metastasis in patients with breast cancer. An intracellularly expressed single-chain variable fragment targeting mitochondrial CD147 (mito-CD147 scFv) effectively disrupts the mitochondrial CD147/MCT1/LDHB complex, inhibits lactate-induced stemness potential, depletes circulating breast cancer cells, and reduces metastatic burden, suggesting promising clinical applications in reducing lactate-fueled metastasis. |
| Institute: | Fourth Military Medical University |
| Last Name: | Zhang |
| First Name: | Jia-Jia |
| Address: | No. 169 Changle West Road, Xincheng District, Xi'an City, Shaanxi Province |
| Email: | 1240568116@qq.com |
| Phone: | +86 156 1927 6681 |
Subject:
| Subject ID: | SU004458 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Genotype Strain: | Wild type |
| Age Or Age Range: | NA |
| Weight Or Weight Range: | NA |
| Height Or Height Range: | NA |
| Gender: | Female |
| Cell Biosource Or Supplier: | ATCC |
| Cell Strain Details: | MCF7 |
| Subject Comments: | NA |
| Cell Primary Immortalized: | No. |
| Cell Passage Number: | 2 |
| Cell Counts: | 1x10e6 |
| Species Group: | human |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Adherent | Sample source |
|---|---|---|---|
| SA505322 | M_Ad_1_1 | Adherent | MCF-7 breast cancer cell line |
| SA505323 | M_Ad_2_1 | Adherent | MCF-7 breast cancer cell line |
| SA505324 | M_Ad_3_1 | Adherent | MCF-7 breast cancer cell line |
| SA505325 | M_Ad_4_1 | Adherent | MCF-7 breast cancer cell line |
| SA505326 | M_Ad_5_1 | Adherent | MCF-7 breast cancer cell line |
| SA505327 | M_Ad_6_1 | Adherent | MCF-7 breast cancer cell line |
| SA505328 | M_AR_1_1 | anoikis-resistant | MCF-7 breast cancer cell line |
| SA505329 | M_AR_2_1 | anoikis-resistant | MCF-7 breast cancer cell line |
| SA505330 | M_AR_3_1 | anoikis-resistant | MCF-7 breast cancer cell line |
| SA505331 | M_AR_4_1 | anoikis-resistant | MCF-7 breast cancer cell line |
| SA505332 | M_AR_5_1 | anoikis-resistant | MCF-7 breast cancer cell line |
| SA505333 | M_AR_6_1 | anoikis-resistant | MCF-7 breast cancer cell line |
| Showing results 1 to 12 of 12 |
Collection:
| Collection ID: | CO004451 |
| Collection Summary: | Supernatants from 1×10e6 MCF7 AR and Ad cells (n=6 biological replicates) were collected and centrifuged at 3000×g for 10 min at 4 ℃ to remove cell debris and the lipid layer. |
| Sample Type: | Cell culture supernatant |
| Collection Method: | Direct collection |
| Collection Frequency: | One time collection |
| Collection Duration: | About 20 min. |
| Volumeoramount Collected: | 1 ml |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR004467 |
| Treatment Summary: | MCF7, cells were cultured in RPMI 1640 medium with 3.15g/L glucose. All cells were routinely supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 2% L-glutamine. Cells surviving for 7 days in glucose-containing medium (3.15g/L) under detached suspension culture conditions in ultra-low attachment plates were considered anoikis-resistant (AR) cells, while cells cultured in uncoated common plates under attached conditions with glucose-containing medium (3.15g/L) served as control adherent (Ad) cells. |
| Treatment: | Adherent (Ad) cells and anoikis-resistant (AR) cells. |
| Cell Storage: | 37℃ |
| Cell Growth Container: | 6-well plate |
| Cell Growth Config: | MCF7 cells were cultured in RPMI 1640 medium with 3.15g/L glucose. All cells were routinely supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 2% L-glutamine. |
| Cell Media: | RPMI 1640 medium |
| Cell Envir Cond: | 37℃, 5% CO2 |
| Cell Harvesting: | Collect the supernatant |
| Cell Pct Confluence: | 90% |
| Cell Media Lastchanged: | Change the medium every three days。 |
Sample Preparation:
| Sampleprep ID: | SP004464 |
| Sampleprep Summary: | The medium samples were thawed on ice-bath and centrifuged for 5 min at 4 °C and 3000g (Microfuge 20R， Beckman Coulter, Inc., Indianapolis, IN, USA) to separate debris or a lipid layer. Each aliquot of 50 μL sample was mixed with 10 μL of internal standard, to which 175 μL of pre-chilled methanol/chloroform (v/v=3/1) was added. After centrifugation at 14,000 g and 4 °C for 20 min (Microfuge 20R，Beckman Coulter, Inc., Indianapolis, IN, USA), each 200 μL of the supernatant was carefully transferred to an autosampler vial (Agilent Technologies, Foster City, CA, USA). The remaining supernatant from each sample was pooled to make quality control samples. All the samples in autosampler vials were evaporated briefly to remove chloroform using a CentriVap vacuum concentrator (Labconco, Kansas City, MO, USA), and further lyophilized with a FreeZone freeze dryer equipped with a stopping tray dryer (Labconco, Kansas City, MO, USA). The sample derivatization and injection were performed by a robotic multipurpose sample MPS2 with dual heads (Gerstel, Muehlheim, Germany). Briefly, the dried sample was derivatized with 50 μL of methoxyamine (20 mg/mL in pyridine) at 30°C for 2 hr, followed by the addition of 50 μL of MSTFA (1% TMCS) containing FAMEs as retention indices at 37.5 °C for another 1 hr using the sample preparation head. In parallel, the derivatized samples were injected with sample injection head after derivatization. |
Combined analysis:
| Analysis ID | AN007162 |
|---|---|
| Chromatography ID | CH005441 |
| MS ID | MS006857 |
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 7890B |
| Column | Restek Rtx-5Sil MS (30m x 0.25mm, 0.25um) |
| MS Type | EI |
| MS instrument type | TOF |
| MS instrument name | Leco Pegasus HT TOF |
| Ion Mode | POSITIVE |
| Units | z-score |
Chromatography:
| Chromatography ID: | CH005441 |
| Chromatography Summary: | A time-of-flight mass spectrometry (GC-TOF/MS) system (Pegasus HT, Leco Corp., St. Joseph, MO,USA) with an Agilent 7890B gas chromatography and a Gerstel multipurpose sample MPS2 with dual heads (Gerstel, Muehlheim, Germany). A Rxi-5 ms capillary column (30m×250μm i.d., 0.25-μm film thickness; Restek corporation, Bellefonte, PA, USA) was used for separation. Helium was used as the carrier gas at a constant flow rate of 1.0 mL/min. The temperature of injection and transfer interface were both set to 270 °C. The source temperature was 220 °C. The measurements were made using electron impact ionization (70 eV) in the full scan mode (m/z 50-500). |
| Instrument Name: | Agilent 7890B |
| Column Name: | Restek Rtx-5Sil MS (30m x 0.25mm, 0.25um) |
| Column Temperature: | 270 °C |
| Flow Gradient: | NA |
| Flow Rate: | 1.0 mL/min |
| Solvent A: | NA |
| Solvent B: | NA |
| Chromatography Type: | GC |
MS:
| MS ID: | MS006857 |
| Analysis ID: | AN007162 |
| Instrument Name: | Leco Pegasus HT TOF |
| Instrument Type: | TOF |
| MS Type: | EI |
| MS Comments: | Metabolite Annotation Metabolite annotation was performed by comparing the retention indices and mass spectral data withthose previously generated from reference standards of known structures present in JiaLib metabolite database using ChromaTOF software . The current JiaLib comprises over 1,200 mammalian metabolites with15-year accumulation and is one of the most comprehensive metabolite libraries in the world. The referencechemicals present in JiaLib were commercially purchased from Sigma-Aldrich (St. Louis, MO, USA), Santa Cruz (Dallas, TX, USA), Nu-Chek Prep (Elysian, MN, USA), and synthesized in the laboratory.Commercial libraries such as NIST library 2014 and LECO/Fiehn Metabolomics Library for GC-TOF/MS are NOT recommended for precise metabolite annotation for the following two reasons: 1) neither instrument vendor’s software nor commercially available software can annotate metabolites using both retention indices and mass spectral data; and 2) both NIST and Fiehn libraries contain a large portion of reference materials that are intended for chemical annotation in food, plant, drug, and synthesized chemistry. Data Analysis and Software The raw data generated by GC-TOF/MS were processed using ChromaTOF (v4.71, Leco Corp., St.Joseph, MO,USA) for automated baseline denosing and smoothing, peak picking, deconvolution and peak alignment. Compound identification was performed by comparing both MS similarity and FAMEs retention index distance with the referenced standards in one of the most extensive metabolite database – JiaLib in the world. The self-developed platform iMAP（v1.0, Metabo-Profile, Shanghai, China）was used for subsequent statistical analyses, including PCA, OPLS-DA, univariate analysis and pathway analysis, et al. Statistics Our proprietary software can perform a collection of data processing, interpretation, and visualization. For many metabolomics studies, two types of statistical analysis are extensively performed: 1) multivariate statistical analyses such as principal component analysis (PCA), partial least square discriminant analysis (PLS-DA), orthogonal partial least square discriminant analysis (OPLS-DA), random forest, support vector machine learning and so on; 2) univariate statistical analyses including student t-test, Mann-Whitney-Wilcoxon (U-test), ANOVA, correlation analysis, etc. Statistical algorithms are adapted from the widely used statistical analysis software packages in R studio (http://cran.r-project.org/). The optimal choice of statistical methods is often driven by the data and the project goals. |
| Ion Mode: | POSITIVE |
