Metabolism-Related Gene Expression in Circulating Tumor Cells from Patients with Early Stage Non-Small Cell Lung Cancer

Abstract

Purpose: Metabolic reprogramming is now characterized as one of the core hallmarks of cancer, and it has already been shown that the altered genomic profile of metabolically rewired cancer cells can give valuable information. In this study, we quantified three Metabolism-Related Gene (MRG) transcripts in the circulating tumor cells (CTCs) of early stage NSCLC patients and evaluated their associations with epithelial and EMT markers. Experimental Design: We first developed and analytically validated highly sensitive RT-qPCR assays for the quantification of HK2, MCT1 and PHGDH transcripts, and further studied the expression of MRGs in CTCs that were isolated using a size-dependent microfluidic device (Parsortix, Angle) from the peripheral blood of: (a) 46 NSCLC patients at baseline, (b) 39/46 of these patients one month after surgery, (c) 10/46 patients at relapse and (d) 10 pairs of cancerous and adjacent non-cancerous FFPE tissues from the same NSCLC patients. Epithelial and EMT markers were also evaluated. Results: MCT1 and HK2 were differentially expressed between HD and NSCLC patients. An overexpression of MCT1 was detected in 15/46 (32.6%) and 3/10 (30%) patients at baseline and at progression disease (PD), respectively, whereas an overexpression of HK2 was detected in 30.4% and 0% of CTCs in the same group of samples. The expression levels of all tested MRGs decreased in CTCs one month after surgery, but a significant increase was noticed at the time of relapse for PHGDH and MCT1only. The expression levels of HK2 and MCT1 were associated with the overexpression of mesenchymal markers (TWIST-1 and VIM). Conclusion: An overexpression of MRGs was observed at a high frequency in the CTCs isolated from early NSCLC patients, thereby supporting the role of MRGs in metastatic processes. The glycolytic and mesenchymal subpopulation of CTCs was significantly predominant compared to CTCs that were glycolytic but not mesenchymal-like. Our data indicate that MRGs merit further evaluation through large and well-defined cohort studies.

A workflow for the enrichment, the identification, and the isolation of non‐apoptotic single circulating tumor cells for RNA sequencing analysis

“…The metabolic activity of CTCs has been investigated in several studies and high metabolic activity was associated with poor clinical outcome for colorectal cancer patients [30] and advanced tumor stage and metastasis in prostate cancer [31]. On the other hand, these studies also show that a large proportion of CTCs harbor no or reduced signs of metabolic activity: Chen et al did not detect markers for glucose metabolism in 53.5% of prostate cancer CTCs [31]; Zafeiriadou et al observed an overexpression of the metabolic markers monocarboxylate‐transporter 1, hexokinase 2, or phosphoglycerate dehydrogenase in only 30% or less of non‐small cell lung cancer CTCs [32]. Of note, in our study, the presence of transcripts for certain housekeeping genes such as the metabolism related gene GAPDH could clearly separate cells based on the quality of the RNA sequencing data supporting the effect of metabolism on RNA quality of CTCs.…”

The Nitric Oxide Donor [Zn(PipNONO)Cl] Exhibits Antitumor Activity through Inhibition of Epithelial and Endothelial Mesenchymal Transitions

“…In particular, non-small cell lung carcinoma (NSCL) and malignant melanoma share poor prognosis, and the therapeutic options are few and characterized by scarce effectiveness and resistance [ 39 , 40 ]. These malignancies show a predisposition to EMT and EndMT [ 41 , 42 , 43 , 44 ]. Few are the reports on the development of NO donors in these tumor types [ 18 , 45 , 46 , 47 , 48 ]; thus, the field merits investigation.…”