Modulation of choline kinase activity in human cancer cells observed by dynamic <sup>31</sup>P NMR

Gabellieri, Cristina; Beloueche‐Babari, Mounia; Jamin, Yann; Payne, Geoffrey S.; Leach, Martin O.; Eykyn, Thomas R.

doi:10.1002/nbm.1361

Cited by 16 publications

(9 citation statements)

References 32 publications

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“…Our MS imaging findings confirm the importance of PC in malignant tumor growth (23). Altered expression and activity levels of phosphatidylcholine metabolism-related enzymes, such as choline kinase (5, 8, 34), phospholipases C and D (42) and specific choline transporters (43) may account for the higher MS imaging-detected PC intensity in viable regions of the more aggressive tumor model. A recent studies in prostate tumor models detected increased tCho and PC concentrations in hypoxic tumor regions, and showed that hypoxia inducible factor-1 (HIF-1) signaling increased choline kinase expression, and hence PC production, under hypoxic conditions (3).…”

Section: Discussionmentioning

confidence: 99%

“…Increases in activity and/or expression of several enzymes in choline metabolism, such as choline transporters (7), choline kinase (8), and phospholipases C (9) and D (10, 11) are responsible for the elevated PC and tCho concentrations in cancers, and provide potential therapeutic targets. Choline kinase has already been developed as a target for treatment (5, 12) and a predictive marker for cancer prognosis (5).…”

Section: Introductionmentioning

confidence: 99%

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Multimodal Mass Spectrometric Imaging of Small Molecules Reveals Distinct Spatio-Molecular Signatures in Differentially Metastatic Breast Tumor Models

et al. 2010

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Phosphocholine (PC) and total choline (tCho) are increased in malignant breast tumors. In this study, we combined magnetic resonance spectroscopic imaging (MRSI), mass spectrometry (MS) imaging, and pathological assessment of corresponding tumor sections, to investigate the localization of choline metabolites and cations in viable versus necrotic tumor regions in the nonmetastatic MCF-7 and the highly metastatic MDA-MB-231 breast cancer xenograft models. In vivo 3-dimensional MRSI demonstrated that high tCho levels, consisting of free choline (Cho), PC, and glycerophosphocholine (GPC), displayed a heterogeneous spatial distribution in the tumor. MS imaging performed on tumor sections detected the spatial distributions of individual PC, Cho, and GPC, as well as sodium (Na+) and potassium (K+), among many others. PC and Cho intensity were increased in viable compared to necrotic regions of MDA-MB-231 tumors, but relatively homogeneously distributed in MCF-7 tumors. Such behavior may be related to the role of PC and PC-related enzymes, such as choline kinase, choline transporters and others in malignant tumor growth. Na+ and K+ colocalized in the necrotic tumor areas of MDA-MB-231 tumors, whereas in MCF-7 tumors, Na+ was detected in necrotic and K+ in viable tumor regions. This may be attributed to differential Na+/K+ pump functions and K+ channel expressions. Principal component analysis of the MS imaging data clearly identified different tumor microenvironmental regions by their distinct molecular signatures. This molecular information allowed us to differentiate between distinct tumor regions and tumor types, which may, in the future, prove clinically useful in the pathological assessment of breast cancers.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Multimodal Mass Spectrometric Imaging of Small Molecules Reveals Distinct Spatio-Molecular Signatures in Differentially Metastatic Breast Tumor Models

et al. 2010

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“…Experiments were performed to assay the total cellular activity of choline kinase as previously described [31], [32]. In brief, cell pellets (2×10 7 ) were resuspended in 540 µL lysis buffer containing 100 mM Tris-HCl (pH 8.0), 10 mM DTT and 1 mM EDTA in D 2 O. Lysate was homogenized on ice by repeated passage through fine-tipped needle (27G 1/2) and further disrupted by sonication 10×1s at 20 kHz.…”

Section: Methodsmentioning

confidence: 99%

HDAC Inhibition Induces Increased Choline Uptake and Elevated Phosphocholine Levels in MCF7 Breast Cancer Cells

et al. 2013

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Histone deacetylase (HDAC) inhibitors have emerged as effective antineoplastic agents in the clinic. Studies from our lab and others have reported that magnetic resonance spectroscopy (MRS)-detectable phosphocholine (PC) is elevated following SAHA treatment, providing a potential noninvasive biomarker of response. Typically, elevated PC is associated with cancer while a decrease in PC accompanies response to antineoplastic treatment. The goal of this study was therefore to elucidate the underlying biochemical mechanism by which HDAC inhibition leads to elevated PC. We investigated the effect of SAHA on MCF-7 breast cancer cells using 13C MRS to monitor [1,2-13C] choline uptake and phosphorylation to PC. We found that PC synthesis was significantly higher in treated cells, representing 154±19% of control. This was within standard deviation of the increase in total PC levels detected by 31P MRS (129±7% of control). Furthermore, cellular choline kinase activity was elevated (177±31%), while cytidylyltransferase activity was unchanged. Expression of the intermediate-affinity choline transporter SLC44A1 and choline kinase α increased (144% and 161%, respectively) relative to control, as determined by mRNA microarray analysis with protein-level confirmation by Western blotting. Taken together, our findings indicate that the increase in PC levels following SAHA treatment results from its elevated synthesis. Additionally, the concentration of glycerophosphocholine (GPC) increased significantly with treatment to 210±45%. This is likely due to the upregulated expression of several phospholipase A2 (PLA2) isoforms, resulting in increased PLA2 activity (162±18%) in SAHA-treated cells. Importantly, the levels of total choline (tCho)-containing metabolites, comprised of choline, PC and GPC, are readily detectable clinically using 1H MRS. Our findings thus provide an important step in validating clinically translatable non-invasive imaging methods for follow-up diagnostics of HDAC inhibitor treatment.

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“…CHK activity was assayed using an MRS-based method as described [54], with modifications. Briefly, about 25 million cells were collected by trypsinization and washed in PBS.…”

Section: Chk Activity Assaymentioning

confidence: 99%

De novo phosphatidylcholine synthesis is required for autophagosome membrane formation and maintenance during autophagy

et al. 2019

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2020) Denovo phosphatidylcholine synthesis is required for autophagosome membrane formation and maintenance during autophagy, ABSTRACT Macroautophagy/autophagy can enable cancer cells to withstand cellular stress and maintain bioenergetic homeostasis by sequestering cellular components into newly formed double-membrane vesicles destined for lysosomal degradation, potentially affecting the efficacy of anti-cancer treatments. Using 13 C-labeled choline and 13 C-magnetic resonance spectroscopy and western blotting, we show increased de novo choline phospholipid (ChoPL) production and activation of PCYT1A (phosphate cytidylyltransferase 1, choline, alpha), the rate-limiting enzyme of phosphatidylcholine (PtdCho) synthesis, during autophagy. We also discovered that the loss of PCYT1A activity results in compromised autophagosome formation and maintenance in autophagic cells. Direct tracing of ChoPLs with fluorescence and immunogold labeling imaging revealed the incorporation of newly synthesized ChoPLs into autophagosomal membranes, endoplasmic reticulum (ER) and mitochondria during anticancer drug-induced autophagy. Significant increase in the colocalization of fluorescence signals from the newly synthesized ChoPLs and mCherry-MAP1LC3/LC3 (microtubule-associated protein 1 light chain 3) was also found on autophagosomes accumulating in cells treated with autophagy-modulating compounds. Interestingly, cells undergoing active autophagy had an altered ChoPL profile, with longer and more unsaturated fatty acid/ alcohol chains detected. Our data suggest that de novo synthesis may be required to increase autophagosomal ChoPL content and alter its composition, together with replacing phospholipids consumed from other organelles during autophagosome formation and turnover. This addiction to de novo ChoPL synthesis and the critical role of PCYT1A may lead to development of agents targeting autophagyinduced drug resistance. In addition, fluorescence imaging of choline phospholipids could provide a useful way to visualize autophagosomes in cells and tissues.

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Modulation of choline kinase activity in human cancer cells observed by dynamic ³¹P NMR

Cited by 16 publications

References 32 publications

Multimodal Mass Spectrometric Imaging of Small Molecules Reveals Distinct Spatio-Molecular Signatures in Differentially Metastatic Breast Tumor Models

Multimodal Mass Spectrometric Imaging of Small Molecules Reveals Distinct Spatio-Molecular Signatures in Differentially Metastatic Breast Tumor Models

HDAC Inhibition Induces Increased Choline Uptake and Elevated Phosphocholine Levels in MCF7 Breast Cancer Cells

De novo phosphatidylcholine synthesis is required for autophagosome membrane formation and maintenance during autophagy

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Modulation of choline kinase activity in human cancer cells observed by dynamic 31P NMR

Cited by 16 publications

References 32 publications

Multimodal Mass Spectrometric Imaging of Small Molecules Reveals Distinct Spatio-Molecular Signatures in Differentially Metastatic Breast Tumor Models

Multimodal Mass Spectrometric Imaging of Small Molecules Reveals Distinct Spatio-Molecular Signatures in Differentially Metastatic Breast Tumor Models

HDAC Inhibition Induces Increased Choline Uptake and Elevated Phosphocholine Levels in MCF7 Breast Cancer Cells

De novo phosphatidylcholine synthesis is required for autophagosome membrane formation and maintenance during autophagy

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Modulation of choline kinase activity in human cancer cells observed by dynamic ³¹P NMR