Glycolysis Inhibition Sensitizes Non–Small Cell Lung Cancer with T790M Mutation to Irreversible EGFR Inhibitors via Translational Suppression of Mcl-1 by AMPK Activation

Abstract

The secondary EGF receptor (EGFR) T790M is the most common mechanism of resistance to reversible EGFR-tyrosine kinase inhibitors (TKI) in patients with non-small cell lung cancer (NSCLC) with activating EGFR mutations. Although afatinib (BIBW2992), a second-generation irreversible EGFR-TKI, was expected to overcome the acquired resistance, it showed limited efficacy in a recent phase III clinical study. In this study, we found that the inhibition of glycolysis using 2-deoxy-D-glucose (2DG) improves the efficacy of afatinib in H1975 and PC9-GR NSCLC cells with EGFR T790M. Treatment with the combination of 2DG and afatinib induced intracellular ATP depletion in both H1975 and PC9-GR cells, resulting in activation of AMP-activated protein kinase (AMPK). AMPK activation played a central role in the cytotoxicity of the combined treatment with 2DG and afatinib through the inhibition of mTOR. The alteration of the AMPK/mTOR signaling pathway by the inhibition of glucose metabolism induced specific downregulation of Mcl-1, a member of the antiapoptotic Bcl-2 family, through translational control. The enhancement of afatinib sensitivity by 2DG was confirmed in the in vivo PC9-GR xenograft model. In conclusion, this study examined whether the inhibition of glucose metabolism using 2DG enhances sensitivity to afatinib in NSCLC cells with EGFR T790M through the regulation of the AMPK/mTOR/Mcl-1 signaling pathway. These data suggest that the combined use of an inhibitor of glucose metabolism and afatinib is a potential therapeutic strategy for the treatment of patients with acquired resistance to reversible EGFR-TKIs due to secondary EGFR T790M.

Dual inhibition of Mcl-1 by the combination of carfilzomib and TG02 in multiple myeloma

“…Inhibition of Mcl-1 translation has been previously demonstrated as a means to induce cell death in tumor cells. [38][39][40] To verify that Mcl-1 is the key target in the combination treatment we created cells that over express the anti-apoptotic proteins. Overexpression of Mcl-1 in RPMI-8226 cells confers significantly less protection compared to over expression of Bcl-x L .…”

2-Deoxy-D-glucose targeting of glucose metabolism in cancer cells as a potential therapy

“…Decreased intracellular ATP production leads to an increase of the AMP/ATP ratio, which activates and increases the AMP-activated protein kinase (AMPK), resulting in elevated catabolic metabolism via phosphorylation of downstream targets, such as the mammalian target of rapamycin (mTOR) [42]. In addition, decreased ATP sensitizes cells to extrinsic (death receptor-mediated) apoptosis [44,45] via binding of the tumor necrosis factor (TNF) ligand family members, including TNF-related apoptosis-inducing ligand (TRAIL), to their cognate transmembrane death receptors [41].…”

“…Although 2DG is commonly thought to inhibit glycolysis, it actually has extensive metabolic effects and interferes with various biological processes, including depletion of cellular energy, intensification of oxidative stress, interference with N-linked glycosylation, and induction of autophagy [39,40], which also activates multiple signaling pathways, such as PI3K, MAPK, and AMPK [41,42]. These events are somewhat related and integrated.…”

A comprehensive review of the preclinical efficacy profile of the ErbB family blocker afatinib in cancer

“…This combination was reported to be synergistic for in vivo inhibition of lung cancer growth in a transgenic EGFR L858R/T790M model (Li et al 2008) and in transgenic as well as xenograft HER2 mutant models (Perera et al 2009). Inhibition of glycolysis using 2-deoxy- d -glucose sensitized NSCLC cells with EGFR T790M mutation to afatinib via translational suppression of Mcl-1 by activation of AMP-activated protein kinase (Kim et al 2013). This long list of examples demonstrates that afatinib provides ample opportunities for combinations with targeted agents.…”