Production of hyperpolarized [1,4-¹³C₂]malate from [1,4-¹³C₂]fumarate is a marker of cell necrosis and treatment response in tumors

• Published in: Proceedings of the National Academy of Sciences - PNAS 2009-11, Vol.106 (47), p.19801-19806
• Main Authors: Gallagher, Ferdia A, Kettunen, Mikko I, Hu, De-En, Jensen, Pernille R, Zandt, René in 't, Karlsson, Magnus, Gisselsson, Anna, Nelson, Sarah K, Witney, Timothy H, Bohndiek, Sarah E, Hansson, Georg, Peitersen, Torben, Lerche, Mathilde H, Brindle, Kevin M
• Format: Article
• Language: English
• Subjects: Animals; Antineoplastic Agents, Phytogenic - therapeutic use; Antineoplastics; Apoptosis; Biological Sciences; Biomarkers, Tumor - chemistry; Biomarkers, Tumor - metabolism; Cancer; Carbon Isotopes - chemistry; Carbon Isotopes - metabolism; Cell death; Cells; Etoposide - therapeutic use; Experiments; Female; Fumarate Hydratase - metabolism; Fumarates - chemistry; Fumarates - metabolism; Imaging; Lymphoma; Lymphoma - metabolism; Lymphoma - pathology; Magnetic resonance imaging; Malates - chemistry; Malates - metabolism; Mice; Mice, Inbred C57BL; Molecules; Necrosis; Necrosis - metabolism; Necrosis - pathology; Neoplasm Transplantation; Neoplasms - drug therapy; Neoplasms - metabolism; Neoplasms - pathology; NMR; Nuclear magnetic resonance; Nuclear Magnetic Resonance, Biomolecular; Tissue Extracts - metabolism; Treatment Outcome; Tumors
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0911447106

Dynamic nuclear polarization of ¹³C-labeled cell substrates has been shown to massively increase their sensitivity to detection in NMR experiments. The sensitivity gain is sufficiently large that if these polarized molecules are injected intravenously, their spatial distribution and subsequent conversion into other cell metabolites can be imaged. We have used this method to image the conversion of fumarate to malate in a murine lymphoma tumor in vivo after i.v. injection of hyperpolarized [1,4-¹³C₂]fumarate. In isolated lymphoma cells, the rate of labeled malate production was unaffected by coadministration of succinate, which competes with fumarate for transport into the cell. There was, however, a correlation with the percentage of cells that had lost plasma membrane integrity, suggesting that the production of labeled malate from fumarate is a sensitive marker of cellular necrosis. Twenty-four hours after treating implanted lymphoma tumors with etoposide, at which point there were significant levels of tumor cell necrosis, there was a 2.4-fold increase in hyperpolarized [1,4-¹³C₂]malate production compared with the untreated tumors. Therefore, the formation of hyperpolarized ¹³C-labeled malate from [1,4-¹³C₂]fumarate appears to be a sensitive marker of tumor cell death in vivo and could be used to detect the early response of tumors to treatment. Given that fumarate is an endogenous molecule, this technique has the potential to be used clinically.