Optimisation of immunofluorescence methods to determine MCT1 and MCT4 expression in circulating tumour cells

Kershaw, Stephen; Cummings, Jeffrey L.; Morris, Karen; Tugwood, Jonathan

doi:10.1186/s12885-015-1382-y

Cited by 10 publications

(6 citation statements)

References 38 publications

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“…The expression levels of HK2 and MCT1 were significantly higher in the CTC fraction from patients than in the corresponding "PBMC" fraction from HD (p = 0.015 and p = 0.005, respectively).However, no statistically significant difference was observed for PHDGH. These results are in line with other studies that have used immunofluorescence assays [55]. It is important to mention that HK2 has recently been used as a metabolic-function-associated marker for the identification of CTCs from NSCLC patients and has revealed a HK2high/CKneg CTC population which is not accessible to, and is normally overlooked by, current epithelial-marker-based CTC detection methods [29].…”

Section: Discussionsupporting

confidence: 89%

“…By using novel immunofluorescence methods, Kershaw et al have shown the overexpression of MCT1 in CTCs enriched in the Veridex™ Cell Search system, demonstrating that the detection of MCT1 in CTCs could offer the potential to act as a sensitive and selective biomarker assay in the clinical evaluation of drugs targeting this protein [55]. In the current study and for the first time, we studied the expression of PHGDH, MCT1 and HK2 in CTC fractions isolated from early stage NSCLC patients using size-based EpCAM independent technology (Parsortix).…”

Section: Discussionmentioning

confidence: 99%

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

Zafeiriadou

Kollias

Londra

et al. 2022

Cancers

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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.

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Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

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

Zafeiriadou

Kollias

Londra

et al. 2022

Cancers

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“…Evidence on cell and animal models revealed that MCT1 inhibition in ECs can drive direct anti-angiogenic effects through the enhanced degradation of HIF-1α. Drugs targeting MCT1 in a non-selective manner are currently been tested in small-scale trials (Kershaw et al, 2015). On the other hand, administration of the telomerase activator TA-65 contributes to improve blood flow recovery through increasing expression of HIF-1α, VEGF-A, and peroxisome proliferator-activated receptor (PPAR)-γ coactivator 1-alpha (PGC-1α), indicating that telomerase activation could prove a valuable therapeutic option to rescue ischemic tissues in elderly individuals (Kokubun et al, 2019).…”

Section: Therapeutic Interventions Targeting Endothelial Cell Metabolismmentioning

confidence: 99%

Where Metabolism Meets Senescence: Focus on Endothelial Cells

et al. 2019

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Despite the decline in their proliferative potential, senescent cells display a high metabolic activity. Senescent cells have been shown to acquire a more glycolytic state even in presence of high oxygen levels, in a way similar to cancer cells. The diversion of pyruvate, the final product of glycolysis, away from oxidative phosphorylation results in an altered bioenergetic state and may occur as a response to the enhanced oxidative stress caused by the accumulation of dysfunctional mitochondria. This metabolic shift leads to increased AMP/ATP and ADP/ATP ratios, to the subsequent AMPK activation, and ultimately to p53-mediated growth arrest. Mounting evidences suggest that metabolic reprogramming is critical to direct considerable amounts of energy toward specific activities related to the senescent state, including the senescence-associated secretory phenotype (SASP) and the modulation of immune responses within senescent cell tissue microenvironment. Interestingly, despite the relative abundance of oxygen in the vascular compartment, healthy endothelial cells (ECs) produce most of their ATP content from the anaerobic conversion of glucose to lactate. Their high glycolytic rate further increases during senescence. Alterations in EC metabolism have been identified in age-related diseases (ARDs) associated with a dysfunctional vasculature, including atherosclerosis, type 2 diabetes and cardiovascular diseases. In particular, higher production of reactive oxygen species deriving from a variety of enzymatic sources, including uncoupled endothelial nitric oxide synthase and the electron transport chain, causes DNA damage and activates the NAD +-consuming enzymes polyADP-ribose polymerase 1 (PARP1). These non-physiological mechanisms drive the impairment of the glycolytic flux and the diversion of glycolytic intermediates into many pathological pathways. Of note, accumulation of senescent ECs has been reported in the context of ARDs. Through their pro-oxidant, pro-inflammatory, vasoconstrictor, and prothrombotic activities, they negatively impact on vascular physiology, promoting both the onset and development of ARDs. Here, we review the current knowledge on the cellular senescence-related metabolic changes and their contribution to the mechanisms underlying the pathogenesis of ARDs, with a particular focus on ECs. Moreover, current and potential interventions aimed at modulating EC metabolism, in order to prevent or delay ARD onset, will be discussed.

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“…Although the expression of MCT4 in the tumor microenvironment has been associated with poor prognosis ( Nakayama et al, 2012 ; Baek et al, 2014 ) and MCT4 has been studied as a new therapeutic target ( Kim et al, 2018 ; Puri and Juvale, 2020 ). Immunofluoresence has been used for the detection of MCT 1 and MCT4 in cancer patient’s CTC by Kershaw et al (2015) but so far there is no any other reference on MCT4 expression at the mRNA level on CTCs.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Monocarboxylate Transporter 4 (MCT4) Expression and Its Prognostic Significance in Circulating Tumor Cells From Patients With Early Stage Non-Small-Cell Lung Cancer

Markou

Tzanikou

Kallergi

et al. 2021

Front. Cell Dev. Biol.

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Purpose: Monocarboxylate transporter 4 (MCT4) can influence the amount of lactate in the tumor microenvironment and further control cancer cell proliferation, migration, and angiogenesis. We investigated for the first time the expression of MCT4 in circulating tumor cells (CTCs) derived from early stage Non-Small Cell Lung Cancer patients (NSCLC) and whether this is associated with clinical outcome.Experimental Design: A highly sensitive RT-qPCR assay for quantification of MCT4 transcripts was developed and validated and applied to study MCT4 expression in CTC isolated through the Parsortix size-dependent microfluidic device from 53 and 9 peripheral blood (PB) samples of NSCLC patients at baseline (pre-surgery) and at relapse, respectively, as well as the “background noise” was evaluated using peripheral blood samples from 10 healthy donors (HD) in exactly the same way as patients.Results:MCT4 was differentially expressed between HD and NSCLC patients. Overexpression of MCT4 was detected in 14/53 (26.4%) and 3/9 (33.3%) patients at baseline and at progression disease (PD), respectively. The expression levels of MCT4 was found to increase in CTCs at the time of relapse. Kaplan-Meier analysis showed that the overexpression of MCT4 was significantly (P = 0.045) associated with progression-free survival (median: 12.5 months, range 5–31 months).Conclusion:MCT4 overexpression was observed at a high frequency in CTCs from early NSCLC patients supporting its role in metastatic process. MCT4 investigated as clinically relevant tumor biomarker characterizing tumor aggressiveness and its potential value as target for cancer therapy. We are totally convinced that MCT4 overexpression in CTCs merits further evaluation as a non-invasive circulating tumor biomarker in a large and well-defined cohort of patients with NSCLC.

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Optimisation of immunofluorescence methods to determine MCT1 and MCT4 expression in circulating tumour cells

Cited by 10 publications

References 38 publications

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

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

Where Metabolism Meets Senescence: Focus on Endothelial Cells

Evaluation of Monocarboxylate Transporter 4 (MCT4) Expression and Its Prognostic Significance in Circulating Tumor Cells From Patients With Early Stage Non-Small-Cell Lung Cancer

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