Abstract 4162: T-cell metabolic profiling with bioluminescent assays

Abstract

Abstract Cellular energy metabolism is crucial for cell survival and the balance of metabolic pathways determines the health of the cell. Determining and understanding metabolic profiles of cells can aid in overall insight of cell health and improvement of cell therapeutics, especially for immune interventions. T-cells actively regulate their metabolic profile from quiescent to a highly metabolically active state after activation. Activation rewires the cell to drive glycolysis and oxidative phosphorylation for production of reducing power and ATP for cell growth and proliferation. A small population of these cells then transition to a more quiescent state and persist as memory cells to aid in secondary responses. Although the trend of T-cell metabolism after activation has been studied, the method of investigation is cumbersome and lacks depth of information of overall cell health. To address this, we developed a bioluminescence NAD(P)H core technology and applied it to build a set of cell-based assays covering diverse metabolic pathways including glucose, glycogen, and amino acid metabolism, and the TCA cycle. Here we utilize those assays to investigate changes in metabolic profiles of naïve, effector, and memory T-cells as they transition from a state of oxidative phosphorylation to increased glycolytic rates, dependence on different fuel consumption and storage, including an increase in fatty acid oxidation and glycogen accumulation. We demonstrate flexibility of the assays for detecting multiple metabolites from the same sample and monitoring metabolite changes over time by assaying multiple samples. We show that the assays are amenable to various volumes (96-,384-well plates) and low cell numbers. For example, activity of dehydrogenases involved in glycolysis, pentose phosphate and TCA cycles can be detected with less than 400 cells/well. In conclusion, understanding the processes governing dynamic metabolic changes of T-cells during their transition from naïve to activated to memory phenotypes is required for next-generation cell therapy development. The developed bioluminescence cell-based assays provide the sensitivity, throughput and robustness required for rapid evaluation of such changes. Citation Format: Maggie Bach, Natasha Karassina, Julia Gilden, Donna Leippe, Mike Valley, Kayla Sylvester, Jolanta Vidugiriene. T-cell metabolic profiling with bioluminescent assays. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4162.