(175s) Comparison of Metabolic Gene Expression Levels between Lactate-Consuming and Lactate-Producing CHO Cell Cultures
AIChE Annual Meeting
2019
2019 AIChE Annual Meeting
Food, Pharmaceutical & Bioengineering Division
Poster Session: Bioengineering
Monday, November 11, 2019 - 3:30pm to 5:00pm
Glucose metabolism is a vital process that cells use to produce energy via glycolysis by breaking down pyruvate, either reductively metabolized to lactate (fermentation), or oxidatively phosphorylated to produce ATP via the TCA cycle. It has been shown that while the former pathway is less efficient in energy production, and despite the ample availability of oxygen, highly proliferating cells show a marked preference toward energy production by the reductive metabolization of pyruvate to lactate in what is known as the Warburg effect. An important aspect of the glycolytic flux is the need to regenerate NAD+. This can be accomplished either by lactate fermentation or via the malate-aspartate shuttle (MAS) across the mitochondrion. It has been demonstrated that MAS activity is crucial in determining the ratio of pyruvate oxidized in the TCA cycle versus pyruvate converted to lactate. The scope of this work was to use One-step RT qPCR to analyze the genetic expression levels of various genes associated with glycolysis and fermentation pathways, such as HK1, HIF-1α, PFKM, and LDHA, as well as those involved in the MAS system, such as Timm13 and Aralar1, in cell cultures that exhibited either a lactate-accumulating, or a lactate-consuming phenotype. The analysis showed that genes that support the glycolysis pathway such as HIF-1α, G6PD, and HK1 were upregulated in cultures that experienced base addition and displayed subsequent lactate-accumulating behavior, while genes associated with pathways leading to lower glycolytic flux, such as Timm13 and Aralar1, showed downregulation.