Co-occurring KRAS mutation/LKB1 loss in non-small cell lung cancer cells results in enhanced metabolic activity susceptible to caloric restriction: an in vitro integrated multilevel approach

Abstract

BackgroundNon–small-cell lung cancer (NSCLC) is a heterogeneous disease, with multiple different oncogenic mutations. Approximately 25–30% of NSCLC patients present KRAS mutations, which confer poor prognosis and high risk of tumor recurrence. About half of NSCLCs with activating KRAS lesions also have deletions or inactivating mutations in the serine/threonine kinase 11 (LKB1) gene. Loss of LKB1 on a KRAS-mutant background may represent a significant source of heterogeneity contributing to poor response to therapy.MethodsHere, we employed an integrated multilevel proteomics, metabolomics and functional in-vitro approach in NSCLC H1299 isogenic cells to define their metabolic state associated with the presence of different genetic background. Protein levels were obtained by label free and single reaction monitoring (SRM)-based proteomics. The metabolic state was studied coupling targeted and untargeted mass spectrometry (MS) strategy. In vitro metabolic dependencies were evaluated using 2-deoxy glucose (2-DG) treatment or glucose/glutamine nutrient limitation.ResultsHere we demonstrate that co-occurring KRAS mutation/LKB1 loss in NSCLC cells allowed efficient exploitation of glycolysis and oxidative phosphorylation, when compared to cells with each single oncologic genotype. The enhanced metabolic activity rendered the viability of cells with both genetic lesions susceptible towards nutrient limitation.ConclusionsCo-occurrence of KRAS mutation and LKB1 loss in NSCLC cells induced an enhanced metabolic activity mirrored by a growth rate vulnerability under limited nutrient conditions relative to cells with the single oncogenetic lesions. Our results hint at the possibility that energy stress induced by calorie restriction regimens may sensitize NSCLCs with these co-occurring lesions to cytotoxic chemotherapy.Electronic supplementary materialThe online version of this article (10.1186/s13046-018-0954-5) contains supplementary material, which is available to authorized users.

Searching for Drug Synergy Against Cancer Through Polyamine Metabolism Impairment: Insight Into the Metabolic Effect of Indomethacin on Lung Cancer Cells

“…do not overexpress carbamoyl phosphate synthetase-1, without induction of the urea cycle (Celiktas et al, 2017). However, in a model of H1299 cells-mutated KRAS, the urea cycle and polyamine metabolism are upregulated with a significant increase in the levels of putrescine and spermidine (Caiola et al, 2018).…”

Glutaminase Inhibition on NSCLC Depends on Extracellular Alanine Exploitation

“…As previously described in [ 9 ], 13C 5 -labelled-glutamine analysis was carried out on NSCLC H1299 derived cell clones. Briefly, NSCLC cell clones (1 × 10 6 cells) were cultured for 24 h in RPMI -1640 media supplemented with 10% dialyzed FBS and 1% standard glutamine; then, cells were treated with CB-839 (500 nM), and after 18 h, medium was replenished with RPMI with 2 mM 13C 5 -glutamine (Cambridge Isotopes Laboratories, Tewksbury, MA, USA) and CB-839 (500 nM) for 6 h. To trace alanine metabolism, NSCLC H1299-R1 and -R2 derived cell clones, and LU99, A549, H358 and H520 cell lines were seeded at the desired concentrations and, after 24 h, medium was supplemented with 1 mM U-13C 3 -alanine and CB-839 (500 nM) for 24 h. After labelling and treatment, conditioned culture media were collected, cells were rinsed, and metabolism was quenched with liquid nitrogen.…”

Regulation of metabolic reprogramming by tumor suppressor genes in pancreatic cancer

“…The interactions between different genes promote the initiation and development of tumors, but at the same time provide a new strategy for us to the treatment of cancer. For example, Caiola et al reported that co-occurring mutation of KRAS and LKB1 in NSCLC cells showed more efficient glycolysis and oxidative phosphorylation compared to cells with either single mutation genotype, however the enhanced metabolic activity renders cells with both genetic lesions more sensitive to nutrient limitation, suggesting the possibility to kill cancer cells through energy stress which induced by nutrition restriction regimens [97].…”

“… [97]. Preventing increased mutation rate and karyotype instability caused by p53 downregulation caused excessive oxidation of DNA Experimental and/or preclinical Lymphoma and Lung cancer 2005FX11Rajeshkumar et al[26] Inhibiting lactate dehydrogenase-A (LDH-A) which promote pyruvate to metabolize to lactate during the process of aerobic glycolysisExperimental and/or preclinical Pancreatic cancer 2015 APR-246 Peng et al [99] Inhibits the oxidoreductase enzyme thioredoxin reductase 1 (TRXR1) and converts the enzyme to a pro-oxidant NADPH oxidase, thereby inducing oxidative stress Experimental and/or preclinical H1299-His175, glutathione (GSH) and thereby inducing lipid peroxidative cell death Experimental and/or preclinical Oesophageal cancer 2017 APR-246 Ali et al [102] Increasing expression of genes that are related to oxidative stress including haeme oxygenase 1 (HMOX1) and so on Experimental and/or preclinical Acute myeloid leukaemia 2016 APR-246 Lambert et al [100] Inducing endoplasmic reticulum stress by its redox effects Experimental and/or preclinical Saos-2 cells 2010 Pramlintide Venkatanarayan et al [98] Inhibiting glycolysis and inducing reactive oxygen species (ROS) and apoptosis through calcitonin receptor (CalcR) and receptor activity modifying protein 3 (RAMP3) Experimental and/or preclinical thymic lymphoma 2015 LKB1 ND-646 Svensson et al [119] Inhibiting Acetyl-CoA carboxylase (ACC), which is a product in the first step of fatty acid (FA) synthesis, thus inhibiting FA synthesis and tumor growth Experimental and/or preclinical Non-small-cell lung cancer 2016 Adenoviruses expressing Cre and/or Flp recombinase Kottakis et al [117] Induction of the serine-glycine-one-carbon pathway coupled to S-adenosylmethionine generation, and thus sensitizes cells and tumors to inhibition of serine biosynthesis Experimental and/or preclinical Pancreatic cancer 2016 Cb-839 Galan-Cobo et al [86] Enhanced energetic/redox stress, tolerated through activation of KEAP1/NRF2-in a glutamine-dependent manner enhanced glutamine dependence and vulnerability to glutaminase inhibition Experimental and/or preclinical Non-small-cell lung cancer 2019 PTEN Avasimibe Li et al [123] ACAT inhibition increased intracellular free cholesterol level and caused elevated endoplasmic reticulum stress and apoptosis Experimental and/or preclinical Pancreatic cancer 2016 Oroxylin A Zhao et al [103] Inducing the downregulation of mouse double minute 2 (MDM2) transcription by promoting the lipid phosphatase activity of PTEN, and further suppress the MDM2-mediated degradation of p53, thereby inhibiting glycolysis Experimental and/or preclinical MCF-7 and HCT116 cells Page 12 of 16 Liu et al Exp Hematol Oncol (2020) 9:23…”