Abstract 3487: Bioenergetic profiling of cancer cell lines: Quantifying the impact of glycolysis on cell proliferation

Abstract

Fast proliferating cells require tight regulation to achieve a balance between the use of nutrients for ATP production (through glycolysis and oxidative phosphorylation) and the use of intermediate metabolites to sustain the increased biosynthetic activity. Cancer cells, but also high proliferative non-transformed cells exhibit high glycolytic activity during rapid proliferation even in the presence of normal oxygen concentrations in culture. However, despite the high glycolytic activity, the role of glycolysis is not necessary as a major contributor of ATP but to allow nutrient assimilation into biosynthetic precursors. Using Agilent Seahorse extracellular flux analysis, we have developed a cell-based assay which allows simultaneous measurement of the two-main cellular metabolic pathways to calculate the total rate of cellular ATP production as well as the fractional contribution from each pathway. The assay allows for real time changes in total ATP production rate to be quantified, and also the relative source of that ATP after exposure to drugs or changes in extracellular fuel supply. When we applied this new assay to a panel of 20 cancer and highly proliferative cell lines, we found that even in cell lines considered highly glycolytic, ATP production from glycolysis never represents more than 65% of total energy production and between 30-50% for most of the cell lines analyzed. The correlation between glycolytic ATP contribution to total ATP production and other cell phenotypes such as proliferation rate and motility was also analyzed. The use of this assay will allow for improved characterization of the bioenergetic profile of cancer cell variants, discrimination between normal and cancer cell types, and allow researchers to better understand the role of aerobic glycolysis in cell proliferation. Citation Format: Natalia Romero, Pamela M. Swain, Yoonseok Kam, George Rogers, Brian P. Dranka. Bioenergetic profiling of cancer cell lines: Quantifying the impact of glycolysis on cell proliferation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3487.

Disarib, a Specific BCL2 Inhibitor, Induces Apoptosis in Triple-Negative Breast Cancer Cells and Impedes Tumour Progression in Xenografts by Altering Mitochondria-Associated Processes

“…While acknowledging potential inherent gene expression variations in these lines, our primary focus was on Disarib’s impact on cellular energy metabolism. By leveraging published data on the metabolic profiles of both MDA-MB-231 and T47D cells, we were able to analyse and interpret the observed changes in a more targeted manner [ 26 , 27 ]. A separate heatmap was plotted for the genes belonging to OXPHOS, glycolysis, fission and fusion, and mitophagy pathways.…”

Depletion of pyruvate kinase (PK) activity causes glycolytic intermediate imbalances and reveals a PK-TXNIP regulatory axis

“…However, how depletion of a key rate-limiting enzyme in glycolysis, PK ( Figure 1 A) impacts on both transcriptional and epigenetic pathways is currently unclear. To determine transcriptional and histone modification changes upon induced depletion of PK activity in highly glycolytic cells, we established a stable PKM knockdown in human embryonic kidney 293 (HEK293) and in Michigan Cancer Foundation-7 (MCF-7) cell lines using shRNA targeting a common fragment of the spliced PKM1/2 transcript [ 64 , 65 ]. We confirmed the efficiency of the knockdowns of both isoforms at the level of mRNA and protein by RT-qPCR, RNA-seq and immunoblotting ( Figure 1 B–C).…”

Metabolic symbiosis between oxygenated and hypoxic tumour cells: An agent-based modelling study

“…where, A 0 is the ATP yield at the relevant oxygen and glucose abundance conditions, which is about 28 to 36 ATP molecules. The proportion of ATP production by glycolysis and OXPHOS depends on the cell type [36] and therefore, we assume that glycolytic and OXPHOS cells have similar ATP production rates for their proper functioning. The glucose consumption rate is then modelled as…”