Pharmacology and preclinical validation of a novel anticancer compound targeting PEPCK-M

Aragó, Marc; Moreno-Felici, Juan; Abás, Sònia; Rodríguez-Arévalo, Sergio; Hyroššová, Petra; Figueras, Agnés; Viñals, Francesc; Pérez, Belén; Loza, Marı́a Isabel; Brea, José; Latorre, Pedro; Carrodeguas, José A.; García-Rovés, Pablo M.; Galdeano, Carles; Ginex, Tiziana; Luque, Francisco; Escolano, Carmen; Perales, José C.

doi:10.1016/j.biopha.2019.109601

Cited by 10 publications

(14 citation statements)

References 24 publications

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“…NFAT and c-Myc produce activation of expression of several genes implicated in inflammation and metastasis (PTGS2), glucose uptake and glycolysis (Glut1 and HKII), glutamine catabolism (GLS1) and serine and glycine synthesis, and transference of carbons to one-carbon metabolism (cSHMT) that allow for NADPH recycling. This scenario indicates that glucose and PEPCK-M are supporting, through the PEP/Ca 2+ axis, a proliferative state in tumors, and bring about the importance of this target for cancer metabolism, as exemplified by the efficacy of potent inhibitors of this pathway in pre-clinical studies [22].…”

Section: Discussionmentioning

confidence: 96%

“…Cellular PEP levels are also maintained by the PEPCK-dependent cataplerosis of TCA cycle carbons that contribute to the synthesis of glycolytic intermediates (Hyrossova et al, unpublished), serine/glycine [19] and triglycerides [20,21]. In most tumor models, and specifically in the cellular models utilized in this work, the mitochondrial isoform of PEPCK (PEPCK-M) plays this key role as it is the only isoform of PEPCK expressed [22]. Therefore, we interrogated the implication of PEPCK-M in the regulation of PEP and Ca 2+ fluctuations, we measured PEP and Ca 2+ at various levels of activation of PEPCK-M more specifically using a potent inhibitor of PEPCK-M prepared in our laboratory, iPEPCK-2 [22], in the presence or absence of glucose ( Figure 1G).…”

Section: Glycolytic and Cataplerotic Fluxes Towards Pep Cooperate To mentioning

confidence: 99%

“…In most tumor models, and specifically in the cellular models utilized in this work, the mitochondrial isoform of PEPCK (PEPCK-M) plays this key role as it is the only isoform of PEPCK expressed [22]. Therefore, we interrogated the implication of PEPCK-M in the regulation of PEP and Ca 2+ fluctuations, we measured PEP and Ca 2+ at various levels of activation of PEPCK-M more specifically using a potent inhibitor of PEPCK-M prepared in our laboratory, iPEPCK-2 [22], in the presence or absence of glucose ( Figure 1G). PEPCK-M activity contributed to the PEP/Ca 2+ axis in both colon carcinoma cell lines in the presence of glucose, as its inhibition effectively reduced cellular PEP concentrations and cytosolic calcium ( Figure 1H,I).…”

Section: Glycolytic and Cataplerotic Fluxes Towards Pep Cooperate To mentioning

confidence: 99%

“…In addition, we have demonstrated that changes in Ca 2+ produced by different glucose and PEP concentrations lead to activation and nuclear translocation of NFAT. For these reasons, we analyzed PEPCK-M contribution to NFAT activation by measuring NFAT activity and NFAT cellular localization in the presence or absence of a potent PEPCK-M inhibitor previously validated in our laboratory (iPEPCK-2; [22]; Figure 4). A reduction in NFAT activity was observed when PEPCK-M was inhibited in the presence of glucose in SW480 cells ( Figure 4A), consistent with reduced NFAT activity found in SW480 and HCT116 cell lines upon glucose depletion.…”

Section: Pepck-m Modulates Calcium Signaling Because Of Its Impact Onmentioning

confidence: 99%

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Phosphoenolpyruvate from Glycolysis and PEPCK Regulate Cancer Cell Fate by Altering Cytosolic Ca2+

Moreno-Felici

Hyroššová

Aragó

et al. 2019

Cells

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Changes in phosphoenolpyruvate (PEP) concentrations secondary to variations in glucose availability can regulate calcium signaling in T cells as this metabolite potently inhibits the sarcoplasmic reticulum Ca 2+ /ATPase pump (SERCA). This regulation is critical to assert immune activation in the tumor as T cells and cancer cells compete for available nutrients. We examined here whether cytosolic calcium and the activation of downstream effector pathways important for tumor biology are influenced by the presence of glucose and/or cataplerosis through the phosphoenolpyruvate carboxykinase (PEPCK) pathway, as both are hypothesized to feed the PEP pool. Our data demonstrate that cellular PEP parallels extracellular glucose in two human colon carcinoma cell lines, HCT-116 and SW480. PEP correlated with cytosolic calcium and NFAT activity, together with transcriptional up-regulation of canonical targets PTGS2 and IL6 that was fully prevented by CsA pre-treatment. Similarly, loading the metabolite directly into the cell increased cytosolic calcium and NFAT activity. PEP-stirred cytosolic calcium was also responsible for the calmodulin (CaM) dependent phosphorylation of c-Myc at Ser62, resulting in increased activity, probably through enhanced stabilization of the protein. Protein expression of several c-Myc targets also correlated with PEP levels. Finally, the participation of PEPCK in this axis was interrogated as it should directly contribute to PEP through cataplerosis from TCA cycle intermediates, especially in glucose starvation conditions. Inhibition of PEPCK activity showed the expected regulation of PEP and calcium levels and consequential downstream modulation of NFAT and c-Myc activities. Collectively, these results suggest that glucose and PEPCK can regulate NFAT and c-Myc activities through their influence on the PEP/Ca 2+ axis, advancing a role for PEP as a second messenger communicating metabolism, calcium cell signaling, and tumor biology.

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Section: Discussionmentioning

confidence: 96%

Section: Glycolytic and Cataplerotic Fluxes Towards Pep Cooperate To mentioning

confidence: 99%

Section: Glycolytic and Cataplerotic Fluxes Towards Pep Cooperate To mentioning

confidence: 99%

Section: Pepck-m Modulates Calcium Signaling Because Of Its Impact Onmentioning

confidence: 99%

See 2 more Smart Citations

Phosphoenolpyruvate from Glycolysis and PEPCK Regulate Cancer Cell Fate by Altering Cytosolic Ca2+

Moreno-Felici

Hyroššová

Aragó

et al. 2019

Cells

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show abstract

“…Therefore, PEPCK acts as a mediator of cancer cell metabolic adaptation, allowing for glucose-independent tumor growth and proliferation [8]. Of note, PEPCK inhibition or silencing reduced the growth of tumors in different in vivo models [11][12][13]20]. On the other hand, growth inhibitory effects of PEPCK have been found in liver and kidney cancer cells (for review, see [8,21]).…”

Section: Introductionmentioning

confidence: 99%