Real-time metabolic profiling of oesophageal tumours reveals an altered metabolic phenotype to different oxygen tensions and to treatment with Pyrazinib

Abstract

0.1% DMSO vehicle control or 0, 0.1, 1, 5 or 10 µM of pyrazinib (P3), after which time they were transferred to a FACS tube and washed in 1 mL 1X binding buffer. Cells were stained with 3 μl of Annexin V antibody and incubated in the dark for 15-20 min at 4 °C. Cells were washed and resuspended in 250 μL of 1 in 4,000 dilution of PI in 1X binding buffer and samples were immediately acquired on FACSCanto II flow cytometer (BD Biosciences). Assessing the activation status of Jurkat cells. Unactivated and activated Jurkat cells were seeded 5 × 10 4 cells/mL in 12 well plates in complete RPMI. Jurkats were activated with plate bound anti-CD3 (Biolegend, CA, USA) and anti-CD28 (Ancell, MN, USA) monoclonal antibodies for 24 h. Cells were treated for 24 or 48 h with 0.1% DMSO vehicle control or 10 µM of pyrazinib (P3). Cells were washed and blocked and then stained with anti-CD62 PerCp-Cy5 (Abcam, UK), anti-CD69 PE (BD Biosciences, CA, USA), anti-CD45RO FITC (Immunotools, Germany), anti-CD45RA V500 (BD Biosciences) and anti-CD27 APC eFluor780 (eBioscience. CA, USA) for 20 min at 4 °C in the dark. Cells were washed and resuspended in 300 μL FACS buffer and acquired on a FACSCanto II flow cytometer (BD Biosciences). Statistical analysis. Statistical analysis was performed using GraphPad Prism software version 5 (Graph-Pad Software, CA, USA). Scientific data were expressed as mean ± SEM. Specific statistical tests used are indicated in figure legends. For all statistical analysis, differences were considered to be statistically significant at p < 0.05.

Energy Metabolism Is Altered in Radioresistant Rectal Cancer

“…Interestingly, treatment with the potent glycolysis inhibitor 2-DG significantly sensitized HCT116 cells to 1.8 Gy radiation ( Figure 5 G), an effect not demonstrated in SW837 cells, further supporting a decreased dependence on glycolysis in these radioresistant cells and suggesting that this is important for the radioresponse of these cells. Importantly, these findings support previous data from our lab, highlighting the importance of oxidative phosphorylation in the radioresistance of esophageal adenocarcinoma [ 34 , 35 , 36 ] and suggest a common role for metabolic reprogramingin the response of gastrointestinal cancers to radiation. Importantly, cell cycle and DNA repair, which were demonstrated to be altered in radioresistant SW837 cells, require cellular energy availability and are subsequently functionally dependent on energy metabolism, highlighting the potential central role for metabolic pathways in determining tumoral radioresponse in rectal cancer.…”

Potential Predictive Immune and Metabolic Biomarkers of Tumor Microenvironment Regarding Pathological and Clinical Response in Esophageal Cancer After Neoadjuvant Chemoradiotherapy: A Systematic Review

“…In clinical imaging of hypoxic response and glycolytic metabolism in malignant tumors, 18 F-FDG-PET/CT is most commonly used. 50 Based on the assessment of histopathology, the corresponding 18 F-FDG-PET/CT response and promising biomarkers markers, nCRT combined with immunotherapy might be considered as an organ-preserving treatment approach in the near future.…”

Energy Metabolism, Metabolite, and Inflammatory Profiles in Human Ex Vivo Adipose Tissue Are Influenced by Obesity Status, Metabolic Dysfunction, and Treatment Regimes in Patients with Oesophageal Adenocarcinoma

“…Previous work has observed elevated oxidative phosphorylation in visceral adipose tissue compared with subcutaneous adipose tissue [ 16 ], and adipocytes derived from metabolically unhealthy obese individuals show elevated mitochondrial response profiles [ 19 ]. Whilst the role of tumour explant energy metabolism in OAC treatment response has been reported [ 20 ], the energy metabolism profiles of visceral adipose tissue between obese and non-obese OAC patients and the influence of its secretome on the cancer cell and immune cell function is still largely unknown.…”