Summary of Study ST001731
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 PR001108. The data can be accessed directly via it's Project DOI: 10.21228/M8GM5B 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 | ST001731 |
| Study Title | Mitochondrial ATP fuels ABC transporter-mediated drug efflux in cancer chemoresistance (part-II) |
| Study Summary | Chemotherapy remains the standard of care for most cancers worldwide, however development of chemoresistance due to the presence of the drug-effluxing ABC transporters remains a significant problem. The development of safe and effective means to overcome chemoresistance is critical for achieving durable remissions in many cancer patients. We have investigated the energetic demands of ABC transporters in the context of the metabolic adaptations of chemoresistant cancer cells. Here we show that ABC transporters use mitochondrial-derived ATP as a source of energy to efflux drugs out of cancer cells. We further demonstrate that the loss of MCJ (DnaJC15), an endogenous negative regulator of mitochondrial respiration, in chemoresistant cancer cells boosts their ability to produce ATP from mitochondria and fuel ABC transporters. We have developed novel MCJ mimetics that can attenuate mitochondrial respiration and safely overcome chemoresistance in vitro and in vivo. Administration of MCJ mimetics in combination with standard chemotherapeutic drugs could therefore become an new strategy for treatment of multiple cancers. |
| Institute | University of Colorado Denver |
| Department | Biochemistry and Molecular Genetics |
| Laboratory | Angelo D'Alessandro |
| Last Name | Culp-Hill |
| First Name | Rachel |
| Address | 12801 E 17th Ave L18-9403D, Aurora, Colorado, 80045, USA |
| rachel.hill@cuanschutz.edu | |
| Phone | 303-724-5798 |
| Submit Date | 2021-03-18 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2021-03-31 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001108 |
| Project DOI: | doi: 10.21228/M8GM5B |
| Project Title: | Mitochondrial ATP fuels ABC transporter-mediated drug efflux in cancer chemoresistance |
| Project Summary: | Chemotherapy remains the standard of care for most cancers worldwide, however development of chemoresistance due to the presence of the drug-effluxing ABC transporters remains a significant problem. The development of safe and effective means to overcome chemoresistance is critical for achieving durable remissions in many cancer patients. We have investigated the energetic demands of ABC transporters in the context of the metabolic adaptations of chemoresistant cancer cells. Here we show that ABC transporters use mitochondrial-derived ATP as a source of energy to efflux drugs out of cancer cells. We further demonstrate that the loss of MCJ (DnaJC15), an endogenous negative regulator of mitochondrial respiration, in chemoresistant cancer cells boosts their ability to produce ATP from mitochondria and fuel ABC transporters. We have developed novel MCJ mimetics that can attenuate mitochondrial respiration and safely overcome chemoresistance in vitro and in vivo. Administration of MCJ mimetics in combination with standard chemotherapeutic drugs could therefore become an new strategy for treatment of multiple cancers. |
| Institute: | University of Colorado Denver |
| Department: | Biochemistry and Molecular Genetics |
| Laboratory: | Angelo D'Alessandro |
| Last Name: | Culp-Hill |
| First Name: | Rachel |
| Address: | 12801 E 17th Ave L18-9403D, Aurora, Colorado, 80045, USA |
| Email: | rachel.hill@cuanschutz.edu |
| Phone: | 303-724-5798 |
Subject:
| Subject ID: | SU001808 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Factor |
|---|---|---|
| SA162629 | control1 | Control |
| SA162630 | control4 | Control |
| SA162631 | control3 | Control |
| SA162632 | control2 | Control |
| SA162633 | siMCJ4 | siMCJkd |
| SA162634 | siMCJ1 | siMCJkd |
| SA162635 | siMCJ2 | siMCJkd |
| SA162636 | siMCJ3 | siMCJkd |
| Showing results 1 to 8 of 8 |
Collection:
| Collection ID: | CO001801 |
| Collection Summary: | MCF7 cells were purchased from American Type Culture Collection (ATCC). All cancer cell lines were maintained in RPMI 1640 (Sigma R8758) containing glucose (2 mg/ml) and glutamine (0.6 mg/ml) but no pyruvate and was supplemented with 5% Fetal calf serum. |
| Sample Type: | Cultured cells |
Treatment:
| Treatment ID: | TR001821 |
| Treatment Summary: | MCF7 cell transfections were performed using siPORT NeoFX Transfection Agent (ThermoFisher Scientific) following the recommended protocol for an oligonucleotide siRNA based on the previously described MCJ targeting sequence. |
Sample Preparation:
| Sampleprep ID: | SP001814 |
| Sampleprep Summary: | All cancer cell lines were maintained in RPMI 1640 (Sigma R8758) containing glucose (2 mg/ml) and glutamine (0.6 mg/ml) but no pyruvate and was supplemented with 5% Fetal calf serum. Metabolic profile comparisons of cells were performed on equal cell numbers. Cells were cultured under normal conditions, detached using trypsin-EDTA (0.05 %), counted, normalized to 0.5 x 10e6 in each sample, and cell pellets were snap frozen in liquid nitrogen prior to analysis. Briefly, 2 x 10e6 cells and 20μL cell media were extracted in 1mL and 980μL cold lysis and extraction buffer (methanol : acetonitrile : water, 5:3:2), respectively, and pellets were discarded. |
Combined analysis:
| Analysis ID | AN002817 | AN002818 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| Column | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Relative Abundance | Relative Abundance |
Chromatography:
| Chromatography ID: | CH002082 |
| Chromatography Summary: | Positive Mode 10 μL of water and methanol soluble fractions were run through a Kinetex C18 1.7 μm, 100 x 2.1 mm (Phenomenex) reversed phase column (Positive ion mode - phase A: water, 0.1 % formic acid; B: acetonitrile, 0.1 % formic acid) via an ultra-high performance chromatographic system (UHPLC - Vanquish, Thermo Fisher). UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in positive polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) |
| Solvent A: | 100% water; 0.1 % formic acid |
| Solvent B: | 100% acetonitrile, 0.1 % formic acid |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH002083 |
| Chromatography Summary: | Negative Mode 10 μL of water and methanol soluble fractions were run through a Kinetex C18 1.7 μm, 100 x 2.1 mm (Phenomenex) reversed phase column (Negative ion mode – phase A: 1 mM NH4Ac 95:5 water : acetonitrile; phase B: 1 mM NH4Ac 95:5 acetonitrile : water) via an ultra-high performance chromatographic system (UHPLC - Vanquish, Thermo Fisher). UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in negative polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) |
| Solvent A: | 95% water/5% acetonitrile; 1 mM ammonium acetate |
| Solvent B: | 95% acetonitrile/5% water; 1 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002611 |
| Analysis ID: | AN002817 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in positive polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described. |
| Ion Mode: | POSITIVE |
| MS ID: | MS002612 |
| Analysis ID: | AN002818 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in negative polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described. |
| Ion Mode: | NEGATIVE |
