Abstract 441: Bioluminescent dehydrogenase assays: A novel approach for metabolic profiling and inhibitor screening in cancer biology

Abstract

Abstract Dehydrogenases play a pivotal role in cellular metabolism, acting as gatekeepers in key metabolic pathways. Their activity reflects the metabolic state of the cell, which is particularly relevant in the context of cancer and the immune response, where metabolic reprogramming is a hallmark of tumor progression and differentiation stages. Here, we present the development and application of novel bioluminescent dehydrogenase assays for direct measurement of metabolic activity in cell lysates. Our assays are highly sensitive, requiring as few as 100-500 cells, and have been successfully miniaturized to a 384-well plate format conducive to high-throughput screening. This technological advance allows for the precise monitoring of dehydrogenase activity across the pentose phosphate pathway, glycolysis, and the mitochondrial tricarboxylic acid (TCA) cycle. We demonstrate how shifts in dehydrogenase activity can serve as indicators of the metabolic status of cells, and their modulation during differentiation. Moreover, our bioluminescent assays provide a rapid screening platform for pathway-specific metabolic inhibitors, with potential applications in identifying new therapeutic targets within cancer metabolism. Our data reveal a dynamic landscape of metabolic activity during T cell maturation, underscoring the potential of dehydrogenase activity profiling in cancer immunobiology and the development of targeted cancer therapies. Citation Format: Kim Haupt, Natasha Karassina, Anthony Lauer, Kayla Sylvester, Jolanta Vidugiriene. Bioluminescent dehydrogenase assays: A novel approach for metabolic profiling and inhibitor screening in cancer biology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 441.