Effect of diclofenac on SLC16A3/MCT4 by the Caco-2 cell line

Sasaki, Shotaro; Futagi, Yuya; Ideno, Masaya; Kobayashi, Masaki; Narumi, Katsuya; Furugen, Ayako; Iseki, Ken

doi:10.1016/j.dmpk.2016.03.004

Cited by 42 publications

(30 citation statements)

References 26 publications

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“…Another potential way to meet this need is the use of NSAIDs, which have been shown to impair lactate efflux and glucose metabolism (Emoto et al, 2002;Gottfried et al, 2013). In accordance with our studies, Sasaki et al (2016) investigated the impact of diclofenac on MCT activity in oocytes expressing either MCT1 or MCT4. Here, we compared the impact of diclofenac, lumiracoxib, ketoprofen, and aspirin in Xenopus oocytes expressing either MCT1 or MCT4 and calculated their IC 50 values.…”

Section: Discussionsupporting

confidence: 55%

Restricting Glycolysis Preserves T Cell Effector Functions and Augments Checkpoint Therapy

et al. 2019

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Highlights d Glycolytic index in melanoma negatively correlates with response to anti-PD1 therapy d Blocking lactate transport or knock out of glycolytic genes improves checkpoint therapy d Diclofenac blocks the lactate transporters MCT1 and MCT4 in a COX-independent manner d Inhibition of glycolysis by MCT blockade does not impede T cell function SUMMARY Tumor-derived lactic acid inhibits T and natural killer (NK) cell function and, thereby, tumor immunosurveillance.Here, we report that melanoma patients with high expression of glycolysis-related genes show a worse progression free survival upon anti-PD1 treatment. The non-steroidal anti-inflammatory drug (NSAID) diclofenac lowers lactate secretion of tumor cells and improves anti-PD1-induced T cell killing in vitro. Surprisingly, diclofenac, but not other NSAIDs, turns out to be a potent inhibitor of the lactate transporters monocarboxylate transporter 1 and 4 and diminishes lactate efflux. Notably, T cell activation, viability, and effector functions are preserved under diclofenac treatment and in a low glucose environment in vitro. Diclofenac, but not aspirin, delays tumor growth and improves the efficacy of checkpoint therapy in vivo. Moreover, genetic suppression of glycolysis in tumor cells strongly improves checkpoint therapy. These findings support the rationale for targeting glycolysis in patients with high glycolytic tumors together with checkpoint inhibitors in clinical trials.

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Section: Discussionsupporting

confidence: 55%

Restricting Glycolysis Preserves T Cell Effector Functions and Augments Checkpoint Therapy

et al. 2019

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“…Recently, the impact of diclofenac on lactate transport was confirmed in MCT-expressing oocytes (131). In a glioma model, diclofenac lowered lactate levels, decreased tumor growth, and tumor-infiltrating dendritic cells regained their capacity to produce IL-12.…”

Section: Targeting Tumor Metabolismmentioning

confidence: 96%

Metabolic Hallmarks of Tumor and Immune Cells in the Tumor Microenvironment

et al. 2017

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Cytotoxic T lymphocytes and NK cells play an important role in eliminating malignant tumor cells and the number and activity of tumor-infiltrating T cells represent a good marker for tumor prognosis. Based on these findings, immunotherapy, e.g., checkpoint blockade, has received considerable attention during the last couple of years. However, for the majority of patients, immune control of their tumors is gray theory as malignant cells use effective mechanisms to outsmart the immune system. Increasing evidence suggests that changes in tumor metabolism not only ensure an effective energy supply and generation of building blocks for tumor growth but also contribute to inhibition of the antitumor response. Immunosuppression in the tumor microenvironment is often based on the mutual metabolic requirements of immune cells and tumor cells. Cytotoxic T and NK cell activation leads to an increased demand for glucose and amino acids, a well-known feature shown by tumor cells. These close metabolic interdependencies result in metabolic competition, limiting the proliferation, and effector functions of tumor-specific immune cells. Moreover, not only nutrient restriction but also tumor-driven shifts in metabolite abundance and accumulation of metabolic waste products (e.g., lactate) lead to local immunosuppression, thereby facilitating tumor progression and metastasis. In this review, we describe the metabolic interplay between immune cells and tumor cells and discuss tumor cell metabolism as a target structure for cancer therapy. Metabolic (re)education of tumor cells is not only an approach to kill tumor cells directly but could overcome metabolic immunosuppression in the tumor microenvironment and thereby facilitate immunotherapy.

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“…2A-B). Moreover, diclofenac, a structurallyunrelated MCT1 & MCT4 blocker 32 , also abrogated the influx of lactate whereas AZD3965 33 , a structurally-unrelated blocker of MCT1 & MCT2 (but not of MCT4), had no effect (Figs. 2C-D).…”

Section: Mct4 Mediates Monocarboxylate Transport In Mda-mb-231 Cellsmentioning

confidence: 98%

MCT4 is a high affinity transporter capable of exporting lactate in high-lactate environments

Contreras-Baeza

Sandoval

Alarcon

et al. 2019

Preprint

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MCT4 is an H + -coupled transporter expressed in metastatic cancer cells, macrophages, and other highly glycolytic cells, where it extrudes excess lactate generated by the Warburg phenomenon or by hypoxia. Intriguingly, its reported K m for lactate, obtained with pH-sensitive probes, is more than an order of magnitude higher than physiological lactate. Here we examined MCT4-rich MDA-MB-231 cells using the FRET sensor Laconic and found a median K m for lactate uptake of only 1.7 mM, while parallel estimation in the same cells with a pH probe gave a K m of 27 mM. The median K m of MCT4 for lactate was 0.7 mM in MCT4-expressing HEK293 cells and 1.2 mM in human macrophages, suggesting that high substrate affinity is a robust property of the transporter. Probed with the FRET sensor Pyronic, MCT4 showed a K m for pyruvate of only 4.2 mM in MDA-MB-231 cells, as opposed to > 150 mM reported previously. We conclude that prior estimates of MCT4 affinity based on pH probes were severely biased by the confounding action of pH regulatory mechanisms. Numerical simulation showed that MCT4, but not MCT1 or MCT2, endows cells with the capability of lactate extrusion in high lactate environments. The revised kinetic properties and novel transport assays may help in developing small-molecule MCT4 blockers for research and therapy. METHODSStandard reagents and inhibitors were acquired from Sigma or Merck. Plasmids encoding the sensors Laconic 18 and Pyronic 19 are available from Addgene (www.addgene.org). Ad Laconic and Ad Pyronic (serotype 5) were custom made by Vector Biolabs. Cell cultureMDA-MB-231 cells were acquired from the American Type Culture Collection (ATCC) and cultured at 37 o C without CO 2 in Leibovitz medium (ThermoFisher). Cultures were transfected at 60% confluence using Lipofectamine 3000 (ThermoFisher) or alternatively, exposed to 5 x 10 6 PFU of Ad Laconic or Ad Pyronic and studied after 24-72 h. The generation of the MDA-LAC cell line will be described elsewhere (Mito Toxy reporter; Contreras et al, submitted doi: https://doi.org/10.1101/583096). HEK293 cells were acquired from the ATCC and cultured at 37 o C in 95% air/5% CO 2 in DMEM/F12 supplemented with 10% fetal bovine serum. Cultures were transfected at 60% confluence using Lipofectamine 3000 (ThermoFisher) and studied after 24-72 h. To obtain macrophages, blood was collected by venepuncture from ten healthy male volunteers. Age of donors ranged from 25 to 45 years. Ethical guidelines stipulated by the Declaration of Helsinki principles were adhered to. Approval was obtained from the Medical Ethical Committee of the Faculty of Medicine, Universidad Austral de Chile. All donors were informed about the nature of the studies and gave their written consent to participate. Samples were treated anonymously. Monocytes were isolated from whole blood treated with 3.8% sodium citrate, by Percoll TM density gradient centrifugation (GE Healthcare, Sweden). Macrophage differentiation of human monocytes was induced by treatment with 25 nM macrophage colonystimulating f...

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Effect of diclofenac on SLC16A3/MCT4 by the Caco-2 cell line

Cited by 42 publications

References 26 publications

Restricting Glycolysis Preserves T Cell Effector Functions and Augments Checkpoint Therapy

Restricting Glycolysis Preserves T Cell Effector Functions and Augments Checkpoint Therapy

Metabolic Hallmarks of Tumor and Immune Cells in the Tumor Microenvironment

MCT4 is a high affinity transporter capable of exporting lactate in high-lactate environments

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