Real-time measurement of hyperpolarized lactate production and efflux as a biomarker of tumor aggressiveness in an MR compatible 3D cell culture bioreactor Journal Article
| Authors: | Sriram, R.; Van Criekinge, M.; Hansen, A.; Wang, Z. J.; Vigneron, D. B.; Wilson, D. M.; Keshari, K. R.; Kurhanewicz, J. |
| Article Title: | Real-time measurement of hyperpolarized lactate production and efflux as a biomarker of tumor aggressiveness in an MR compatible 3D cell culture bioreactor |
| Abstract: | We have developed a 3D cell/tissue culture bioreactor compatible with hyperpolarized (HP) 13C MR and interrogated HP [1-13C]lactate production and efflux in human renal cell carcinoma (RCC) cells. This platform is capable of resolving intracellular and extracellular HP lactate pools, allowing the kinetic measurement of lactate production and efflux in the context of cancer aggressiveness and response to therapy. HP 13C MR studies were performed on three immortalized human renal cell lines: HK2, a normal renal proximal tubule cell line from which a majority of RCCs arise, UMRC6, a cell line derived from a localized RCC, and UOK262, an aggressive and metastatic RCC. The intra- (Lac<inf>in</inf>) and extracellular (Lac<inf>ex</inf>) HP lactate signals were robustly resolved in dynamic 13C spectra of the cell lines due to a very small but reproducible chemical shift difference (0.031 ± 0.0005 ppm). Following HP [1-13C]pyruvate delivery, the ratio of HP Lac<inf>in</inf>/Lac<inf>ex</inf> was significantly lower for UOK262 cells compared with both UMRC6 and HK2 cells due to a significant (p < 0.05) increase in the Lac<inf>ex</inf> pool size. Lac<inf>in</inf>/Lac<inf>ex</inf> correlated with the MCT4 mRNA expression of the cell lines, and inhibition of MCT4 transport using DIDS resulted in a significant reduction in the HP Lac<inf>ex</inf> pool size. The extension of these studies to living patient-derived RCC tissue slices using HP [1,2-13C<inf>2</inf>]pyruvate demonstrated a similarly split lactate doublet with a high Lac<inf>ex</inf> pool fraction; in contrast, only a single NMR resonance is noted for HP [5-13C]glutamate, consistent with intracellular localization. These studies support the importance of lactate efflux as a biomarker of cancer aggressiveness and metastatic potential, and the utility of the MR compatible 3D cell/tissue culture bioreactor to study not only cellular metabolism but also transport. Additionally, this platform offers a sophisticated way to follow therapeutic interventions and screen novel therapies that target lactate export. © 2015 John Wiley & Sons, Ltd. |
| Keywords: | human tissue; human cell; biological marker; cytology; metastasis; gene expression; enzyme activity; renal cell carcinoma; carcinogenesis; correlation analysis; cell culture; messenger rna; magnetic resonance; cellular distribution; glutamic acid; cell metabolism; nuclear magnetic resonance; diseases; bioreactor; bioreactors; proton nuclear magnetic resonance; carbon nuclear magnetic resonance; lactic acid; cells; pyruvic acid; pyruvate; lactate; aerobic glycolysis; hyperpolarized; hyperpolarization; chemical shift; human; priority journal; article; mobile security; cancer aggressiveness; dynamic nuclear polarization (dnp); hyperpolarized 13c magnetic resonance (hp 13c mr); lactate efflux; renal cell carcinoma (rcc); bioconversion; lakes; dynamic nuclear polarization; pyruvates; monocarboxylate transporter 4; renal cell carcinoma cell line |
| Journal Title: | NMR in Biomedicine |
| Volume: | 28 |
| Issue: | 9 |
| ISSN: | 0952-3480 |
| Publisher: | John Wiley & Sons |
| Date Published: | 2015-09-01 |
| Start Page: | 1141 |
| End Page: | 1149 |
| Language: | English |
| DOI: | 10.1002/nbm.3354 |
| PROVIDER: | scopus |
| PMCID: | PMC4537810 |
| PUBMED: | 26202449 |
| DOI/URL: | |
| Notes: | Export Date: 2 September 2015 -- Source: Scopus |
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