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Sensitive Bioluminescent Assays for Monitoring Changes in Tumor and Immune Cell Metabolism

Part # PS338

Abstract

Cells of the tumor microenvironment must compete for fuels and respond to changing, dysregulated conditions to remain viable and functional. Identifying the metabolic requirements and vulnerabilities of tumor and immune cells in this environment can aid in the development of more effective cancer treatments.

We have developed a series of bioluminescent metabolite detection assays for monitoring key cell metabolic pathways such as: glycolysis by glucose and lactate detection; glutaminolysis with glutamine and glutamate measurements; and lipolysis and lipogenesis through glycerol and triglyceride measurements. The assays share a common NAD(P)H bioluminescent detection technology and were developed to have high sensitivity, an important feature when samples are limiting and contain only a few cells or small volumes.

We have used these assays to study the metabolic activity of tumor cell lines and T cells. Here we present an example of studying T cell activation in vitro, and its fuel requirements, by monitoring a switch to glycolysis and an increase in lactate secretion over time. We also used the assays to follow metabolic pathway alterations induced in cancer cell lines by treatment with small molecules. Treatment with such compounds might provide a means of manipulating tumor nutrient usage and metabolite secretion in the tumor microenvironment.

Printed in USA.