1H and 31P Magnetic Resonance Spectroscopic Metabolomic Imaging: Assessing Mitogen-Activated Protein Kinase Inhibition in Melanoma
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents and MEK Inhibitor
2.2. Cell Lines and Cell Culture Conditions
2.3. Extracellular Glucose and Lactate Measurements
2.4. Mitochondrial Stress and Cell Energy Phenotype Assays
2.5. In Vitro Measurement of Intracellular Metabolites by High-Resolution 1H-MRS
2.6. Mouse Preparation for Proton (1H) and Phosphorus (31P) MRS Studies
2.7. Experiments Involving In Vivo 1H and 31P MRS
2.8. Measurement of Tumor Volume
2.9. Statistical Analysis
3. Results
3.1. Trametinib’s Impact on Glucose Consumption and Lactate Production in Melanoma In Vitro
3.2. Oxygen Consumption and Extracellular Acidification Rates In Vitro
Group | Mean ± SD (n = 8) | p-Value | |
---|---|---|---|
Control | Trametinib | ||
Oxygen consumption rate (pmol/min/1000 cells) | |||
WM3918 | 7.09 ± 0.99 | 6.49 ± 1.01 | 0.13 |
WM983BR | 2.79 ± 0.50 | 3.50 ± 0.31 | 0.007 |
WM983B | 4.63 ± 0.41 | 5.06 ± 0.48 | 0.014 |
DB-1 | 7.25 ± 0.36 | 5.05 ± 0.46 | <0.001 |
Extracellular acidification rate (pmol/min/1000 cells) | |||
WM3918 | 2.31 ± 0.24 | 2.04 ± 0.26 | 0.011 |
WM983BR | 3.10 ± 0.46 | 4.01 ± 0.54 | 0.015 |
WM983B | 2.31 ± 0.22 | 1.78 ± 0.21 | 0.001 |
DB-1 | 2.95 ± 0.29 | 2.38 ± 0.53 | 0.001 |
OCR/ECAR ratio—cell energy phenotype | |||
WM3918 | 3.08 ± 0.37 | 3.18 ± 0.29 | 0.46 |
WM983BR | 0.90 ± 0.12 | 0.88 ± 0.08 | 0.71 |
WM983B | 2.02 ± 0.22 | 2.89 ± 0.55 | <0.001 |
DB-1 | 2.48 ± 0.28 | 2.17 ± 0.33 | 0.01 |
3.3. In Vitro 1H MRS to Test the Metabolic Response to Trametinib in Isolated Melanoma Cells
3.4. In Vivo 1H- and 31P- MRS of Melanoma Xenografts in Mice
3.5. Assessment of Tumor Growth Following Trametinib Treatment
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gupta, P.K.; Orlovskiy, S.; Arias-Mendoza, F.; Nelson, D.S.; Nath, K. 1H and 31P Magnetic Resonance Spectroscopic Metabolomic Imaging: Assessing Mitogen-Activated Protein Kinase Inhibition in Melanoma. Cells 2024, 13, 1220. https://doi.org/10.3390/cells13141220
Gupta PK, Orlovskiy S, Arias-Mendoza F, Nelson DS, Nath K. 1H and 31P Magnetic Resonance Spectroscopic Metabolomic Imaging: Assessing Mitogen-Activated Protein Kinase Inhibition in Melanoma. Cells. 2024; 13(14):1220. https://doi.org/10.3390/cells13141220
Chicago/Turabian StyleGupta, Pradeep Kumar, Stepan Orlovskiy, Fernando Arias-Mendoza, David S. Nelson, and Kavindra Nath. 2024. "1H and 31P Magnetic Resonance Spectroscopic Metabolomic Imaging: Assessing Mitogen-Activated Protein Kinase Inhibition in Melanoma" Cells 13, no. 14: 1220. https://doi.org/10.3390/cells13141220